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Application │RTG 2099/1 Cancer Cell Dissemination Primary Resistance Novel Targets Heidelberg University DKFZ Heidelberg Designated Speaker: Prof. Dr. S. Goerdt Designated Vice-Speaker: Prof. Dr. M. Leverkus London Coordinator: Prof. A. Hayday, PhD Proposed Funding Period 01/04/2015 – 30/09/2019 Table of Contents Page 1 General Information 1 1.1 Title in English and German 1 1.2 Host University 1 1.3 Participating Researchers 1 1.4 Associated Researchers / Lecturers 2 1.5 Summary in English and German 2 1.5.1 Summary in English 2 1.5.2 Summary in German 3 1.6 Funding Period 4 1.7 Number of PhD and MD positions 4 2 Profile of the Research Training Group 4 3 Research Program 5 3.1 Overview and Aims 5 3.1.1 Research Area A – Cancer Cell Dissemination 6 3.1.2 Research Area B – Primary Resistance to Cell Death and Immunity 6 3.1.3 Novel Targets – A Cross-sectional Approach 7 3.1.4 Collaboration, Methods, Model Systems 7 3.2 Project Descriptions Research Area A – Cancer Cell Dissemination Project Package A1 Cancer Stem Cells 8 Project 1 (Sleeman) 8 Project 2 (Utikal) 10 Project 3 (I. Augustin/Boutros) 12 Project 4 (Boutros) 14 Project Package A2 Invasion and Metastasis 16 Project 5 (Angel) 16 Project 6 (Schneider/Winkler) 18 Project 7 (Géraud/Goerdt) 20 I Page Research Area B – Primary Resistance to Cell Death and Immunity Project Package B1 Primary Resistance to Cell Death 22 Project 8 (Felcht/H. Augustin) 22 Project 9 (Leverkus) 24 Project 10 (Geserick/Leverkus) 26 Project Package B2: Primary Resistance to Tumor Immunity 28 Project 11 (Lonsdorf/Enk) 28 Project 12 (Schäkel/Cerwenka) 30 Project 13 (Schmieder/Umansky) 32 4 Qualification Program 34 4.1 Qualification Program 34 4.1.1 Seminars 35 4.1.2 Laboratory Instruction 35 4.1.3 Workshops 37 4.1.4 Student Project Development Platforms and Students’ Conferences 37 4.1.5 Transition from the First Class of Students to the Next Class 37 4.2 Guest Scientist Program 37 4.3 Additional Qualification Program – Scientific Collaboration with the London Project Partners of the RTG 37 4.4 Only Regarding International Research Training Groups: Research Stays at the Partner Institution 38 5 Supervision and Career Development, Equal Opportunity / Gender Equality, Organization, and Quality Management 39 5.1 Application and Selection Concept 39 5.2 Supervising Concept and Career Development 39 5.3 Equal Opportunity / Gender Equality in Science 40 5.4 Organization 42 5.5 Additional Aspects of Quality Management 43 II Page 6 Scientific Environment 44 6.1 Demarcation from Existing SFBs 45 6.2 Demarcation from Preexisting Graduate Colleges 45 7 Modules / Requested Funding 45 7.1 Module Research Training Group 45 7.2 Module Substitute 47 7.3 Module Coordination 47 7.4 Module Rotational Positions 47 7.5 Module Mercator Fellows 47 7.6 Module Project-Specific Workshops 47 7.7 Module Public Relations 47 7.8 Module Start-up Grants 48 7.9 Module Equal Opportunity / Gender Equality 48 Table 1 48 Table 2 48 Table 3 48 8 Only Regarding International Research Training Groups: Complementary Funding by the Partner Institution 49 9 Declarations 49 9.1 Relations to other SFBs 49 9.2 Collaboration with other Cooperation Partners 49 9.3 Cooperation with Corporate Partners 49 9.4 Admission of Qualification Students 49 9.5 Submissions of the Proposal to other Funding Organizations 49 9.6 Only Regarding International Research Training Groups: Letter of Intent of the Partner Institution 49 10 Obligations 50 11 Signatures 50 III Appendix I 1. List of Published Research Relevant to the 51 Research Program Appendix II 1. Biographical Sketches of the Participating 70 Researchers 2. Biographical Sketches of the Associated Researchers Appendix III Declarations regarding Section 9.2. “Collaboration with other Cooperation Partners” 1. Letter of Intent by the Medical Faculty Heidelberg regarding funding 2. Letter of Intent by the Medical Faculty Mannheim regarding funding 3. Letter of Intent of the London Co-ordinator, Prof. A. Hayday, King’s College and Cancer Research UK, London, regarding Scientific Collaboration IV 113 124 1 General information 1.1 Title in English and German Hallmarks of Skin Cancer: Cancer Cell Dissemination, Primary Resistance, Novel Targets Mechanismen des Hautkrebses: Metastasierung, primäre Resistenz und neue Zielstrukturen 1.2 Host University Ruprecht-Karls-University Heidelberg and German Cancer Research Center 1.3 Participating Researchers JECT NAME, ACADEMIC TITLE, DATE OF BIRTH 1 DEPARTMENT, AFFILIATION , POSTAL ADDRESS 1 Prof. Dr. Jonathan Sleeman, 24.02.1965 Dept. Microvascular Biology and Pathobiology, MFM, Ludolf-KrehlStr. 13-17, 68167 Mannheim 2 Prof. Dr. Jochen Utikal, 09.11.1974 CCU Dermato-Oncology, DKFZ/MFM, UMM, 68135 Mannheim 3 Dr. Iris Augustin, 18.03.1969 Prof. Dr. Michael Boutros, 26.10.1970 4 Prof. Dr. Michael Boutros, 26.10.1970 5 Prof. Dr. Peter Angel, 24.02.1959 6 7 8 9 10 11 Prof. Dr. Stefan Schneider 10.11.1966 Prof. Dr. Frank Winkler 15.08.1971 PD Dr. Cyrill Géraud 01.06.1982 Prof. Dr. Sergij Goerdt 14.05.1959 Dr. Moritz Felcht 01.08.1979 Prof. Dr. Hellmut Augustin 05.02.1959 Prof. Dr. Martin Leverkus 27.12.1965 Dr. Peter Geserick 11.09.1976 Prof. Dr. Martin Leverkus 27.12.1965 Dr. Anke Lonsdorf 08.02.1977 Prof. Dr. Alexander Enk 10.05.1963 Division of Signaling and Functional Genomics, DKFZ, and Dept. Cell and Molecular Biology, MFM, INH 280, 69120 Heidelberg Division of Signaling and Functional Genomics, DKFZ, and Dept. Cell and Molecular Biology, MFM, INH 280, 69120 Heidelberg CONTACT (FON, FAX, EMAIL) Fon: 0621-383-9965 Fax: 0621-383-9961 Jonathan.sleeman@med ma.uni-heidelberg.de DermatoOncology, Melanoma, Stem Cells Fon: 06221-421955 Fax: 06221-421959 [email protected] Signaling and Functional Genomics Fon: 06221-42-1951 Fax: 06221-42-1959 [email protected] Signaling and Functional Genomics Fon: 06221-42-4570 Fax: [email protected] Dept. Experimental Dermatology, MFM; UMM, 68135 Mannheim Dept. Neuro-Oncology, MFH, INH 400, 69120 Heidelberg Fon: 0621-338-6901 Fax: 0621-383-6903 stefan.schneider@medm a.uni-heidelberg.de Fon: 0621-383-2280 Fax: 0621-383-3815 [email protected] [email protected] Fon: 0621-383-2280 Fax: 0621-383-3815 [email protected] [email protected] Fon: 0621-383-2344 Fax: 0621-383-4085 Martin.Leverkus@medm a.uni-heidelberg.de Fon: 0621-383-2344 Fax: 0621-383-4085 Peter.Geserick@medma. uni-heidelberg.de Fon: 06221-56-8500 Fax: 06221-56-5406 [email protected] Fon: 06221.56-8447 Fax: 06221-56-5406 [email protected] [email protected] Fon: 0621-383-2048 Fax: 0621-383-3815 Astrid.schmieder@umm. de Dept. Dermatology, MFM; Vascular Oncology, MFM and DKFZ; UMM, 68135 Mannheim Dept. Molecular Dermatology, MFM, UMM, 68135 Mannheim Dept. Molecular Dermatology, MFM, UMM, 68135 Mannheim Dept. Dermatology, MFH, Voßstr.2, 69115 Heidelberg 12 Prof. Dr. Knut Schäkel, 18.05.1966 PD Dr. Adelheid Cerwenka, 14.4.1968 Dept. Dermatology, MFH, Voßstr.2, 69115 Heidelberg Boveri Research Group Innate Immunity, DKFZ, INH 280, 69120 Heidelberg 13 Dr. Astrid Schmieder 05.02.1979 Prof. Dr. Viktor Umansky 23.12.1955 Dept. Dermatology, MFM; CCU Dermato-Oncology, DKFZ/MFM; UMM, 68135 Mannheim 1 Vascular Biology, Metastasis Fon: 0621-383-4461 Fax: 0621-383-3815 [email protected] Dept. Signal Transduction and Growth Control, DKFZ, INH 280, 69120 Heidelberg Dept. Dermatology, MFM, UMM, 68135 Mannheim, Germany RESEARCH AREA Signal Transduction, Tumor Progression Dermatology, Vascular Biology, Neurology, Brain Tumors Dermatology, DermatoOncology, Vascular Biology Dermatology, Vascular Biology DermatoOncology; Programmed Cell Death DermatoOncology; Programmed Cell Death DermatoOncology; Immunology Dermatology, Immunology Dermatology Immunology MFM = Medical Faculty Mannheim, Heidelberg University; MFH = Medical Faculty Heidelberg, Heidelberg University; UMM = University Medical Center Mannheim; DKFZ = German Cancer Research Center; CCU = Clinical Cooperation Unit Dermato-Oncology Mannheim, DKFZ/MFM 1 INTRO PRO- The group of applicants for the Research Training Group (RTG) comprises 13 professors/lecturers and 6 junior principal investigators. In designing the structure of the RTG, a number of factors were taken into consideration: 1. The DFG encourages the support of scientists on their way up the academic career ladder. Therefore, 6 projects recruited brilliant younger scientists as primary PIs who are in transition to become independent group leaders, allowing them to take responsibility for projects in the RTG early in their career development; 2. In addition to the 12 PhD positions we are applying for, the two Medical Faculties of Heidelberg University and the German Cancer Research Center (DKFZ) will finance one additional PhD position and 8 MD fellowships, amounting to a total of 21 doctoral students. To guarantee adequate supervision of these students, 19 PIs was considered appropriate, given that each doctoral student will be assigned two supervisors; 3. The study program we envision demands a high teaching quality and has to cover a range of topics for which a broad basis of experts in clinical and basic science will be necessary. The consortium of PIs we have assembled provides the requisite coverage of these topics; 4. Dermato-Oncology is a structural research focus of both Medical Faculties of Heidelberg University and the DKFZ. The high number of participating departments in this HeidelbergMannheim Skin Cancer Alliance allows, but also dictates that a higher number of lecturers than is recommended by the DFG for RTGs are included in this application. 1.4 Associated Researchers/Lecturers NAME, ACADEMIC TITLE 1 DEPARTMENT, AFFILIATION , POSTAL ADDRESS INTRO Prof. Dr. ClausDetlev Klemke Dept. Dermatology, MFM, UMM, 68135 Mannheim Prof. Dr. Wiebke Ludwig-Peitsch Dept. Dermatology, MFM, UMM, 68135 Mannheim Prof. Dr. Karsten Mahnke Dept. Dermatology, MFH, 69115 Heidelberg Prof. Dr. Hugo H. Marti Dept.Physiology,Neurovascular Research, MFH, 69120 Heidelberg Dr. Martin Sprick Hi-Stem gGmbH im DKFZ, 69120 Heidelberg CONTACT (FON, FAX, EMAIL) Fon: 0621-383-3918 Fax: 0621-383-3815 [email protected] Fon: 0621-383-1054 Fax: 0621-383-3815 [email protected] Fon: 06221-56-8170 Fax: 06221-56-1617 [email protected] Fon:06221-54-4138 Fax:06221-54-4561 [email protected] Fon: 06221-42-3913 Fax: 06221-42-3902 [email protected] RESEARCH AREA Dermatology, Skin Surgery, cutaneous lymphoma Dermatology, Rare Skin Cancers Immunology Vascular Physiology, Blood Brain Barrier Cancer stem cells, primary culture models of solid tumors 1.5 Summary in English and German 1.5.1 Summary in English Skin cancer constitutes a world-wide health issue of increasing importance as its incidence is continuously rising due to environmental factors and the aging population. Malignant melanoma is associated with a high mortality due to its tremendous metastatic potential. A similar number of deaths are caused annually by non-melanoma skin cancer, which is the most frequent cancer worldwide. As considerable therapeutic optimism has been recently raised by the development of designer drugs that target oncogenic signalling pathways and the tumor immune escape in skin cancer, the RTG focuses on a timely and evolving field of innovative research with a major socioeconomic impact on Western societies. The RTG will contribute considerably to a better understanding of skin cancer biology by addressing burning questions such as the molecular and cellular mechanisms (1) of skin cancer dissemination and metastasis, including skin cancer stem cells and the tumor vasculature and (2) of primary skin cancer resistance to apoptosis and immunity. All projects aim at identifying and validating novel therapeutic targets towards these hallmarks of skin cancer. The RTG research program builds on the intense collaboration of the applicants’ laboratories in the Heidelberg-Mannheim Skin Cancer Alliance whose collective expertise in skin cancer biology is unique in Germany. 2 The educational program is aimed at attracting young scientists to a hitherto under-developed field of research. The PhD/MD students will be trained in a broad portfolio of research skills and will also receive complementary teaching to increase their clinical knowledge. The RTG will closely collaborate with the existing Life Science Graduate Schools of Heidelberg University, and of the German Cancer Research Center (DKFZ) to recruit the best PhD students worldwide, and to provide teaching in basic molecular and cellular biology, as well as in general cancer biology and oncology. The added value of the RTG teaching program will lie in applying these general topics to the specifics of skin cancer biology and dermato-oncology. In addition, the RTG will support the PhD/MD students to conceive and realize scientific and teaching initiatives of their own, such as student project development platforms and student conferences. This will enable them later to develop their own research agenda in the field. In addition, the RTG will profit from the close scientific collaboration set up with the St. John’s Institute of Dermatology, King’s College, London, UK, one of the most renowned academic institutions in clinical and experimental dermatology worldwide. Beyond the St. John’s Institute, the RTG includes the participation of an inter-institutional, University of London and Cancer Research UK-based faculty of scientific project partners in the Metropolitan Area of London with outstanding expertise across the spectrum of relevant basic, translational and clinical science. The London project partners are committed to the goals of the RTG and will engage in pushing the scientific projects forward and help the PhD students develop their scientific carreers in an international environment. Altogether, the RTG will perform high quality research projects and train PhD and MD students to work closely together to fight skin cancer. The RTG will achieve its goals by combining basic science and clinical education with a focus on targetable hallmarks of skin cancer. Bösartige Hauttumoren stellen aufgrund von Umweltfaktoren und aufgrund der Altersentwicklung der Bevölkerung ein zunehmendes Gesundheitsproblem dar. Das maligne Melanom ist aufgrund seines ausgeprägten Metastasierungspotentials mit einer hohen Mortalität vergesellschaftet. Die epithelialen Hauttumoren sind die weltweit häufigsten bösartigen Tumoren mit einer vergleichbaren Mortalität. An den Therapieerfolgen der neuen zielgerichteten Medikamente bei bösartigen Hauttumoren wird deutlich, dass das GRK seinen Schwerpunkt auf ein hochaktuelles Thema legt. Das GRK wird zum besseren Verständnis der Biologie des Hautkrebses beitragen; Hauptthema sind die molekularen und zellulären Mechanismen (1) der Tumorzelldissemination und Metastasierung einschl. Hautkrebsstammzellen und Tumorgefäße sowie 2) der primären Resistenz gegenüber Apoptose und Tumorimmunabwehr. Jedes Projekt arbeitet zudem an der Identifizierung neuer therapeutischer Zielstrukturen bei diesen „Hallmarks of Skin Cancer“. Das RTG verstärkt die Zusammenarbeit zwischen den Arbeitsgruppen der Antragsteller, deren gemeinschaftliche Expertise zum Thema Hautkrebs in Deutschland ihres gleichen suchen dürfte. Das Qualifizierungsprogramm des GRK soll junge Forscher für ein bisher unterentwickeltes Gebiet begeistern. Die Graduierten werden eine breite methodische Ausbildung in der Grundlagenforschung und einen umfassenden Überblick über die klinische Dermato-Onkologie erhalten. Das GRK wird mit den Graduiertenschulen der Universität Heidelberg, HBIGS, und des DKFZ, HIGS, eng zusammenarbeiten. HBIGS wird für die Lehre in Molekular- und Zellbiologie, HIGS in Tumorbiologie und Allgemeiner Onkologie verantwortlich sein. Der Mehrwert des GRK wird in der Anwendung dieser Grundlagen auf die Biologie des Hautkrebses und die DermatoOnkologie liegen. Das GRK wird die Studenten dabei unterstützen, in eigener Verantwortung Forschungs- und Lehrinitiativen wie studentische Projektentwicklungsplattformen und Fachtagungen zu realisieren. Dies soll sie befähigen, später ein eigenes Forschungsprogramm in der Dermato-Onkologie zu entwickeln. Darüber hinaus wird das Forschungsprogramm und die Karriereentwicklung der Studierenden durch die Zusammenarbeit mit dem St. John’s Institute of Dermatology in London, einer der bekanntesten Forschungseinrichtungen der Dermatologie, an Internationalität gewinnen. Zusätzlich kann sich das GRK auf eine interinstitutionelle Gruppe von herausragenden Forscherpersönlichkeiten der verschiedenen Colleges der University of London und von Cancer Research UK stützen, die das Forschungssprogramm bereichern und ihre Expertise in den Dienst des GRK stellen werden. 3 INTRO 1.5.2 Summary in German Zusammenfassend wird das GRK hochqualitative Forschungsprojekte durchführen und die DoktorandInnen zur interdisziplinären Zusammenarbeit im Kampf gegen den Hautkrebs anleiten. Das GRK wird gerade durch die Verknüpfung von Grundlagenforschung und kliniknaher Ausbildung seine Ziele bei der Entwicklung neuer Therapien beim Hautkrebs erreichen können. 1.6 Funding period 01.04.2015-30.09.2019 1.7 Number of PhD and MD positions For a clinical subspecialty such as Dermato-Oncology, it is highly important to attract enthusiastic young scientists and to stimulate them to develop a long-term scientific career and ultimately an independent basic research agenda in the field. In addition, future clinical researchers in DermatoOncology will tremendously profit from additional expertise in the basic sciences. Therefore, the integration of both PhD and MD students into the RTG will be strongly supported. The interaction between PhD students and MD students will foster the mutual understanding between basic and translational research in Dermato-Oncology. MD students will bring a clinical twist into the RTG and help the PhD students with whom they work together to get a complete picture of the clinical background and specific questions of their projects. At the same time the PhD students will ensure transfer of knowledge and efficient training of MD students in a broad spectrum of basic research approaches and methodologies. As MD students who are willing to devote 1 year to experimental work in the laboratory are the exception rather than the rule, the RTG will restrict the number of MD fellowships to recruit only outstanding MD students with the goal of an academic career. Within the RTG, we apply for 12 PhD positions (0.65 TVL E13) within 13 projects over a period of 4.5 years. An additional PhD position will be financed by the DKFZ. Furthermore, a total of 8 MD fellowships will be funded by the medical faculties of Heidelberg University (5 MFM, 2 MFH) and the DKFZ (1 DKFZ) with a stipend of € 670,-- per month. Associated PhD and MD students. As all the applicants’ laboratories have research programs with a focus on skin cancer, a conservative estimate is that one associated PhD/MD student from each lab will participate in the RTG, making up a total of 13 associated student members. The associated researchers / lecturers will recruit additional associated PhD/MD students into the RTG. 2 Profile of the Research Training Group INTRO Heidelberg University and the German Cancer Research Center (DKFZ) together have developed a joint research and structural focus in the field of Dermato-Oncology. The departments of Dermatology in Heidelberg and Mannheim, the Clinical Cooperation Unit Dermato-Oncology of the DKFZ in Mannheim as well as the Department of Signal Transduction and Growth Control of the DKFZ make up the core of this Mannheim-Heidelberg Skin Cancer Alliance. Additional groups from both faculties and the DKFZ that have a major research interest in skin cancer biology further enhance and complement the consortium, including the Department for Vascular Biology and Tumor Angiogenesis, and the Division of Signaling and Functional Genomics. As skin cancer constitutes a world-wide health issue of increasing importance due to environmental factors and the aging population, the Dermato-Oncological Community in Heidelberg/Mannheim has committed itself to improve research, intensify scientific collaboration and attract young researchers to this evolving field. In order to prepare the RTG application, a Steering Committee convened consisting of Prof. Dr. H. Augustin, Prof. Dr. P. Angel, Prof. Dr. M. Boutros, Prof. Dr. A. Enk, Prof. Dr. S. Goerdt, Prof. Dr. M. Leverkus, Prof. Dr. S. Schneider, and Prof. Dr. J. Sleeman. After announcement of the planned RTG, 21 project proposals were submitted. Of these, 13 were selected for the RTG application. In 6 projects, brilliant younger scientists who are in the process of becoming independent group leaders were recruited as PIs to take project responsibility in the RTG early in their careers. These scientists will be encouraged to publish the results of their projects as senior authors, while being mentored by experienced senior scientists to assure successful project guidance. Of the 13 projects, 6 projects are University-based (4 Mannheim, 1 Heidelberg, 1 Mannheim / Heidelberg), 6 are dual University/DKFZ projects, and 1 project is DKFZ-based, resulting altogether in a wellbalanced collaboration between University and DKFZ groups. 4 3 Research Program 3.1 Overview and Aims Skin cancer is a rising socio-medical and economic threat to patients and the public. Due to environmental, behavioral, and demographic factors, the incidence of malignant melanoma (MM) and of non-melanoma skin cancer, especially cutaneous squamous cell carcinoma (SCC), is continuously rising in Caucasian populations. This is due to the increasing life expectancy and the intensified sun exposure in Western countries in the second half of the 20th century. SCC develops in a well-documented multi-step tumorigenesis process induced by UV damage from precursor lesions to fully malignant tumors. Precursor lesions (p53-mutated “patches”, actinic keratosis or SCC in situ) may still succumb to cell death and to the body’s own tumor immune responses. Much less is known about the carcinogenesis of MM, although molecular alterations in oncogenes and tumor suppressor genes and associated signaling pathways such as e.g. B-Raf may play a major role. While thin primary MM or SCC are usually treated in a curative manner by skin surgery, thick primaries have a high potential for loco-regional, lymph node and/or distant metastases that even if treated by standard therapeutic approaches will result in an unacceptably high mortality. In addition, treatment of skin cancer is counter-acted by primary resistance to cell death and tumor immunity. Therefore, there is an increasing need to better understand the mechanisms leading to skin cancer metastasis and to identify targets to overcome primary resistance. 5 INTRO The topic of the RTG “Hallmarks of Skin Cancer” is innovative, focused and timely. The research program of the RTG concentrates on elucidating the hallmarks of skin cancer, especially the intercommunicative rather than the cell autonomous qualities of tumors, such as cancer cell dissemination and metastasis as well as primary resistance towards cell death and immunity. These important topics will be studied with respect to the specifics of skin cancer, i.e. malignant melanoma and squamous cell carcinoma. Thus, the title of the RTG “Hallmarks of Skin Cancer – Cancer Cell Dissemination, Primary Resistance, Novel Targets” well reflects the focus of the research program of the RTG. To guarantee coherence within the RTG research program and to further enhance collaboration within in the RTG, the two major research areas, i.e. Cancer Cell Dissemination (A) and Primary Resistance (B), were subdivided into 4 project packages with 3-4 single projects relating to certain hallmarks of skin cancer, i.e Cancer Stem Cells (A1), Invasion and Metastasis (A2), Primary Resistance to Cell Death (A3), and Primary Resistance to Tumor Immunity (A4). As a cross-sectional aim, all projects have committed themselves to inherently direct their research towards the identification and validation of novel therapeutic targets. With respect to the teaching and qualification program, the areas covered by the projects allow for a sound interdisciplinary training of the graduate students. All participating laboratories have an impressive track record in skin cancer research and have proven their potential for innovative research by high quality publications. The combination of clinical disciplines with basic research laboratories guarantees sound experimental approaches with a clear translational focus. The study program of the RTG will profit from the collaboration with the Life Science Graduate Schools of Heidelberg University (Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology (HBIGS)) and of the German Cancer Research Center (Helmholtz International Graduate School for Cancer Research (HIGS)). HBIGS will be responsible for teaching and training the PhD/MD students in general molecular and cellular biology and in “soft skills”, whereas HIGS will be responsible for the teaching program in cancer biology and general oncology. The RTG will provide added value for the PhD/MD students by applying the general principles and approaches of cancer biology and oncology (the hallmarks of cancer) to the specific problems and questions of skin cancer biology and dermato-oncology (the hallmarks of skin cancer). Beyond the structured study program that accompanies the thesis work of every doctoral student, the collaboration with the St. John’s Institute of Dermatology in London, UK, and with an interinstitutional, University of London and Cancer Research UK-based faculty guarantees the internationality of the RTG and allows students as well as supervisors to have their scientific project approaches cross-checked by high ranked foreign researchers. Altogether, the innovative research program, the support by the Heidelberg Mannheim Skin Cancer Alliance, the high qualification of the participating researchers and the high-ranking international scientific collaboration will help the RTG live up to the high goals of a DFG-funded RTG program. Hanahan and Weinberg have claimed that six hallmarks of cancer are important for all malignant tumors including proliferation (mutations of oncogenes), evading growth inhibition (mutations of tumor suppressor genes), replicative immortality, invasion and metastasis, angiogenesis, and resistance to cell death. The former three hallmarks of cancer are executive, i.e. they occur within the cell and bestow the cancer cell with cellular autonomy. The latter three hallmarks are intercommunicative, i.e. tumorigenesis is supported by interactions of the cancer cell with noncancer cells. Recently, Hanahan and Weinberg have delineated two more executive hallmarks of cancer (genome instability and mutation, cellular energetics) and two more intercommunicative hallmarks of cancer (tumor immune escape, tumor-promoting inflammation). They especially emphasize (1) that it is necessary to analyze the significance of any hallmark of cancer in the context of a specific tumor entity, and (2) that the importance of the tumor microenvironment is becoming increasingly apparent. Therefore, the RTG (1) proposes to analyze hallmarks of skin cancer, and (2) focuses on intercommunicative hallmarks and the tumor microenvironment. Thus, the research program of the RTG aims to elucidate the cellular/molecular pathways that lead to skin cancer cell dissemination (Research Area A), and to analyze the mechanisms of primary skin cancer resistance to cell death and tumor immunity (Research Area B). In a cross-sectional approach, all projects aim to identify novel targets against the hallmarks of skin cancer they study. INTRO 3.1.1 Research Area A – Cancer Cell Dissemination Project Package A1: Cancer Stem Cells (Projects 1-4) Project Package A2: Invasion and Metastasis (Projects 5-7) The processes leading to skin cancer dissemination and metastasis are complex and far from being completely understood. The RTG will therefore aim to investigate in depth the molecular and cellular mechanisms of skin cancer dissemination and metastasis (Projects 1-7). As in other cancers, metastasis in MM and SCC follows an ordered sequence of events. However, the MMand SCC-specific molecular and cellular determinants of these events may vary as predicted by the “seed and soil” hypothesis, and only a minor population within all tumor cells seems to be involved in metastasis. The concept of cancer stem cells (CSC) and their potential plasticity is therefore of great importance. Counter-intuitively, CSC may already acquire invasive potential and metastasize early during tumorigenesis, causing the phenomenon of tumor dormancy. CSC may orchestrate different waves of cancer cell dissemination from the primary as well as from different distant metastatic foci and may even re-colonize the primary (Projects 1-3). As a precondition for dissemination, MM and SCC cells have to acquire enhanced cell motility and invasive capacity that is regulated by signaling pathways (Project 3, 4) and mediated by adhesion molecules (Project 5). These steps are followed by intravasation and transit via the blood vascular or lymphatic systems. The circulating tumor cells need to be arrested intravascularly at the site of metastatic colonization, e.g. by binding to coagulation factors deposited on the luminal side of the vessel wall (Project 6) or by binding to organ-specific endothelial adhesion molecules (Project 7). Subsequently, tumor cells must extravasate and finally grow at the distant site. During this phase, the tumor cells need strong survival mechanisms, and must resist immune surveillance (Research Area B). 3.1.2 Research Area B – Primary Resistance to Cell Death and Immunity Project Package B1: Primary Resistance to Cell Death (Projects 8-10) Project Package B2: Primary Resistance to Tumor Immunity (Projects 11-13) Programmed cell death by apoptosis has been recognized as an important barrier towards the development of various cancers. Conversely, compelling evidence has accumulated that resistance to cell death occurs in malignant tumors as they develop into high grade malignancies, and accompanies resistance to treatment in general. Resistance to cell death has been shown to develop in both MM and SCC. However, the first therapeutic trials with drugs blocking Bcl-2 familiy members have failed, indicating the need for a better understanding of cell death resistance in skin cancer. Programmed cell death is a sophisticated multi-facetted process. The RTG research program includes three projects dealing with new aspects of programmed cell death. Project 8 investigates protection of MM cells against anoikis, a special form of apoptosis caused by inadequate tumor cell-matrix interactions that may be targetable for therapy by angiopoietin-2 antagonists/antibodies. Project 9 analyzes a novel form of programmed cell death in SCC called programmed necrosis or necroptosis, and its regulation by the ripoptosome, a newly identified 6 subcellular cell death-associated platform. Finally, project 10 examines the potential of a novel class of small molecules called IAP antagonists to overcome resistance to cell death in MM. An important form of eradication of incipient neoplasias, late-stage tumors or micrometastases is immune-mediated cell death. Immunosurveillance is active in skin cancer in both SCC and MM as immunocompromised patients; for example, show a tremendous tendency to develop high grade SCC. However, immunosurveillance seems to be effective only in incipient SCC; with progression, SCC and MM develop mechanisms of tumor immune escape or even hi-jack the immune system for their needs. Project 11 will analyze the role of the chemokine receptor CCR6 in immunosurveillance of MM. Project 12 will study how pro-inflammatory cytotoxic dendritic cells may be used to re-direct the host immune system towards efficient immunological tumor cell killing. Finally, there is a growing body of evidence that tumor-associated myeloid cell populations are educated by the tumor itself to support tumor growth and metastasis (Project 13). 3.1.4 Collaboration, Methods, Model Systems Within the RTG, the projects will intensively collaborate. Examples are the transfer of novel targets to unique model systems, such as mouse models (Project 5,13), human skin cancer stem cell models (Project 1,2), and imaging (Project 6). The details of the collaborative interactions between the projects are outlined in the project descriptions. The experimental methods used include molecular and cell culture techniques, state-of-the-art gain- and loss-of function approaches including CRISPR/Cas9 genome editing, large scale RNAi screening, novel model systems in human cells, and additional cutting edge technologies in skin cancer research. Furthermore, animal models including the ret transgenic MM mouse (e.g. Projects 6,7,13), the well-established DMBA/TPA SCC model (Project 5), and the transgenic K5-SOS-F transgenic mouse are available. As a result, a broad spectrum of molecular, genetic, biochemical, cell biological, histological and morphometrical techniques will be ready to use within the RTG. The entire RTG including the associates will benefit from this expertise, both in terms of education and in terms of competitive research. 7 INTRO 3.1.3 Novel Targets – A Cross-sectional Approach Targeting known molecular and cellular events that regulate tumor progression and metastasis in MM and SCC has led to remarkable therapeutic advances in the recent years. Unfortunately, these treatment modalities provide only short term clinical remissions due to secondary resistance development. As a result, there is a need to identify additional targets for tumor progression, metastasis, and primary tumor cell resistance to hit skin cancer with combination therapies before secondary tumor cell resistance has developed. Therefore, the identification of novel targets in skin cancer will be a cross-sectional focus of the RTG. Several projects will investigate tumor-specific targets in MM or SCC, including podoplanin or Wnt in invasion by SCC (Project 4,5), molecules mediating intravascular MM cell arrest (Projects 6,7), the inhibition of cell death pathways in MM and SCC (Projects 8-10) and the generation of cytotoxic dendritic cells (Project 12). In order to validate novel targets, human model systems of the metastatic process are needed (Project 6). The combination of projects covering individual targets with other projects that develop innovative methodology will allow novel targets to be directly tested synergistically within the framework of the RTG. 3.2 Project Descriptions Research Area A – Cancer Cell Dissemination Project Package A1: Cancer Stem Cells (Projects 1-4) Project 1: The role of Id proteins in determining the tumor initiating and metastatic properties of melanoma cells Principal Investigator: Prof. Dr. Jonathan Sleeman, CBTM, Medical Faculty Mannheim, Heidelberg University London Project Partner: Dr. Caroline Hill, Cancer Research UK, London Research Institute Short Summary Cancer stem cells are thought to underpin the growth, metastasis and therapy resistance of tumors such as melanoma, through their tumor initiating properties. Building on unpublished observations, this project aims to substantiate the hypothesis that Id gene expression induced by 3D extracellular matrix (ECM) microenvironments plays a functional role in determining stemness and metastatic properties of melanoma cells. The role of integrins, TGF-β and BMPs in determining these properties will be determined. These data should identify new therapeutic targets. PACKAGE A1 3 State of the Art 3.1 State of knowledge in the field The concept that the bulk of cells that make up a tumor, including melanomas, are derived from cancer stem cell (CSC) subpopulations is now widely accepted. CSCs are distinguished from other tumor cells by their ability to successfully seed new tumors when implanted in low numbers into experimental animals, and to recapitulate the morphology of the initial tumor. In contrast, the nonCSC population cannot initiate tumor growth in vivo even when implanted in high numbers. The potential significance of CSCs for cancer therapy is enormous. Current therapies appear to preferentially destroy the non-CSC population but do not efficiently kill CSCs, with the result that the tumor eventually regrows. Furthermore, as the CSC subpopulation represents by definition the only tumor cells that are able to initiate the growth of new tumors, then CSCs must play a central role in metastasis formation. Understanding the parameters that determine tumor-initiating properties should therefore identify targets for novel and efficient cancer therapies. Tumor initiation in vivo is used to define CSCs. Numerous papers have shown that the take rate of tumors in vivo can be manipulated, for example by coinjecting tumor cells with matrigel. Indeed, single unsorted human melanoma cells in matrigel are capable of forming tumors in NOD/SCID Il2rg-/- mice (Quintana et al., 2008, Nature). These observations point to a critical role for the microenvironment, in particular the extracellular matrix, in determining tumor-initiating properties. 3.2 Preliminary work by the participants We have a long track record in metastasis research. Recently we have begun exploring the role of CSCs in metastasis, partly in collaboration with Prof. Umansky (Project 13), with whom we have published work using the Ret murine melanoma model. In unpublished work we have investigated the role of ECM components in determining tumor-initiating properties in vivo. As few as 5 cells from the B16 or Ret murine melanoma cell lines were sufficient to initiate tumor growth when coinjected into syngeneic mice with matrigel. In contrast, tens of thousands of cells were required to initiate tumor growth in the absence of matrigel. In further experiments we found that co-injection of tumor cells with laminin or with collagen type I was also sufficient to elicit tumor growth from 5 cells. These data indicate that highly immunocompromised mice are not required for tumor initiation from just a few cells, and that several ECM components are able to initiate tumor growth from small numbers of melanoma cells, all of which are ligands for β1-containing integrins. Microarray analysis revealed that Id1, Id3 and Smad6, archetypal TGF-β/BMP response genes, were uniquely upregulated (up to 40-fold) in response to 3D but not 2D ECM (matrigel, collagen or laminin), a finding confirmed using qPCR. Id genes are known to play a pivotal role in regulating tumor growth and determining stemness properties, while Smad6 counter-regulates TGF-β/BMP 8 signalling. We also found that tumor cells that do not respond to 3D ECM by upregulating Id1 and Id3 do not show efficient tumor initiation when co-injected with matrigel in vivo. Collaborative work with Prof. Utikal (Project 2) has shown that human melanoma cells can also respond to 3D matrix by upregulating Id1, Id3 and Smad6. 4.2 Experimental program Aim 1: We will establish loss of function (shRNA, cannabidiol chemical inhibition) and gain of function (tet-inducible expression) for Id1 and Id3 (either alone or in combination) in B16 and Ret melanoma cells. We will then test the cells for their tumor initiating and metastatic ability in vivo in the presence (loss of function) or absence (gain of function) of matrigel. These data will demonstrate whether Id1 and Id3 induction in 3D ECM plays a role in specifying the tumor-initiating and metastatic properties of melanoma cells. Aim 2: We will use loss of function (shRNA) and gain of function (constitutively active mutant) approaches to determine whether β1-containing integrins are involved in the induction of Id1, Id3 and Smad6 in response to 3D ECM microenvironments. If so, we will determine using B16 and Ret cells whether loss of β1 ablates efficient tumor initiation in vivo in the context of 3D ECM, and what effect it has on metastasis. Conversely, we will establish whether constitutively activated β1 integrin supports efficient tumor initiation in vivo even in the absence of a 3D ECM matrix, as well as whether it promotes metastasis formation. Aim 3: We will determine which members of the TGF-β and BMP families and their receptors are expressed in B16 and Ret cells growing in 3D ECM environments. For those that are expressed, we will use shRNA to knockdown their expression, and/or use specific inhibitors of TGF-β and BMP signalling to block their activity, then determine whether this affects the ability of 3D ECM to induce Id1, Id3 and Smad6 expression, and to promote efficient tumor initiation in vivo. Effects on metastasis formation will also be determined. We will also investigate whether 3D ECM environments promote Id1, Id3 and Smad6 expression by sequestering TGF-β and BMP family members produced by the melanoma cells at locally high concentrations in the ECM around the cells. For all aims, the results of the experiments will be corroborated in human melanoma cells to demonstrate relevance to human disease. 4.3 Collaborations with other projects in the RTG: Collaborations with Prof. Utikal (Project 2) will include (i) isolation of CSC marker-enriched subpopulations from human melanoma samples and analysis of their Id protein expression (ii) analysis of Id expression in primary human melanomas and their metastases. Collaborations with Projects 5 and 7 comprise provision of antibodies, genetically modified mice, and expertise in techniques such as the analysis of cell interactions with hyaluronan and lymphangiogenesis. 5 References 1. Müller T, Stein U, Poletti A, Garzia L, Rothley M, Plaumann D, Thiele W, Bauer M, Galasso A, Schlag P, Pankratz M, Zollo M, Sleeman JP. 2010. ASAP1 promotes tumor cell motility and invasiveness, stimulates metastasis formation in vivo, and correlates with poor survival in colorectal cancer patients. Oncogene 29:2393–2403 2. Neeb A, Wallbaum S, Novac N, Scholl I, Dukovic-Schulze S, Schreiber C, Schlag P, Moll J, Stein U, Sleeman JP. 2012. The immediate early gene Ier2 promotes tumor cell motility and metastasis, and predicts poor survival of colorectal carcinoma patients. Oncogene 31:3796-806 3. Kuch V, Schreiber C, Thiele W, Umansky V, Sleeman JP. 2013. Tumor initiating properties of breast cancer and melanoma cells in vivo are not invariably reflected by spheroid formation in vitro, but can be increased by long-term culturing as adherent monolayers. Int J Cancer 132:E94-105. 9 PACKAGE A1 4. Project Plan 4.1 Specific Aims: (1) To test the hypothesis that Id1 and Id3 expression induced in 3D ECM microenvironments plays a functional role in determining the tumor initiating and metastatic properties of melanoma cells; (2) To determine whether β1-containing integrins mediate 3D ECMmediated upregulation of Id1, Id3 and Smad6, and are required for efficient tumor initiation and metastasis formation in vivo; (3) To investigate whether TGF-β and/or BMP signalling mediates increased Id1, Id3 and Smad6 in 3D ECM microenvironments. Project 2: Characterization of human melanoma cells on the basis of markers of pluripotent stem cells Principal Investigator: Prof. Dr. Jochen Utikal, Skin Cancer Unit, German Cancer Research Center and Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Ruprecht-Karl University of Heidelberg London Project Partner: Prof. Fiona Watt, Centre for Stem Cells and Regenerative Medicine, King’s College, London Short Summary Embryonic stem cells (ES cells) are similar to melanoma cells in many aspects. ES cells are immortal and proliferate rapidly. They also form tumors (teratomas) when transplanted into immune-deficient mice. Similar to ES cells melanoma cells show also a plasticity. By the ectopic overexpression of different sets of transcription factors or microRNAs somatic cells can be converted into ES-like cells. Our studies indicate that melanoma comprise different subpopulations which express marker of pluripotent stem cells (e.g. Nanog, Sox2). The projects objective will be to identify such subpopulations within murine and human primary melanoma cells as well as in melanoma cell lines. Respective subpopulations will be analyzed at the genomic, epigenomic, and proteomic level. Functional abilities for maintaining the tumor cell growth will be tested. Moreover, the accessibility of different subpopulations towards cellular reprogramming, the reprogramming kinetics and factor requirements will be investigated. This project should help to understand the maintenance and formation of melanomas. The examined markers might serve as therapeutic targets in future. 3 State of the Art 3.1 State of knowledge in the field Embryonic stem cells (ES cells) are similar to tumor cells in many aspects. ES cells are immortal and proliferate rapidly. They also form tumors (teratomas) when transplanted into immune-deficient mice. PACKAGE A1 3.2 Preliminary work by the participants We have shown that different cell types including mouse and human melanocytes or melanoma cells can be reprogrammed into pluripotent stem cells by the ectopic expression of transcription factors such as Oct4, Klf4, Sox2 and c-Myc. These pluripotent stem cells have all the features of ES cells including immortal growth, the expression of pluripotency markers (e.g. Sox2 and Nanog) and the potential of forming teratomas (Stadtfeld et al., 2008; Eminli et al., 2008; Utikal et al., 2009a). The conversion efficiencies of melanocytes into pluripotent stem cells can be increased dramatically by downregulating p53 or p16/p19 further underscoring similarities of this mechanism with tumorigenesis (Utikal et al., 2009b). Our preliminary studies show that subpopulations of human melanoma cells reveal an endogenous expression of pluripotency markers such as Nanog or Sox2. However, these cell populations are not yet well characterized and their functional abilities for maintaining the tumor cell growth are not yet known. 4 Project Plan 4.1 Specific Aims Main hypothesis: Markers of pluripotent stem cells play a main role in development and maintenance of human malignant melanoma Aim 1: Identification of cell populations which express markers of pluripotent stem cells (e.g. Sox2, Nanog) in murine and human primary melanoma cells and melanoma cell lines. Aim 2: In-depth analysis and comparison of expression profile and epigenetic status as well as functional analysis of different subpopulations. 10 4.3 Collaborations with other Projects in the RTG For the analysis of melanoma subpopulations at the genomic, epigenetic, and proteomic level, we will closely collaborate with projects 1 and 4. Functional abilities for initiating and maintaining tumor cell growth will be tested in cooperation with project 10. The ret transgenic mouse model of malignant melanoma will be provided by project 13. 5 References 1. Eminli S*, Utikal J*, Arnold K, Jaenisch R, Hochedlinger K. 2008. Reprogramming of neural progenitor cells into induced pluripotent stem cells in the absence of exogenous Sox2 expression. Stem Cells 26:2467-74. * authors contributed equally 2. Stadtfeld M, Nagaya M, Utikal J, Weir G, Hochedlinger K. 2008. Induced pluripotent stem cells generated without viral integration. Science 322:945-9. 3. Utikal J, Maherali N, Kulalert W, Hochedlinger K. 2009. Sox2 is dispensable for the reprogramming of melanocytes and melanoma cells into induced pluripotent stem cells. J Cell Science 122:3502-10. 4. Utikal J, Polo JM, Stadtfeld M, Maherali N, Kulalert W, Walsh RM, Khalil A, Rheinwald JG, Hochedlinger K. 2009. Immortalization eliminates a roadblock during cellular reprogramming into iPS cells. Nature 460:1145-8. 11 PACKAGE A1 4.2 Experimental program In order to identify subpopulations of melanoma cells expressing common markers of pluripotent stem cells such as Sox2 or Nanog lentiviral reporter constructs will be generated. Reporter constructs will be designed such that the promoter of a particular pluripotency marker will control the co-expression of a fluorescing (e.g. GFP) and an antibiotic selection marker. Lentiviral particles carrying the reporter constructs will be produced and purified. Primary melanoma cells directly isolated from patients, from our established transgenic RET melanoma mouse model as well as cells from melanoma cell lines (e.g. C32, HT144) will be infected with lentiviral vectors that carry the reporter constructs. Cells expressing the respective genes and accordingly also expressing the fluorescent marker will be visualized by means of fluorescent microscopy. Quantification and separation of fluorescently labelled Sox2- and Nanogexpressing melanoma cells will be done by fluorescence activated cell sorting. This will also enable to monitor if cells from a certain subpopulation have a stable phenotype or might perhaps convert to cells from a different subpopulation. The sorted populations will be compared by analysing the expression of additional stem cell markers, global gene expression and DNA methylation in detail. Immunofluorescent labeling will be performed to examine the expression of markers (e.g. Sox2, Nanog, SSEA-3/4, Lin28) and melanoma-specific markers (e.g. S100, MART-1, HMB-45, MITF), respectively. For the evaluation of differential gene expression DNA microarrays and RT-PCR will be performed. Moreover, the promoter methylation status of stem cell- and melanoma-associated genes as an indicator for transcriptional activity will be checked by bisulfite sequencing. To correlate the afore mentioned parameters with functional properties, the tumorigenic potential of subpopulations will be investigated. For this purpose, the cells will be injected subcutaneously into immune-deficient mice (in the case of human cells) or in the syngenic RET transgenic melanoma mouse model (mouse melanoma cells) and tumor growth will be quantified. By knocking down the expression of melanoma- and pluripotency-associated genes with shRNA, the role of these genes for tumor development and growth will be studied. The impact of different subpopulations on tumor development will be analyzed by specifically depleting those cells. This will be achieved by infecting melanoma cells with a lentiviral construct that contains a promoter (e.g. Nanog or Sox2 promoter) which controls the expression of the enzyme thymidine kinase. Application of ganciclovir will selectively deplete all cells which express thymidine kinase. Tumor growth in the absence and presence of different subpopulations will be compared and marker switching abilities of tumor cells will be investigated. Another functional aspect we will focus on is the amenability of melanoma subpopulations to the process of reprogramming by transcription factors such as ectopic Oct4, Klf4 and c-Myc. Reprogramming kinetics and factor requirements will be determined. Project 3: The role of Wnt signaling in tumor-initiating cells and tumor progression in cutaneous SCC Principal Investigator: Dr. Iris Augustin, Prof. Dr. Michael Boutros, Div. of Signaling and Functional Genomics DKFZ, and Dept. Cell and Molecular Biology, Medical Faculty Mannheim, Heidelberg University London Project Partner: Prof. Fiona Watt, Centre for Stem Cells and Regenerative Medicine, King’s College, London Short Summary The project is aimed at deciphering the role of autocrine and paracrine Wnt signaling in squamous cell carcinoma (SCC). To investigate the role of the Wnt ligands in papilloma formation and their progression to carcinomas, we will selectively ablate the Wnt secretion factor Evi in skin SCC cell lines and in keratinocytes of adult mice. The role of Evi in SCC will be addressed in cell-based assays as well as in transplantation experiments. To genetically manipulate Evi during SCC tumorigenesis, we will target different cellular compartments of the skin epidermis crossed with KRasLSL-G12D and TP53fl/fl mice as well as with Evifl/fl mice to dissect the contribution of Wnt signaling to tumor initiation and malignancy in IFE- and HF-derived cutaneous SCC. The project will thereby provide insights in cellular signal transduction controlling skin carcinogenesis. PACKAGE A1 3 State of the Art 3.1 State of knowledge in the field Keratinocyte-derived non-melanoma skin cancers (NMSC) comprising basal cell (BCC) and squamous cell carcinoma (SCC) are the most common malignancies. SCCs are locally invasive and acquire the ability to metastasize, making them suitable models to study tissue invasion and metastasis. SCC development follows a multi-step model of tumorigenesis including Ras and p53 mutations, and they may derive either from adult interfollicular (IFE) or follicular (HF) tumorinitiating cells (TIC). Wnt signaling is critically involved in tumor initiation and progression of various types of tumors including the fate of TICs. Yet, the downstream mechanisms of SCC tumorigenesis involving Wnt signaling have not been unraveled. Beta-catenin deletion suppresses TICs in SCC and induces tumor regression. Invasive SCC is also marked by the concurrent upregulation of ßcatenin-independent and repression of ß-catenin-dependent Wnt signaling. As such, it has been shown that Wnt5a gradients enhance directed motility of keratinocytes. Wnt5a deficiency suppresses tumor growth of human malignant HaCaT-II4 cells. Yet, the downstream mechanisms of SCC tumorigenesis involving Wnt signaling have not been unraveled. 3.2 Previous work by the participants Previous work of the applicant addressed the contribution of Wnt signaling in developmental and pathological conditions. The research focused on the Wnt secretion factor Evi/Wls. Evi is required for the secretion of all Wnt ligands and therefore essential for pan Wnt signaling cascades. Manipulating Evi function in a time- and cell type-specific manner represents a versatile tool to study autocrine and paracrine Wnt signaling in different biological contexts. Experiments on glioblastoma revealed an important role of Evi in glioma tumorigenesis (Augustin et al., 2012). Furthermore, we generated transgenic mouse lines, conditionally Evi-deficient or ectopically overexpressing Evi. These mice were analyzed regarding Evi function in skin homeostasis. Targeted deletion of Evi in keratinocytes revealed aberrant skin morphology together with enhanced immune cell recruitment, which closely resembled human psoriatic skin disorders (Augustin et al., 2013). 12 4 Project Plan 4.1 Specific Aims The proposed project is aimed at pursuing the following specific aims: 1. What is the role of Wnt secretion in malignant epidermal tumor initiation and progression? 2. What is the mechanistic contribution of Wnt secreting tumor cells in the maintenance of cutaneous tumor initiating cells (TIC)? 4.3 Collaborations with other Projects in the RTG Mouse model studies will be performed in collaboration with project 5. We will work with project 4 on the role of Wnt signaling in melanoma cells and with project 9 on SCC biology. We will study angiogenesis in resulting tumors in collaboration with project 8. 5 References 1. Augustin I, Gross J, Baumann D, Korn C, Kerr G, Grigoryan T, Mauch C, Birchmeier W, Boutros M. 2013. Psoriasiform dermatitis-related phenotype caused by loss of epidermal Wnt secretion. J Exp Med 26:1761-77 2. Voloshanenko O, Erdmann E, Dubash T, Augustin I, Metzig M, Hundsrucker C, Kerr G, Sandmann T, Anchang B, Demir K, Boehm C, Leible, Ball C, Glimm H, Spang R, Boutros M. 2013. Wnt secretion is required to maintain high levels of Wnt activity in colon cancer cells. Nat Commun 4:2610 3. Augustin I, Goidts V, Bongers A, Kerr G, Vollert G, Radlwimmer B, Hartmann C, Herold-Mende C, Reifenberger G, von Deimling A, Boutros M. 2012. The Wnt secretion protein Evi/Gpr177 promotes glioma tumourigenesis. EMBO Mol Med 4:38-51 13 PACKAGE A1 4.2 Experimental program Aim 1: Our previous results have shown that epidermally secreted Wnt ligands play important roles in skin homeostasis. In order to determine Evi function in malignant tumor growth, we will generate Evi-deficient murine SCC cell lines (BDVII, PDVA) by CRISPR/Cas9 technology and characterize their proliferation and mobility. Transwell experiments will be performed to analyze tumor cell migration and invasiveness. In order to determine the paracrine effect of Wnt secreting stromal cells on the migratory and invasive growth of SCC cells, Transwell co-culture assays with Evioverexpressing MEFs (Wnt-on) as well as Evi-deficient MEFs (Wnt-off) will be compared. Subcutaneous injection of these SCC cells in mice will be performed to analyze tumor growth, morphology and invasive behavior. Epidermal deficiency of Evi impairs the cross talk between keratinocytes and immune cells (Augustin et al., 2013). Therefore, transplantation of SCC cells in syngeneic and immune competent recipients will provide an additional tool to study tumor immune cell infiltration. Aim 2: To genetically manipulate Wnt signaling during SCC tumorigenesis in vivo, we will use hairfollicle-specific (K19creER) and inter-hair-follicle-specific (INVcreER) driver mice crossed with KRasLSL-G12D and TP53fl/fl mice as well as with Evifl/fl mice to dissect the contribution of Wnt signaling to tumor initiation and malignancy in IFE- and HF-derived cutaneous SCC. In order to decipher the role of Wnt secretion on TIC maintenance, TICs will be analyzed from multiple Evi loss-of-function transgenic animals. FACS-based isolation of TICs (CD34high/EpCAMhigh) will be performed and the cells will be expression-profiled. TIC transcript signatures provide insights in Wnt-dependent TIC signaling networks. This project will apply complex mouse genetics to study the role of Wnt secretion in TICs. The indicated experimental setup schedules the investigation of Evi function in malignant SCC cell lines prior the transgenic mouse studies. Therefore, the time for the breeding procedure is experimentally covered by the analyses of SCC cell line in transplantation and cell-based assays. Project 4: Identification of genes linked to aberrant Wnt secretion in melanoma Principal Investigator: Prof. Dr. Michael Boutros, Div. of Signaling and Functional Genomics DKFZ, and Dept. Cell and Molecular Biology, Medical Faculty Mannheim, Heidelberg University London Project Partner: Prof. Frank Nestle, St John's Institute of Dermatology, King’s College, London, and Prof. Buzz Baum, Laboratory for Molecular Cell Biology, Medical Research Council, University College London Short Summary Wnt signaling has been implicated in multiple stages of melanoma tumorigenesis. Recently, it has been shown that downregulation of Evi/Wls, the cargo-receptor for all Wnt ligands, has a profound effect melanoma cell proliferation and metastasis. Concordantly, melanoma biopsies showed a decrease in Evi levels and low levels of nuclear β-catenin. Here, we propose to dissect the cellular processes that are influenced by aberrant Evi levels in melanoma cells. We will perform genomescale RNAi screens to identify genetic interactors of Evi which act as synthetic lethals with loss-of or gain-of function Evi. We will further characterize these genes by biochemical and cell-biological methods. In addition, we will analyse the role of Wnt secretion of the microenvironment of melanoma cells ex vivo and in transgenic mouse models for melanoma in vivo. PACKAGE A1 3 State of the Art 3.1 State of knowledge in the field Wnt proteins are secreted morphogens that regulate key processes during development and homeostasis. Aberrant regulation by means of overexpression of ligands or signal transducers has been linked to many cancers. The secretion of Wnt ligands depends on the Wnt-specific cargoreceptor Evi/Wls which binds to Wnt proteins in the Golgi and is required for its transport to the plasma membrane and exocytosis (Bartscherer et al., Cell 2006). Evi is a cargo-receptor for both canonical and non-canonical Wnt ligand secretion and has been recently shown to be upregulated in glioma and required for glioma cell proliferation, migration and tumor initiation (Augustin et al., EMBO Mol Med 2012). Wnt signaling is implicated in the progression and metastasis of melanoma. Normal skin and benign nevi express several Wnt proteins (Wnt2, Wnt5a, Wnt7b, Wnt10b) which are downregulated in melanoma. In particular, decreased levels of canonical Wnt signaling has been correlated with a poor outcome. Concordantly, it was recently shown that loss-of Evi expression in melanoma cells leads to an increase in cell proliferation and metastasis (Yang et al., EMBO Mol Med 2012), however, the exact molecular mechanism remains poorly understood. 3.2 Previous work by the participants The Wnt cargo-receptor Evi was first discovered in our laboratory in a genome-wide RNAi screen in Drosophila (Bartscherer et al., Cell 2006) and has subsequently shown to have a conserved function in Wnt signaling throughout the animal kingdom. Evi is a specific cargo-receptor to Wnt proteins, as other signaling routes appear not to be affected. Deletion of Evi during mouse development leads to early embryonic phenotypes similar to Wnt3a. We have recently shown that Evi is overexpressed in astrocytic glioma and promotes glioma tumorigenesis (Augustin et al., 2012). Interestingly, while Evi acts as an oncogene in glioma, it has a tumorsuppressor function in melanoma, indicating context-specific roles, e.g. in the balance between canonical and noncanonical Wnt signaling. To study tissue-specific role of Wnt secretion in vivo, we have generated conditional Evi loss-of-function (Evi-LOF) and Evi overexpression transgenic (Evi-GOF) mice. EviK14-Cre knockout mice display, in addition to impaired hair follicle morphogenesis, an inflamed skin and prominent neutrophil infiltration followed by T cell recruitment, leading to chronic inflammation. 14 4 Project Plan 4.1 Specific Aims This project has two overall aims: (1) we intend to dissect the cell-autonomous regulatory circuits that contribute to Evi mediated proliferation and metastasis in melanoma. To this end, we will perform genetic interaction screens using melanoma cell lines that have either low or high levels of Evi expression, searching for genes that act synergistically or antagonistically with Evi expression. (2) We will analyze the identified candidate genes by biochemical and cell-biological methods. We are particular interested in genes that dysregulate Evi expression or act downstream to influence cell proliferation and metastasis. Taken together, these studies should provide a better understanding on the role of Wnt ligands and their downstream signaling during melanoma tumorigenesis. 4.3 Collaborations with other Projects in the RTG We will work with Project 4 on the role of Wnt signaling in melanoma cells. We further collaborate with project 9 (Leverkus) and 10 (Geserick/Leverkus) on high-throughput RNAi analysis and CRISPR/Cas9 technology. 5 References 1. Gross JC, Chaudhary V, Bartscherer K, Boutros M. 2012. Active Wnt proteins are secreted on exosomes. Nat Cell Biol 14:1036-45 2. Augustin I, Goidts V, Bongers A, Kerr G, Vollert G, Radlwimmer B, Hartmann C, Herold-Mende C, Reifenberger G, von Deimling A, Boutros M. 2012. The Wnt secretion protein Evi/Gpr177 promotes glioma tumourigenesis. EMBO Mol Med 4:38-51 3. Horn T, Sandmann T, Fischer B, Axelsson E, Huber W, Boutros M. 2011. Mapping of signalling networks through synthetic genetic interaction analysis by RNAi. Nature Methods 8:341-6 4. Buechling T, Chaudhary V, Spirohn K, Weiss M, Boutros M. 2011. p24 proteins are required for secretion of Wnt ligands. EMBO Rep 1:1265-72. 5. Bartscherer K, Pelte N, Ingelfinger D, Boutros M. 2006. Secretion of Wnt ligands requires Evi, a conserved transmembrane protein. Cell 5:523-33. 15 PACKAGE A1 4.2 Experimental program Aim 1: In order to identify synthetic genetic interactions (synthetic lethals) of Evi melanoma, we will conduct genome-scale RNAi screens using A375 and A2058 melanoma cell lines. Using CRISPR/Cas9 technologies, we will create isogenic cell lines that have high or low levels of Evi expression. Using high-throughput microscopy, we will score a range of phenotypes, including cell proliferation and viability, but other changes in overall cell morphology. This will be done in collaboration with the lab of Buzz Baum who is an expert in systems analysis of signaling and cell shape. Computational analysis will be performed as described in Horn (2011). Pathway specific analyses of identified candidate gene will indicate signaling circuits and might open the possibility to test selective inhibitors. Aim 2: Identified candidates will be validated and further studied to understand their biological role in cells and using mouse models in vivo. We expect that candidate genes can act as epigenetic repressors of Evi expression in melanoma or restore canonical Wnt signaling downstream of Wnt secretion. For epigenetic regulators, we will perform ChIP experiment to test whether they act on enhancers in the Evi locus. We will assess whether regulators of Evi expression are similarly dysregulated in melanoma. Candidate genes acting downstream of Wnt secretion will be analyzed for their activity in canonical and non-canonical signaling branches and will assess their effect on cell proliferation and metastasis ex vivo and in vivo. Research Area A – Cancer Cell Dissemination Project Package A2: Invasion and Metastasis (Projects 5-7) Project 5: Function of the mucin-like glycoprotein podoplanin in squamous cell carcinoma progression Principal Investigator: Prof. Dr. Peter Angel, Division Signal Transduction and Growth Control, German Cancer Research Center (DKFZ), Heidelberg London Project Partner: Dr. Joy Burchell, Research oncology KCL, Guy's Hospital, London Short Summary The mucin-like glycoprotein podoplanin (PDPN) represents a tumor-associated protein in a variety of tumors including squamous cell carcinoma of the skin in mouse and human and correlates with poor prognosis and metastatic risk. Overexpression of PDPN in pancreatic cancer promoted tumor cell invasion and affected migration of glioblastoma and keratinocyte cell lines via modulation of the cytoskeleton pointing to a fundamental role of PDPN in tumor cell migration and invasion. We will i) measure transcriptional control of PDPN by altered cell death pathways promoting SCC formation and ii) define the in vivo function of PDPN in SCC. Here, we will apply loss-of-function approaches in cultured human SCC cells with metastatic potential and measuring parameters of cell invasion and actin cytoskeleton. Importantly, we will make use of floxed PDPN mice recently generated in our lab to specifically delete this gene in keratinocytes via K14-Cre transgenic mice. To unequivocally define the function of PDPN in tumor development and progression, the well established chemically induced in vivo skin carcinogenesis protocol will be applied to such mice and tumor cell proliferation, migration and invasion will be determined. PACKAGE A2 3 State of the Art 3.1 State of knowledge in the field Non-melanoma skin cancer, such as basal cell carcinoma (BCC) and squamous cell carcinoma (SCC) is a very common malignancy. SCC as a solid tumour is composed of transformed epidermal keratinocytes with a highly invasive growth and tendency to metastasize. Both in vitro and in vivo model systems demonstrated that malignant transfor-mation of epidermal cells is a multistage process, in which stepwise accumulation of genetic and epigenetic events determines the transition from normal to malignant cellular state. However, the onset and the order of genetic alterations that lead to development of most sporadic cancers remain undefined. Mouse skin carcinogenesis has been an important tool for developing the current concepts regarding human neoplasia and the multistage nature of tumour development and progression. In fact, some types of mutation in oncogenes and tumour suppressor genes identified in mouse skin models also occur in human epithelial cancers. One of the best-defined experimental in vivo systems for epithelial cancer develop-ment is the chemically induced tumour model of mouse back skin. Treatment of the skin with the carcinogen 7,12-dimethylbenz-[a]-anthracene (DMBA) and the tumour promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) result in the formation of benign papillomas (PAPs) and malignant tumours (SCCs). Using this model, the timing of genetic and chromosomal alterations, as well as the cellular crosstalk between epithelial cells and cells of the microenvironment (e.g. fibroblasts, endothelial cells and immune cells) that take place during the different stages of tumour development and progression can be studied. 3.2 Previous work by the participants Using the DMBA/TPA model the central contribution of signal transduction pathways funnelling into transcription factor AP-1 (Fos/Jun) to premalignant conversion and malignant progression of epidermal cells was described. Using gene expression profiling from specimens of TPA-treated back skin and benign and malignant tumours derived from the DMBA/ TPA model we have identified novel TPA-inducible genes in mouse skin including the mucin-like glycoprotein podoplanin (PDPN). We identified pdpn as a direct c-Fos target gene being part of a Fosdependent genetic program in both the DMBA/TPA and the genetic K5-SOS-F transgenic skin 16 tumor model exhibiting expression in tumor cells, particularly at the tumor-stroma border. Despite the i) correlation between pdpn expression and tumor cell invasion, malignant progression and metastasis in mouse tumor models, as well as poor prognosis and metastatic risk in human cancer, and ii) accelerated cell motility and invasion in vitro and induced tumor growth in a xenograft model upon ectopic Pdpn, the role of PDPN in SCC formation and progression have not been addressed. 4.2 Experimental program 1. We will use well-known HaCaT cell lines harbouring additional Ras mutations, which exhibit high invasive capacity in vitro in 3D skin equivalent models (organotypic cultures). We will use this system to introduce both a pdpn siRNA producing lentiviral vector (available in the lab) and CRISPR/Cas9-mediated mutagenesis leading to significant reduction and complete loss of PDPN expression, respectively, in the tumor cells. PDPN compromised cells will be analyzed for i) cell proliferation, ii) (trans) migration and iii) invasion through matrigel matrix and in 3D organotypic cultures 2. We have recently generated floxed pdpn mice, in which we already confirmed efficient deletion of pdpn sequences via Cre recombinase technology. Crossing these mice with K14-cre mice, applying full thickness wound healing conditions (which provoke massive expression of pdpn in epithelial cells at the leading edge) Pdpn expression is completely abolished in vivo in keratinocytes. In the present project, we will apply short-term TPA treatment on mouse skin (known to strongly induce pdpn expression in basal layer keratinocytes) to evaluate the role of Pdpn in skin hyperplasia. In addition, we will apply the DMBA/TPA protocol of chronic TPA treatment to induce papillomas and subsequently SCC in WT and PDPN KO mice. Both tumor incidence and tumor volume will be determined. Tumors will be harvested and characterized by indicative immuno-histochemical analysis to define the nature of the tumor cells (particularly at the tumor-stroma border) including their ability to execute EMT. 4.3 Collaborations with other Projects in the RTG We will provide expertise of the chemical- induced skin carcinogenesis model to projects 3 and 9 and collaborate with both projects on transcriptional control of PDPN by the crucial SCC regulator Wnt (project 3) and by Ripoptosome-associated cell death pathways (project 9). 5 References 1. Hummerich L, Müller R, Hess J, Kokocinski F, Hahn M, Fürstenberger G, Mauch C, Lichter P, Angel P. 2006. Identification of novel tumour-associated genes differentially expressed in the process of squamous cell cancer development. Oncogene 25:111-21 2. Wicki A, Christofori G. 2007. The potential role of podoplanin in tumour invasion. Br J Cancer 96:1-5. 3. Gebhardt C, Riehl A, Durchdewald M, Németh J, Fürstenberger G, Müller-Decker K, Enk A, Arnold B, Bierhaus A, Nawroth PP, Hess J, Angel P. 2008. RAGE signaling sustains inflammation and promotes tumor development; J Exp Med 205:275-85 4. Peterziel H, Müller J, Danner A, Barbus S, Liu HK, Radlwimmer B, Pietsch T, Lichter P, Schütz G, Hess J, Angel P. 2012 Expression of podoplanin in human astrocytic brain tumors is controlled by the PI3K-AKT-AP-1 signaling pathway and promoter methylation. Neuro Oncol 14:426-39 5. Durchdewald M, Guinea-Viniegra J, Haag D, Riehl A, Lichter P, Hahn M, Wagner EF, Angel P*, Hess.J. 2008. Podoplanin is a novel Fos target gene in skin carcinogenesis. Cancer Res 68:6877-83 * corresponding author 17 PACKAGE A2 4 Project Plan 4.1 Specific Aims This project will apply a loss-of-function approach to define the function of PDPN in SCC formation and progression in vitro and in vivo. • apply siRNA and CRISPR/Cas9 technologies to abolish PDPN expression in SCC cell lines to measure cell proliferation, migration and invasion • generate mice lacking PDPN expression in keratinocytes to define the impact of pdpn deletion on skin homeostasis, hyperplasia and tumor development and progression Project 6: Crosstalk between melanoma cells and the blood-brain barrier: impact on coagulation and brain metastasis to identify new anti-metastatic targets. Principal Investigator: Prof. Dr. Stefan W. Schneider; Section of Experimental Dermatology, Dept. of Dermatology Mannheim, Prof. Dr. Frank Winkler; Dept. of Neuro-Oncology Heidelberg; Heidelberg University London Project Partner: Prof. Anthony Dorling, King`s College London Short Summary Human malignant melanoma is a highly metastatic tumor, and especially metastatic lesions in the brain are associated with poor prognosis. To metastasize to the brain, cancer cells must interact with cerebral endothelial cells (ECs) and migrate through the blood-brain barrier (BBB). The vascular endothelium is activated by tumor cells, which is followed by the release of inflammatory cytokines and the procoagulatory protein von Willebrand factor (VWF) known to promote tumor progression. Although treatment with heparin, a known anti-coagulant, revealed a therapeutic effect in experimental models, the underlying mechanisms are poorly understood. To investigate the impact of the coagulation system on tumor spreading, we will analyze the molecular pathways of melanoma-induced EC activation using an in vitro model of the BBB, and we will address the effect of melanoma-derived factors on EC permeability. Furthermore, taking advantage of a novel in vivo multiphoton microscopy model allowing real-time imaging of brain metastases, we will investigate how the coagulation pathway influences melanoma cell arrest and extravasation. Finally, we will characterize the ret-transgenic mouse melanoma model to evaluate therapeutic effects of anti-coagulants in vivo. PACKAGE A2 3 State of the Art 3.1 State of knowledge in the field Melanoma has the highest propensity to metastasize to the brain, and brain metastases are a major cause of mortality. Although little is known about the interaction between melanoma cells and brain microvascular endothelial cells (BMECs), malignant cells need to overcome the BBB to form brain metastasis. We hypothesize that this interaction is a multimodal process that includes melanoma cell-induced EC activation followed by the development of a proinflammatory and procoagulatory EC surface that facilitates melanoma cell adhesion. The bidirectional melanomaEC interaction leads to an increase in BBB permeability followed by melanoma cell transmigration and metastases formation. This hypothesis is supported by clinical and experimental reports showing that tumor-mediated activation of the coagulation system enhances the risk of thromboembolism and promotes tumor cell spreading in patients. Patients treated with heparins showed a better outcome and heparins reduced the formation of metastasis in animal models. 3.2 Preliminary work by the participants In previous studies we could show that melanoma-derived MMP-1 activates ECs followed by the release of proinflammatory and procoagulatory factors. Recently, we described two additional pathways that enable melanoma cells to stimulate ECs. First, a tissue factor (TF)-thrombin-PAR1 dependent pathway was discovered. Second, we identified melanoma-derived VEGF acting via VEGF-R2 as the main direct activator of ECs. This melanoma-induced EC activation was attenuated by heparins. All these pathways induce an acute Weibel-Palade body (WPB) exocytosis and the formation of ultralarge VWF fibers at the luminal surface of ECs, directly mediating platelet adhesion. Moreover, we could show that VWF supports leucocyte extravasation by increasing endothelial permeability, a process that may exhibit similarities with tumor cell extravasation. Moreover, our data show that melanoma cell-induced EC activation depends on the type of melanoma cells and ECs. However, studies on the molecular mechanisms of melanoma cell interaction with the endothelium of the BBB are lacking. The Winkler lab has established novel applications of in vivo multiphoton microscopy (MPLSM), where brain endothelial cells, blood perfusion, single cancer cells in subcellular resolution, and the single steps of brain metastasis formation of melanoma cells can be imaged through a cranial window in real time over months. Application of this technology allows the study the interaction of melanoma cells with brain ECs in 18 the physiological microenvironment. This unique experimental platform is available to other projects of this proposal to investigate the role of molecular pathways and the effect of therapeutic intervention on distinct steps of the metastatic cascade. All in all, new mechanistic insights into the crosstalk between melanoma and BMECs may have important consequences for diagnostic and therapeutic strategies in patients suffering from malignant melanoma. 4 Project Plan 4.1 Specific Aims 1. Impact of coagulation on tumor spreading 2. Mechanisms of melanoma - blood-brain barrier interaction 3. Impact of anticoagulants on brain metastasis in an animal model 4.3 Collaborations with other Projects in the RTG Cell-based assays and in-vivo stemness reporter systems to identify the role of melanoma stem cells will be performed with Projects 1 and 2. Angiopoetin-2 and its impact on BBB permeability will be analyzed with Project 8. Analysis of brain metastases in the ret mouse model will be characterized together with Project 13. 5 1. 2. 3. 4. 5. References Desch A, Strozyk EA, Bauer AT, Huck V, Niemeyer V, Wieland T, Schneider SW. 2012. Highly Invasive Melanoma Cells Activate the Vascular Endothelium via an MMP-2/Integrin alphavbeta5-Induced Secretion of VEGF-A. Am J Pathol 181:693-705. Kerk N, Strozyk EA, Poppelmann B, Schneider SW. 2010. The mechanism of melanoma-associated thrombin activity and von Willebrand factor release from endothelial cells. J Invest Dermatol 130:2259-2268. Schneider SW, Nuschele S, Wixforth A, Gorzelanny C, Alexander-Katz A, Netz RR, Schneider MF. 2007. Shearinduced unfolding triggers adhesion of von Willebrand factor fibers. Proc Natl Acad Sci USA 104:7899-7903. Pappelbaum KI, Gorzelanny C, Grässle S, Suckau J. Laschke MW, Bischoff M, Bauer C, Schorpp-Kistner M, Weidenmaier C, Schneppenheim R, Obser T, Sinha B, Schneider SW. 2013. Ultra-large von Willebrand factor fibers mediate luminal Staphylococcus aureus adhesion to an intact endothelial cell layer under shear stress. Circulation 128:50-59. Kienast Y, von Baumgarten L, Fuhrmann M, Klinkert WE, Goldbrunner R, Herms J, Winkler F. 2010. Real-time imaging reveals the single steps of brain metastasis formation. Nat Med 16:116-122. 19 PACKAGE A2 4.2 Experimental program 1. The molecular mechanisms of melanoma-mediated activation of BMECs in vitro and on the integrity of the EC monolayer will be analyzed. In previous work we have found that some melanoma cell lines such as A2058 can form parenchymal brain metastases, whereas other lines (i.e. B16F10) do not. The metastatic potential is reflected by a distinct ability of EC activation. In order to clarify the molecular mechanisms of melanoma-derived mediators that activate ECs, different melanoma cell lines will be compared by gene expression analysis and proteome profiling. 2. a) The impact of melanoma-mediated EC activation on expression of adhesion molecules and melanoma cell adhesion will be assessed using microfluidic devices established by the Schneider lab. b) In vivo MPLSM imaging of VWF and adhesion molecules, and their colocalization with arrest and extravasation of melanoma cells in the brain will be investigated. 3. Finally, we will study the relevance of our results using the ret-transgenic mouse melanoma model. To this end, VWF-selective changes in BBB integrity will be evaluated by analysis of candidate molecules for vascular permeability, by application of heparins, new anticoagulants and knockdown of adhesion factors in melanoma cells in our in vivo MPLSM imaging model. We will thereby determine their role in the metastatic cascade, which should lead to rapid identification of most promising therapeutic targets. Project 7: Liver-specific endothelial mechanisms of melanoma metastasis Principal Investigator: PD Dr. Cyrill Géraud, Prof. Dr. Sergij Goerdt, Dept. of Dermatology, Medical Faculty Mannheim, Heidelberg University London Project Partner: Dr. Ilaria Malanchi, Tumor Host Interaction Group, London Research Institute, Cancer Research UK, London Short Summary Hematogenous metastasis is remarkably organ-specific. The liver with its unique sinusoidal vascular system is one of the preferred sites of malignant melanoma (MM) metastasis. Liverspecific, endothelial-dependent mechanisms of MM cell dissemination will be analyzed by scrutinizing liver sinusoidal endothelial cell (LSEC)-specific candidate molecules such as Stabilin-1 and Stabilin-2 identified by us as well as other scavenger and lectin-like receptors. Their role in tumor cell adhesion and transmigration will be investigated in a microfluidic chamber model in vitro using over-expression in human umbilical vein endothelial cells (HUVEC) as well as by using LSEC from the respective KO animals. A special focus will be on hyaluronan (HA)-mediated interactions. Results will be confirmed in vivo using the ret model as well as a B16 luciferase model of MM metastasis to the liver in WT and KO animals. The final goal of this project is to develop novel strategies to treat metastasis in this devastating disease. PACKAGE A2 3 State of the Art 3.1 State of knowledge in the field Metastatic spread to distant organs in general comprises a series of steps in which endothelial cells (EC) are intricately involved such as tumor cell adhesion and transmigration. In many cancers, including MM, however, cancer cell dissemination is remarkably organ-specific. Organspecific metastasis is likely caused by tumor cell heterogeneity as well as by organ-specific stromal factors. Among these, EC heterogeneity may impact on tumor-EC interactions by modulating the well-known general adhesive mechanisms or by providing organ-specific pathways. Besides the lungs and the brain, the liver is a preferred site for distant metastasis in MM, and the primary site for metastasis in uveal melanoma. LSEC are a prime example of organ-specific EC differentiation. LSEC selectively express several scavenger and lectin-like receptors such as stabilin-1/2, LYVE-1, MRC1, CD32B, CLEC-1B, -4G, and -4M. Two of these molecules, stabilin-2 and LYVE-1 are known HA receptors. In addition, the HA receptor CD44 is also expressed by LSEC. Furthermore, HA-dependent mechanisms have been shown to be of high importance for melanoma metastasis in general and especially in the liver. HA is generated by HA synthases on the surface of MM cells and could contribute to liver metastasis by binding to stabilin-2, LYVE-1 or to CD44. Conversely, CD44 has also been found on many types of tumor cells including MM. Therefore, MM cells could also adhere to HA bound by the three HA receptors on the surface of LSEC. In addition, stabilin-1 and Clec-4G/LSECtin have already been demonstrated to mediate binding of tumor cells other than MM cells to LSEC. In summary, the analysis of this set of candidate LSEC adhesion molecules may open new avenues to target EC-dependent, organ-specific MM metastasis to the liver. 3.2 Previous work by the participants Our group has a longstanding track record in analyzing the specific molecular repertoire and functions of LSEC including identification of the scavenger receptors stabilin-1 and -2. Stabilin-2 KO mice, although displaying no obvious phenotype, show highly increased plasma levels of HA proving that stabilin-2 is the major receptor for HA turnover. Due to impaired clearance of other noxious blood factors, stabilin-1/2-/- double deficient mice have a reduced lifespan and develop severe glomerulosclerosis indicating the importance of stabilin function for the whole organism. LSEC specifically produce wnt2 that acts as an autocrine growth factor by cross-stimulating the VEGF pathway. Comprehensive gene expression analysis revealed a LSEC-specific, hepatic microenvironment-dependent differentiation program comprising distinct sets of growth and transcription factors as well as of adhesion- and endocytosis-associated molecules including the novel junctional protein Leda-1. 20 4 Project Plan 4.1 Specific Aims The general aim of the project is to analyze the organ-specific, endothelial-dependent mechanisms of MM cell dissemination to the liver. For this purpose, we will thoroughly study MM-LSEC interactions in vitro and in vivo. In vitro, we will analyze (1) MM cell-LSEC adhesion and transmigration in a microfluidic chamber model using HUVEC retrovirally transfected with LSECspecific candidate adhesion molecules, as well as LSEC isolated from the respective knockout mice. Special attention will be given to HA-mediated mechanisms. In vivo, (2) LSEC-dependent MM metastasis to the liver will be investigated using a B16 luciferase model, the ret MM model, and a human xenotransplant model of MM metastasis. Experimental program 1. LSEC-specific candidate molecules for liver-specific melanoma-EC adhesion and transmigration, i.e. stab1, stab2, Lyve-1, MRC1, CD32b, CLEC-1B, -4G, and -4M, will be retrovirally transfected into HUVEC. Using transwell migration assays and a microfluidic chamber device that simulates organ-specific flow conditions, MM cell adhesion and transmigration will be studied using transfected HUVEC as well as murine LSEC and – as a control – murine lung microvascular endothelial cells (LMEC) from wild-type and knock out animals (stab1-/-, stab2-/-, Lyve-1-/-, CD44-/-). In these assays, the relevance of HAdependent mechanisms will be analysed by pre-incubation of either MM or endothelial cells with HA to block HA binding proteins, by hyaluronidase treatment and by inhibition of HA synthases. The function of CD44 expressed by MM cells as a ligand for HA deposited on LSEC will be investigated by CD44 knock-down in MM cells. 2. In vivo, organ-specific MM cell-EC adhesion and metastasis will be analysed by injecting luciferase-expressing B16F10 mouse MM cells into the spleen (liver metastasis) or tail vein (lung metastasis) of wild-type and knockout animals (stab1-/-, stab2-/-, Lyve-1-/-, CD44-/-). HA dependent mechanisms will be studied in vivo using i.p. injection of hyaluronidase and p.o. administration of HA synthase inhibitors. Development of metastases will be traced and quantified by bioluminescence in vivo imaging. Adhesion molecule-deficient animals will be back-crossed with ret MM mice and analysed for spontaneous liver metastases. Adhesion molecules that can be shown to be involved in MM adhesion and metastasis in those murine models will be further scrutinized in a xeno-transplant model of human melanoma metastasis. 4.3 Collaborations with other Projects in the RTG Melanoma cell-LSEC adhesion will be analyzed with Project 6 (microfluidic chamber) and Project 1 (CD44-mediated adhesion). The ret model of MM and the human MM mouse xeno-transplant model will be provided by Projects 13 and 12, respectively. 5 1. 2. 3. References Schledzewski K*, Geraud C*, Arnold B, Wang S, Grone HJ, Kempf T, Wollert KC, Straub BK, Schirmacher P, Demory A, Schonhaber H, Gratchev A, Dietz L, Thierse HJ, Kzhyshkowska J, Goerdt S. 2011. Deficiency of liver sinusoidal scavenger receptors stabilin-1 and -2 in mice causes glomerulofibrotic nephropathy via impaired hepatic clearance of noxious blood factors. J Clin Invest 121:703-14. Geraud C*, Schledzewski K*, Demory A, Klein D, Kaus M, Peyre F, Sticht C, Evdokimov K, Lu S, Schmieder A, Goerdt S. 2010. Liver sinusoidal endothelium: a microenvironment-dependent differentiation program in rat including the novel junctional protein liver endothelial differentiation-associated protein-1. Hepatology 52:313-26. Klein D, Demory A, Peyre F, Kroll J, Augustin HG, Helfrich W, Kzhyshkowska J, Schledzewski K, Arnold B, Goerdt S. 2008. Wnt2 acts as a cell type-specific, autocrine growth factor in rat hepatic sinusoidal endothelial cells crossstimulating the VEGF pathway. Hepatology 47:1018-31. 21 PACKAGE A2 4.2 Research Area B – Primary Resistance to Cell Death and Immunity Project Package B1: Primary Resistance to Cell Death (Projects 8-10) Project 8: Does Angiopoietin-2 protect malignant melanoma tumor cells from anoikis? Principal Investigator: Dr. Moritz Felcht, Dept. Dermatology, Med. Faculty Mannheim, Heidelberg University; Prof. Dr. Hellmut G. Augustin, Vascular Biology & Tumor Angiogenesis, Med. Faculty Mannheim, Heidelberg University, and German Cancer Research Center, Heidelberg London Project Partner: Prof. Kairbaan Hodivala-Dilke, Centre for Tumor Biology, Barts Cancer Institute, Barts and The London, Queen Mary University College of London Short Summary Increased levels of the Tie2 ligand Angiopoietin-2 (Ang-2) can be detected in the blood from patients suffering from metastasized malignant melanoma (MM) (AJCC III/ IV). Furthermore, different studies could show that Ang-2 is essential for primary MM tumor growth and metastasis formation in mice. Ang-2 is mainly secreted by activated endothelial cells but can also be produced by MM tumor cells themselves. Therefore, Ang-2 may act in an autocrine as well as in a paracrine manner on MM tumor cells. In the absence of its high affinity receptor Tie2, Ang-2 directly associates with and signals through integrins. Integrin activation is required during metastasis formation to protect tumor cells from anoikis, apoptosis induced by inadequate cell-matrix connection. Preliminary data by the applicants show that Tie2 negative MM tumor cells are protected from anoikis by exogenous Ang-2 stimulation. Consequently, this project aims to study if anoikis resistance mediated by Ang-2 represents an essential new pathomechanism during MM metastasis formation and if this can be used therapeutically. PACKAGE B1 3 State of the Art 3.1 State of knowledge in the field: Angiopoietin-2 (Ang-2) is essential during MM metastasis formation and increased levels of Ang-2 can be detected when metastases have been formed. Metastasis formation passes through the single steps invasion, intravasation, intravascular survival, extravasation and colony formation. Intravascular survival of tumor cells requires protection from anoikis, apoptosis induced by inadequate cell-matrix connection. Protection from anoikis is classically acquired by integrin activation [3] but may also be achieved by receptor activation. Recently, the applicants could show that integrins may also be activated by Ang-2 in the absence of Tie2 receptor. Ang-2 can be detected in some Tie2 negative malignant melanoma cell and may therefore directly bind to and activate integrins. Yet, the impact of Ang-2 stimulation of MM cells has not been studied. 3.2 Previous work by the participants: In the presence of Tie2, Ang-2 induces complex formation of Tie2-FAK-αvβ3 integrin, FAK phosphorylation at Ser910 but not at Tyrosine397, induces αvβ3 integrin internalization/ degradation and endothelial destabilization (Thomas*, Felcht*, et al, 2010). In the absence of Tie2 receptor Ang-2 binds αvβ3, αvβ5 and α5β1 integrins and induces FAK phosphorylation at Tyrosine397 and RAC activation (Felcht et al., 2012). The binding of Ang-2 to integrins is tightly regulated and obligates absence of the high affinity receptor Tie2, an acid environment and integrin expression in their active conformation (Felcht et al., 2012). Functionally, Ang-2 induces in endothelial cells migration and sprouting independent of Tie2 expression (Felcht et al., 2012). In A375 MM tumor cells Ang-2 stimulation protects from anoikis in vitro (unpublished). A375 cells express αvβ3 integrin but not Tie2 receptor or αvβ5 (unpublished). Tumor specimens of metastatic MM show CD34 negative/ αvβ5 expressing and/or αvβ3 expressing cells (unpublished). 4 Project Plan 4.1 Specific Aims I. Which requirements are needed for Ang-2 protection from anoikis? II. Is there a therapeutic relevance of Ang-2 induced protection from anoikis? 22 4.3 Collaborations with other Projects in the RTG Primary cutaneous MM cells will be generated in collaboration with project 2. Vascular remodelling/pruning is studied in collaboration with project 3. The studies of molecular signalling of apoptosis will be supported by project 9 and 10. The ret transgenic melanoma mouse model will be provided by project 13. 5 1. 2. 3. 4. 5. References Felcht M, Luck R, Schering A, Seidel P, Srivastava K, Hu J, Bartol A, Kienast Y, Vettel C, Loos EK, Kutschera S, Bartels S, Appak S, Besemfelder E, Terhardt D, Chavakis E, Wieland T, Klein C, Thomas M, Uemura A, Goerdt S, Augustin HG. 2012. Angiopoietin-2 differentially regulates angiogenesis through TIE2 and integrin signaling. J Clin Invest 122:1991-2005. Thomas M*, Felcht M*, Kruse K, Kretschmer S, Deppermann C, Benest AV, Fiedler U, Augustin HG. 2010. Angiopoietin-2 stimulation of endothelial cells induces alphavbeta3 integrin internalization and degradation. J Biol Chem 285:23842-9. *equal contribution Augustin HG, Koh GY, Thurston G, Alitalo K. 2009. Control of vascular morphogenesis and homeostasis through the angiopoietin-Tie system. Nat Rev Mol Cell Biol 10:165-177. Helfrich I, Edler L, Sucker A, Thomas M, Christian S, Schadendorf D, Augustin HG. 2009. Angiopoietin-2 levels are associated with disease progression in metastatic malignant melanoma. Clin Cancer Res 15:1384-92. Nasarre P, Thomas M, Kruse K, Helfrich I, Wolter V, Deppermann C, Schadendorf D, Thurston G, Fiedler U, Augustin HG. 2009. Host-derived angiopoietin-2 affects early stages of tumor development and vessel maturation but is dispensable for later stages of tumor growth. Cancer Res 69:1324-33. 23 PACKAGE B1 4.2 Experimental program (I.A.) Various MM tumor cells (C32, SK-Mel-28, RPMI 7951, HAT 144, SK-Mel2, WM9, WM35, MV3, CRL 1676, Malme 3M, MeWo, WM 1158) will be compared for their Ang-2 induced anoikis resistance in different anoikis assays. These studies will include primary MM cells as well as Ang-2 studies in the ret transgenic melanoma mouse model. PCR/ELISA studies will compare Ang-2 levels. (I.B.) Integrin expression of the MM cells (see I.A.) will be compared by PCR, Western Blot, IP and FACS analyses i. Inhibition studies (antibodies, siRNA, shRNA) should unravel the relevance for anoikis resistance. (I.C.) Tie2 expression will be studied in the different MM cells (see I.A.) (PCR/Western blot). Inhibitory (siRNA, antibodies)/overexpression (shRNA) studies will be compared with the integrin expression/anoikis sensitivity. (I.D.) Intra- and extracellular Ang-2 signalling has been observed. MM cells with low, intermediate & high levels of Ang-2 (see I.A.) will be analysed in inhibition studies (siRNA, shRNA, antibody) in the anoikis assay. Exogenous (recombinant, conditional media w. adenoviral overexpression [Ad-Ang-2])) vs. endogenous (Ad-Ang-2) Ang-2 stimulation will support the analyses. (I.E.) AKT, ERK, mTOR, JNK, Mcl-1 and bad signalling will be analysed. Intrinsic/extrinsic apoptosis will be studied in collaboration with project 9. Pharmacological inhibitory studies (Worthmanin, UO126, Rapamycin, zVAD-fmk) will be performed in parallel. (I.F.) MM cells (different integrin/Ang-2/ Tie2 expression profiles) will be used for metastasis studies in vivo. Metastasis formation will be studied by conventional microscopy and correlated with anoikis sensitivity. Control experiments with Ang-2 overexpression and inhibition (antibodies) will support the in vivo study. Inhibition experiments (antibodies) within the ret transgenic melanoma mouse model will finalize the in vivo studies. (II.A.) Preliminary studies detected αvβ3 integrin in nonvascular MM areas in tumor specimens from patients. Integrin expression studies (see I.B.) will be performed in a larger cohort with co-staining against melanocytic markers. (II.B.) The signalling studies (see I.e.) will be followed by combination inhibitory studies in vitro. Project 9: The regulation of Ripoptosome-associated cell death pathways in keratinocyte skin cancer Principal Investigator: Prof. Dr. Martin Leverkus, Section of Molecular Dermatology, Department of Dermatology; Medical Faculty Mannheim, Heidelberg University London Project Partner: Prof. Pascal Meier, ICR, Head of Apoptosis Team, The Break-through Toby Robins Breast Cancer Research Centre, Institute of Cancer Research, London Short Summary For progression and metastasis of SCC, the crosstalk of transformed keratinocytes with tumor stroma and immune cells is of importance. Cell death resistance is a prerequisite for progression and metastasis. We recently showed that the Ripoptosome – an intracellular signalling platform containing RIP1, Caspase 8, FADD, and cFLIP - controls apoptosis and necroptosis in SCC. The Ripoptosome is necessary for signaling initiated by membrane-bound receptors (death receptors, TLR 3) and thus shapes the quality not only of cell death but also of the immune response potentially activated by necroptosis. The project will investigate the structure, function, and assembly of the Ripoptosome in different progression stages of primary and transformed keratinocytes and SCC in vitro and in vivo. We will use in vitro model systems (HaCaT tumor progression model) and representative tumor cell lines in vitro. Functional studies will make use of lentiviral knockdown or overexpression of constituents of the Ripoptosome depending on their expression. Our project promotes understanding of the function of different components of the Ripoptosome for tumor progression and metastasis of SCC. PACKAGE B1 3 State of the Art 3.1 State of knowledge in the field The major problem of SCC is its multiplicity, whereas metastasis only occurs in locally progressed stages. The reason for this relative resistance to metastasis is currently unclear but may involve immune surveillance by the host activated by inflammatory signals. Cellular death and inflammatory pathways in SCC are regulated by membrane bound receptors that can activate caspases in a pro-apoptotic or inflammatory manner. Activation of cell death via TLR7 by its ligand imiquimod is sufficient for elimination of a high proportion of in situ carcinoma by TRAIL-dependent cell death. Cell death activation can occur via the so-called Death Inducing Signaling Complex (DISC) for death receptors (DR). Alternatively cell death is initiated by recruitment of adaptor molecules such as TRIF or MyD88 to TLR3 or other TLRs. Most of these signaling platforms mediate apoptosis via FADD and caspase 8, whereas more recently the role of the RIP1-RIP3 module (necrosome) for receptor-induced necroptosis has been demonstrated. The decision for the outcome of receptor activation is dependent on the regulation of the components and activity within the Ripoptosome, a recently described novel intracellular signaling platform. The inhibitor of apoptosis proteins (IAPs) such as cIAP1/2 and XIAP were shown to suppress the formation of some of these signaling platforms, in particular the Ripoptosome. The importance of the Ripoptosome for receptor-induced cell death responses in different progression stages of SCC is unknown to date and will be investigated in this project. 3.2 Preliminary work by the participants In our preliminary work we have studied the formation of the molecular complex that we named the Ripoptosome, which consists of FADD, RIP1, caspase 8 and cFLIP. Our studies were performed in HaCaT and in advanced tumor progression forms derived from HaCaT. By studying these cellular models we could demonstrate the decisive role of RIP1 for the formation of the Ripoptosome, and a crucial role of the RIP1 kinase activity for necroptosis execution induced by death receptors or TLR3 ligands. Furthermore, we could identify the critical role of cFLIP in promoting cell death resistance of A5RT3 (the metastatic cell line of the HaCaT tumor progression model) to cell death mediated by the Ripoptosome. We will therefore aim to further study assembly, activation, and execution of cell death by the Ripoptosome, a complex known to mediate both apoptosis and 24 necroptosis. Ultimately we will aim to investigate if modulation of this complex can be exploited as a novel target for tumor therapy of early or late SCC of the skin. 4.2 Experimental program 1. First we will analyze the differences in expression of key complex components (FADD, RIP1, RIP3, initiator caspases 8 and 10, Mixed lineage kinase domain-like (MLKL)) in various cell lines at the mRNA (qPCR) and protein levels (Western blotting). To this end we will use a number of model cell lines (HaCaT and MET SCC progression models) as well as primary keratinocytes and SCC cells and compare results with data derived from ex vivo analysis of human tumors (immunohistochemistry, qPCR, Western blotting). These studies will help to define the main differences between primary keratinocytes and SCC cells. 2. The next step will be the functional analysis of cell death in the above-mentioned primary cells and cell lines. We will analyze the caspase- and RIP1 kinase-dependency of cell death in response to a number of stimuli (TLR or death ligands) by using inhibitors (zVAD-fmk, Necrostatin-1, necrosulfonamide). The morphology of cell death will be determined with combinations of Hoechst and SYTOX Green staining in a kinetic manner using fluorescent microscopy. Caspase-dependent cell death will be monitored by initiator/effector caspase cleavage. Using reverse transfection, transient siRNA transfection against RIP3, MLKL, or caspase-8 will be utilized to dissect which signalling pathways are required for cell death execution. 3. We will then study Ripoptosome formation and activity in selected cell lines (primary and lines derived from SCC progression models) using caspase-8 coimmunoprecipitation. Finally we want to analyze SCC cell lines that show primary resistance to cell death stimuli. We will investigate which Ripoptosome components are critical for cell death resistance, and if proteins such as RIP1, RIP3, MLKL, or others can reconstitute or block the ability of cells to undergo Ripoptosome-induced cell death using retro/lentiviral knockdown of these key components. For selected questions we will extend these studies to organotypic model systems to investigate if identified genes shape a differential cell death response in a three-dimensional environment. 4.3 Collaborations with other Projects in the RTG Our project will utilize nude mouse model systems (Project 2) and skin carcinogenesis models (Project 5) to extend the in vitro findings in vivo. Genetic manipulation to eliminate Ripoptosomeassociated genes in SCC cell lines via CRISPR/Cas9 will be done in collaboration with project 3 and 4. 5. References 1. Panayotova-Dimitrova D, Feoktistova M, Ploesser M, Kellert B, Hupe M, Horn S, Makarov R, Jensen F, Porubsky S, Schmieder A, Zenclussen AC, Marx A, Kerstan A, Geserick P, He YW, Leverkus M. 2013. cFLIP regulates skin homeostasis and protects against TNF-induced keratinocyte apoptosis. Cell Rep 5:397-408. 2. Cullen SP, Henry CM, Kearney CJ, Logue SE, Feoktistova M, Tynan GA, Lavelle EC, Leverkus M, Martin SJ. 2013. Fas/CD95-Induced Chemokines Can Serve as "Find-Me" Signals for Apoptotic Cells. Mol Cell 17:1034-1048 3. Feoktistova M, Geserick P, Kellert B., Dimitrova DP, Langlais C, Hupe M, Cain K, MacFarlane M, Hacker G, Leverkus M. 2011. cIAPs block Ripoptosome formation, a RIP1/caspase-8 containing intracellular cell death complex differentially regulated by cFLIP isoforms. Mol Cell 43:449-463. 4. Geserick P, Hupe M, Moulin M, Wong WW, Feoktistova M, Kellert B, Gollnick H, Silke J, Leverkus M. 2009. Cellular IAPs inhibit a cryptic CD95-induced cell death by limiting RIP1 kinase recruitment. J Cell Biol 187:1037-1054. 5. Kavuri SM, Geserick P, Berg D, Dimitrova DP, Feoktistova M, Siegmund D, Gollnick H, Neumann M, Wajant H, Leverkus M. 2011. Cellular FLICE-inhibitory protein (cFLIP) isoforms block CD95 or TRAIL-induced NF-kB activation independent of caspase-8 cleavage. J Biol Chem 286:16631-16646. 25 PACKAGE B1 4 Project Plan 4.1 Specific Aims 1. Compare expression of the key Ripoptosome components in SCC tumor progression models in vitro, primary keratinocytes and SCC cell lines and primary tumor samples to identify differential expression. 2. Investigate the qualitative and quantitative cell death responses in SCC tumor progression models and compare it to primary human keratinocytes. 3. Compare Ripoptosome formation and activity in selected cell lines and investigate the functional relevance of its components by knockdown or overexpression studies. Project 10: IAP antagonists: A novel therapeutic option to overcome cell death resistance in malignant melanoma? Principal Investigator: Dr. Peter Geserick, Prof. Dr. Martin Leverkus, Dept. of Dermatology, Medical Faculty Mannheim, Heidelberg University London Project Partner: Prof. Henning Walczak, Cell Death and Inflammation Laboratory, Cancer Institute, University College London Short Summary Acquisition of cell death resistance is a critical step during skin tumorigenesis. Inhibitor of apoptosis proteins (IAPs) are negative regulators of caspase-dependent apoptotic (e.g. X-linked IAP) and receptor interacting protein 1 (RIP1)-dependent necroptotic (e.g. cIAPs) cell death induced by death receptors. The caspase-8 inhibitor cFLIP is upregulated during tumor progression in malignant melanoma, and protects tumor cells from receptor-mediated cell death. Notably, the short isoform of cFLIP (cFLIPS) protects cells from apoptotic but not necroptotic cell death. This project will study how different IAPs regulate the diverse forms of cell death in malignant melanoma. Therefore, we will investigate (1) how cIAPs regulate the quality and quantity of cell death in cultured malignant melanoma cells; (2) the function of different components of intracellular death pathways and the impact of cFLIP isoform expression on apoptotic and necroptotic cell death and; (3) the potential of IAP antagonists for tumor suppression in xenograft mouse models. These studies will functionally validate the role of cFLIP isoforms in conferring resistance to different forms of cell death, and will elucidate how this relates to the overall therapeutic response in malignant melanoma. PACKAGE B1 3 State of the Art 3.1 State of knowledge in the field Activation of death receptor (DR)-induced apoptosis transmitted by cytokines such as TRAIL and CD95L is a required mechanism for elimination of unwanted and transformed cells. This process is highly controlled by antiapoptotic proteins that control caspase activity initiated either by DRs (cFLIP inhibits Caspase 8) or intracellularly (e.g. XIAP blocks caspase-9 and 3). In contrast, cIAPs inhibit necroptosis by regulation of a RIP1-controlled signalling platform. Suppression of such cell death responses mediated by upregulation of cell death inhibitory proteins during tumor progression may confer therapeutic resistance in malignant melanoma. Melanoma cells are known to be highly resistant against death ligand-mediated cell death. Inhibition of this cell death resistance by combined activation of DRs and suppression of IAP activity using the respective antagonists could represent a promising novel anti-cancer strategy for malignant melanoma. IAP antagonists suppress the caspase-inhibitory function of XIAP and additionally promote rapid degradation of cIAP1 and 2. The consequences of these activities of IAP antagonists are either increased apoptosis induction by effector caspase-3 activity, or promotion of apoptosis or necroptosis initiated by activation of the RIP1/RIP3 signalling machinery. These interesting molecular processes could be of critical relevance to overcome cell death resistances in melanomas and therefore for tumor suppression. 3.2 Previous work by the participants Over the past years our group intensively worked on the identification of resistance mechanisms in skin tumors. We identified the critical role of cFLIP for DR-induced cell death in melanoma and the role of IAPs for regulation of necroptotic cell death responses. We further demonstrate necroptosis induction in transformed, but not primary cells when IAP function is suppressed. Analysis of these 26 signalling pathways in melanoma cells demonstrated a substantial upregulation of cFLIP and IAPs, and concomitant increased resistance to death ligand-mediated cell death in malignant melanoma cells. Intriguingly, in the presence of IAP antagonists, we were able to overcome cell death resistance and increase sensitivity to TRAIL, indicative of the indispensable role of IAPs for cell death resistance in melanoma cells. A more precise role of cFLIP in the regulation of DR-induced cell death was shown by cFLIP overexpression. In line with our previous studies, both cFLIP isoforms were able to block TRAIL-induced apoptosis in melanoma. However, only cFLIPs promotes necroptotic cell death responses in the presence of IAP antagonists (Figure 1). These data are indicative of the critical but differential role of cFLIP isoforms in cell death regulation. 4.2 Experimental program A repertoire of melanoma cells representing different tumour progression stages will be analysed for expression of cFLIP and IAP proteins (XIAP, cIAP1/2) as well as of their counteracting protein molecules (caspases, RIP1/RIP3, MLKL) using various biochemical approaches (immunoblot, qPCR). The determination of the quality and quantity of cell death initiated by treatment with TRAIL or CD95L will be analyzed using crystal violet and PI/AnnexinV assay, and fluorescence microscopy. The relevance of the respective proteins (cFLIP, IAPs) involved in cell death resistance in melanomas will be investigated by genetic manipulation of the endogenous expression levels (siRNA, lentiviral shRNA, retroviral expression of cFLIP) followed by analysis of the quality and quantity of cell death upon combined IAP antagonist and DL treatment. To investigate the role of cFLIPs and IAPs for tumor growth and cell death resistance, appropriate established and biochemically- characterized melanoma cells will be xenotransplanted into immune deficient mice. Tumor growth and metastasis as well as the effect of IAP antagonists in combination with DL (TRAIL, CD95L) for cell death sensitivity and for tumor suppression will be investigated in mice with established melanoma in vivo following DL and IAP antagonist treatment. 4.3 Collaborations with other Projects in the RTG Cooperation with project 4 will allow us to perform optimal knockdown conditions in large-scale but also in small scale for specific proteins (cFLIP, IAPs) in melanoma. Cooperation with project 2 will enable us to perform experiments with xenograft mouse models. We will share our expertise in the field of cell death signalling in vitro and in vivo with projects 3, 5, 8, and 11. 5 1. 2. 3. References Feoktistova M*, Geserick P*, Kellert B, Panayotova-Dimitrova D, Langlais C, Hupe M, Cain K, MacFarlain M, Häcker G, Leverkus M. 2011. cIAPs block Ripoptosome formation, a RIP1/caspase 8 containing intracellular cell death complex differentially regulated by cFLIP isoforms. Mol Cell 43:449-63 (* equal contribution) Geserick P, Hupe M, Moulin M, Wong WW, Feoktistova M, Kellert B, Gollnick H, Silke J, M Leverkus. 2009. Cellular IAPs inhibit a cryptic CD95-induced cell death by limiting RIP1 kinase recruitment. J Cell Biol. 187:10371054. Geserick P, Drewniok C, Hupe M, Haas TL, Diessenbacher P, Sprick MR, Schon MP, Henkler F, Gollnick H, Walczak H, Leverkus M. 2008. Suppression of cFLIP is sufficient to sensitize human melanoma cells to TRAIL or CD95L-mediated apoptosis. Oncogene 27:3211-3220. 27 PACKAGE B1 4 Project Plan 4.1 Specific Aims The general aim of the project is to study the mechanistic relevance of IAPs and cFLIPs for resistance of DR-induced cell death in melanoma cells and the role of IAP antagonists as a potential therapeutic for melanoma treatment. For this purpose, we will analyze expression of IAPs and cFLIP isoforms and check the sensitivity of cells to TRAIL in the presence or absence of IAP antagonists from in a set of melanomas representing different tumour stages, and in melanoma genetically modified with downregulation or overexpression of IAPs and cFLIPs. The growth and metastasis of melanomas expressing cFLIP isoforms upon xenotransplantation into immune deficient nude mice will be assessed. Finally, the ability of a combination therapy of IAP antagonist/DL to treat established tumors in a xenograft mouse model will be analyzed. Research Area B – Primary Resistance to Cell Death and Immunity Project Package B2: Primary Resistance to Tumor Immunity (Projects 11-13) Project 11: Characterization and modulation of CC-chemokine receptor 6 (CCR6) mediated immunosurveillance in malignant melanoma Principal Investigator: Dr. Anke S. Lonsdorf, Prof. Dr. Alexander H. Enk, Department of Dermatology; Medical Faculty Heidelberg, Ruprecht-Karls-University Heidelberg London Project Partner: Prof. Fran Balkwill, Centre for Cancer and Inflammation, Barts Cancer Institute, London Short Summary The general objective of this research proposal is to evaluate the distinct functional contribution of CC-chemokine receptor (CCR) 6 interactions with its cognate ligand CCL20 in modulating specific cellular anti-melanoma immune responses. The relevance of the CCR6/CCL20 axis for the immune control of melanoma at the primary tumor site and the skin draining lymph nodes (LNs) will be studied in two established murine models: 1) the B16 transplanted melanoma model in C57BL/6 (WT) and CCR6 knockout (KO) mice and 2) the spontaneous melanoma model in Ret-transgenic mice as well as human melanoma tissue samples. An increased understanding of the functional interplay between CCR6/CCL20-guided pathways in the local control of melanoma and draining LNs may allow for the identification of novel therapeutic strategies on a molecular level. PACKAGE B2 3 State of the Art 3.1 State of knowledge in the field Chemokines, a family of small, secreted molecules, and their cognate G-protein-coupled receptors play an essential role in the elicitation of specific immune responses, particularily directed compartment-specific migration of immune and tumor cells (i.e. chemotaxis). CCL20 and the antimicrobial peptide ß-defensin expressed in the epidermis are a potent impetus for the recruitment of subsets of dendritic cell (DC), B-cells and memory T cell subsets expressing CCR6, its exclusive receptor. In addition to its constitutive expression the epidermis, CCL20 and a CCR6- expressing immune cell infiltrate has been detected in several malignancies, including melanoma. Yet, the functional contribution of the CCR6/CCL20 axis in the immune control of melanoma remains controversial: While CCR6/ CCL20 interactions have been found to support anti-melanoma immune responses in a murine model of lung metastasis, tumor-derived CCL20 has been reported to promote tumor growth and immune escape; partially by recruiting subsets of tolerogenic immature DC or regulatory T cells. Analysis of the kinetics and distribution of CCR6-guided immune cell subsets and their functional contribution for the immune control of melanoma comprises the focus of this research proposal. Furthermore, stimuli by which the expression of CCR6 ligands may be modulated at the tumor site and/ or the skin-draining LN are poorly understood and their potential relevance for anti-tumoral immune responses warrants further investigation. 3.2 Preliminary work by the participants Our group has a long-standing interest in studying mechanisms of compartment-specific trafficking of melanoma and immune cells with a particular focus on the role of chemokine interactions. In previous studies we provided evidence that CCR6 expression on DC subsets and effector T cells vitally contributes to the elicitation of skin modulating immune responses by directing their recruitment to sites of enhanced CCL20 expression (Hedrick and Lonsdorf et al., 2009). We have also demonstrated that small-molecule activators, such as toll-like receptor (TLR)-activating microbial products, support the formation of protective antitumoral immune responses in the skin (Lonsdorf et al., 2003) and that the accumulation of both, epidermal chemokines and CCR-bearing immune cells in skin-draining LNs may be amplified by topically applied immunmodulators in vivo (Chien et al., 2009; Huang et al., 2008). 28 4.2 Experimental program 1. A) Luciferase-transduced murine B16 melanoma (B16-luc) will be injected s.c. into syngeneic WT and CCR6 KO mice. Kinetics of skin tumors and LN metastases will be monitored in vivo (calliper, bioluminescence analysis). Additionally, tumor burden and melanoma metastasis will be quantified ex vivo by exploiting a bioluminescence reporter system. .Immune cell infiltrates, tumor vascularisation and the predominant chemokine/CCR and cytokine profiles will be analysed (immunohistochemistry (IHC), FACS, RT-PCR) in primary transplanted B16-luc melanomas and corresponding skin-draining LNs. Results will be validated in spontaneously arising melanomas in Ret-transgenic mice B) Subsets of CCR6-expressing immune cells from tumor-bearing WT mice will be adoptively transferred into CCR6 KO mice before/ after B16-luc inoculation. Tumor kinetics and LN metastasis, tumor-homing capabilities and alterations in the tissue microenvironment will be monitored in primary melanomas and LNs as described above and by in vivo multiphoton microscopy. 2. A) B16-luc overexpressing CCL20 (B16-luc-L20) and appropriate empty vector control cells will be injected s.c. into WT and CCR6 KO mice to study local immune cell infiltrates, tumor vascularisation and cytokine profiles (IHC, FACS RT-PCR, ELISA). Also, melanoma cells derived from B16-luc-L20 primary melanoma will be analysed for mechanisms of apoptosis resistance B) The effect of topical immunmodulators (i.e. TLR7- agonist imiquimod, DNCB) on CCL20/ ß –defensin expression, immune cell recruitment and tumor kinetics will be studied within the experimental setting and methods described above in B16-luc transplanted melanomas and melanomas of Ret-mice. Also, the effects of intratumoral injections of a blocking anti-mCCL20 antibody will be studied in both murine models. 3. Paraffin embedded human primary melanoma and LN metastases will be analyzed by IHC for CCL20 and ß -defensin expression as well as phenotype and distribution of an (CCR6expressing) immune cell infiltrate. Correlation with pathomorphological patterns, disease stage and tumor progression will be performed. 4.3 Collaborations with other Projects within the RTG Our project will utilize bioluminescence imaging (Project 7), multiphoton microscopy (Project 6) and the Ret-mouse model system (Project 13) in close collaboration. Mechanisms of apoptosis resistance in B16-luc-L20 melanoma will be studied with partners of project 10. 5 1. 2. 3. 4. 5. References Lonsdorf AS, Kraemer BF, Fahrleitner M, Schoenberger T, Gnerlich S, Ring S, Gehring S, Schneider SW, Kruhlak MJ, Meuth SG, Nieswandt B, Gawaz M, Enk AH, and HF Langer. 2012. Engagement of αIIbβ3 (GPIIb/IIIa) with ανβ3 mediates interaction of melanoma cells with platelets - a connection to hematogeneous metastasis. J Biol Chem 287:2168-2178. Chien AJ*, Moore EC*, Lonsdorf AS, Kulikauskas RM, Rothberg BG, Berger AJ, Major MB, Hwang ST, Rimm DL, Moon RT. 2009. Activated Wnt/beta-catenin signaling in melanoma is associated with decreased proliferation in patient tumors and a murine melanoma model. Proc Natl Acad Sci USA 106:1193-1198. *equal contribution, alphabetical order Hedrick MN*, Lonsdorf AS*, Shirakawa AK, Lee CCR, Liao F, Singh SP, Zhang HH, Love PE, Hwang ST, Farber JM. 2009. CCR6 is required for IL-23-induced psoriasis-like inflammation in mice. J Clin Invest 119:2317-2329. *equal contribution, alphabetical order Huang V, Lonsdorf AS, Fang L, Kakinuma T, Lee VC, Cha E, Zhang H, Nagao K, Zaleska M, Olszewski WL, Hwang ST. 2008. Cutting edge: rapid accumulation of epidermal CCL27 in skin-draining lymph nodes following topical application of a contact sensitizer recruits CCR10-expressing T cells. J Immunol 180:6462-6466. Lonsdorf AS, Kuekrek H, Stern BV, Boehm BO, Lehmann PV, Tary-Lehmann M. 2003. Intratumor CpG injection induces protective antitumor T cell immunity. J Immunol 171:3941-3946 29 PACKAGE B2 4 Project Plan 4.1 Specific Aims 1. Characterization of CCR6/CCL20-dependent anti-tumor immune responses in primary malignant melanoma and skin-draining LNs in murine models of melanoma. 2. Identification of local immunmodulatory factors with functional relevance for CCR6/CCL20mediated anti-melanoma immune responses in primary tumors and skin-draining LNs. 3. Correlation of CCR6/CCL20 expression patterns in primary human melanoma and LN metastasis with respect to disease stage and local tumor progression. Project 12: Tumor-directed cytotoxicity of proinflammatory human dendritic cells and natural killer cells in malignant melanoma (MM) Principal Investigator: Prof. Dr. Knut Schäkel, Hautklinik, Universitätsklinikum Heidelberg; PD Dr. Adelheid Cerwenka; Boveri Research Group Innate Immunity, German Cancer Research Center London Project Partner: Prof. Frederic Geissmann, Center for Molecular and Cellular Biology of Inflammation, King’s College, London Short Summary: There is good evidence that dendritic cells (DC) and natural killer (NK) cells collectively mount a strong anti-tumor immune responses. However, we have a limited understanding of how these populations crosstalk with each other and how we can exploit the NK/DC interaction for the therapy of malignant melanoma. In addition, it is unclear which subtypes of DC are most relevant for inducing anti-tumor effector functions in the presence and absence of NK cells. A recent study from the Cerwenka lab demonstrated that the activation of NK cells with inflammatory cytokines such as IL12/15/18 greatly increases the anti-tumor activity of mouse and human NK cells. The subset of DC that produces the highest levels of IL-12 was identified CD11c+ slan (6-sulfo LacNAc+) DCs by the Schäkel lab. These slanDCs are equipped with an outstanding capacity to induce proinflammatory immune defence functions. slanDC were shown to exert a strong antibody-dependent (ADCC) and an antibody-independent tumor-directed cytotoxicity. They efficiently enhance the NKdirected anti-tumor cytotoxicity but the underlying mechanisms are poorly understood. We hypothesize that the NK/slanDC crosstalk is highly relevant for executing cytotoxic anti-tumor responses in the skin. In the proposed project, we plan to analyze the functional consequences and mechanisms of the NK/slanDC crosstalk in vitro and in a xenograft mouse model of human melanoma in vivo. In addition, we will analyse the presence of slanDC and NK cells within biopsies of melanoma patients after treatment with inflammatory agents such as application of the TLR-7 ligand Imiquimod. The results gained in this project could lead to novel therapeutic strategies for the treatment of MM based on the exploitation of the slanDC/NK crosstalk. PACKAGE B2 3 State of the Art 3.1 State of knowledge in the field The number, the type and the activation status of tumor-associated DCs and NK cells have been shown to be of direct prognostic value. Malignant melanoma cells often express low levels of MHC class I and high levels of activating NK cell ligands, and are therefore very efficiently killed by NK cells. In general, low numbers of NK cells are found in solid tumors but it has been shown that inflammatory agents such as the application of the TLR ligand CpG can facilitate NK cell infiltration into mouse solid tumors. DCs can mount a potent direct cytotoxic anti-tumor response. Cytotoxic DCs were shown to take the lead in inducing a cytotoxic anti-tumor response when MM were treated with the TLR7-ligand imiquimod. Furthermore, the antibody-dependent cell-mediated cellular cytotoxicity (ADCC) of DCs and NK cells contributes to the natural tumor surveillance and may significantly enhance tumor destruction in monoclonal antibody-based immunotherapies with e.g. trastuzumab. 3.2 Preliminary work by the participants Schäkel lab : We identified the population of slan (6-sulfo LacNAc+) DCs in humans (Schäkel et al., 2002). slanDCs are a population of proinflammatory DCs that stand out by their high level production of IL-12, IL-23, TNF-α and IL-1ß in response to TLR7 and TLR8 ligation (Hänsel et al. 2011 and 2012). slanDCs have a marked tumor-directed ADCC, and strongly enhance the cytotoxic capacity of NK cells (Schmitz et al., 2005). The role of proinflammatory or cytotoxic slanDCs in tumor tissue and their local interplay with NK cells has not been investigated so far. Cerwenka lab: Our previous study (Ni et al, JEM 2012) revealed that adoptive transfer of NK cells that were preactivated with IL12/15/18 resulted in greatly increased anti-tumor activity in mouse models of RMA-S lymphoma and B16 melanoma compared to NK cells pretreated conventionally 30 with IL15 or IL2. Importantly, human NK cells activated with IL12/15/18 also displayed sustained effector function and higher cell recoveries. To investigate the in vivo anti-tumor activity of human NK cells, we have established a mouse xenograft model in which luciferase-expressing human melanoma cells can be imaged in vivo after injection into NSG mice. In addition, our lab has a long-standing expertise in the investigation of activating receptors expressed by NK cells such as NKG2D and NKp30 and their ligands expressed by tumor cells (Textor et al, Cancer Res, 2011). 4.2 Experimental program 1. We will establish co-cultures of NK cells and DC with a focus on slanDCs. The crosstalk between these cells types in the absence and presence of TLR ligands will be assessed by cytotoxicity assays using melanoma cell lines that are well established in our laboratory as targets. Further experiments will assess the molecules involved in the NK/slanDC crosstalk (cell surface and soluble factors). 2. Next we will investigate the consequences of the NK/slanDC crosstalk in a xenograft model of MM. Co-cultures will be established in vitro and primed NK cells (or DC) will be adoptively transferred into melanoma-bearing mice. Tumor growth and functional parameters of the transferred cell populations will be monitored. In order to fully activate slanDCs, melanomas will be treated with Imiquimod. 3. We will conduct a comprehensive analysis to detect slanDCs and the NK cells by Tissue-FAX in tumor tissues. We will carefully determine the expression of parameters that provide information about the activation status (iNOS, TNF-α) and the maturational stage of slanDCs (CD83 versus CD206). The Heidelberg Tumor Registry will provide us with requisite tissue arrays. We will focus here on melanoma but will compare the obtained data with studies on SCC and basal cell carcinomas. Attention will be paid to spontaneously regressing tumors, and tumors regressing following TLR7-treatment. 4.3 Collaborations with other projects in the RTG For the evaluation of proinflammatory DC and NK cells in melanoma cells we will closly collaborate with project 13 (on myeloid cell subsets) and project 11 (on cellular recruitment by chemokines). 5 References 1. Ni J, Miller M, Stojanovic A, Garbi N, Cerwenka A. 2012. Sustained effector function of IL-12/15/18 preactivated NK cells against established tumors. J Exp Med 209:2351-65. 2. Textor S, Fiegler N, Arnold A, Porgador A, Hofmann TG, Cerwenka A. 2011. Human NK cells are alerted to induction of p53 in cancer cells by up-regulation of the NKG2D-ligands ULBP1 and ULBP2, Cancer Res 71:5998-6009. 3. Hansel A, Gunther C, Ingwersen J, Starke J, Schmitz M, Bachmann M, Meurer M, Rieber EP, Schäkel K. 2011. Human slan (6-sulfo LacNAc) dendritic cells are inflammatory dermal dendritic cells in psoriasis and drive strong T(h)17/T(h)1 T-cell responses. J. Allergy Clin Immunol 127:787-794. 4. Schäkel K, von Kietzell M, Hänsel A, Ebling A, Schulze L, Haase M, Semmler C, Sarfati M, Barclay AN, Randolph GJ, Meurer M, Rieber EP. 2006. Human 6-sulfo LacNAc-expressing dendritic cells are principal producers of early interleukin-12 and are controlled by erythrocytes. Immunity 24:767-777. 5. Schmitz M, Zhao S, Deuse Y, Schäkel K, Wehner R, Wohner H, Holig K, Wienforth F, Kiessling A, Bornhauser M, Temme A, Rieger MA, Weigle B, Bachmann M, Rieber EP. 2005. Tumoricidal potential of native blood dendritic cells: direct tumor cell killing and activation of NK cell-mediated cytotoxicity. J Immunol 174:4127-4134. 31 PACKAGE B2 4 Project Plan 4.1 Specific Aims 1. To determine the anti-melanoma activity induced by slanDCs/NK cell crosstalk in vitro. 2. To determine the melanoma-directed cytotoxicity induced by slanDCs/NK cell crosstalk in a xenograft mouse model in vivo. 3. To determine the presence and the activation state of proinflammatory slanDCs and NK cells in MM biopsies with and without treatment with the TLR7 ligand Imiquimod. Project 13: Regulation of tumor-associated macrophage and myeloid-derived suppressor cell activation and its neutralization in transgenic mouse melanoma model Principal Investigator: Dr. Astrid Schmieder, Dept. of Dermatology, University Medical Center Mannheim; Prof. Dr. Viktor Umansky, Skin Cancer Unit, German Cancer Research Center (DKFZ) and Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Ruprecht-Karl University of Heidelberg London Project Partner: Dr. Sandra Diebold, King's College London, Peter Gorer Department of Immunobiology, Division of Immunology, Infection and Inflammatory Diseases, Guy's Hospital, London Bridge, London Short Summary Melanoma immunotherapy is not satisfactory due to the accumulation of chronic inflammatory factors and immunosuppressive myeloid cells such as tumor-associated macrophages (TAM) and myeloid-derived suppressor cells (MDSC) in tumor lesions. The goal of the project is to better understand the molecular mechanisms underlying the inhibition of anti-tumor immune responses mediated by TAM and MDSC, and to design novel therapeutic strategies targeting these myeloid cells in melanoma. We will study the role of signaling molecules (p38 MAPK and S100A8/A9) and microRNA in the capacity of TAM and MDSC to inhibit anti-tumor reactivity of T cells. Results of the project will help to develop novel efficient human melanoma treatments neutralizing immunosuppression in the tumor microenvironment. PACKAGE B2 3 State of the Art 3.1 State of knowledge in the field Despite the intrinsic melanoma immunogenicity, immunotherapeutic trials were not satisfactory due to the formation of a complex immunosuppressive network mediated by chronic inflammation developing in the tumor microenvironment. Such microenvironment (represented by various cytokines, chemokines and growth factors) can induce a recruitment and expansion of suppressive immune cells such as TAM and MDSC to the tumor site. It has been shown that the expansion and activation of MDSC and TAM requires several signaling pathways (p38 MAPK as well as S100 calcium-binding protein A8 and A9). Small non-coding RNAs designated as microRNAs (miR) has been recently shown to significantly regulate the accumulation and function of TAM and MDSC. Thus, miR-494 can be induced in MDSC by tumor-derived TGF-β supporting immunosuppression and metastasis. On the other side, miR-511-3p was reported to modulate genetic program of TAM limiting their protumoral functions. Transgenic mice overexpressing human receptor tyrosine kinase Ret in melanocytes spontaneously develop skin melanoma with metastases in lymph nodes, lungs, liver, brain, and bone marrow and can be used for studying melanoma progression in vivo. 3.2 Previous/preliminary work by the participants To address the role of MDSC in melanoma progression in clinically relevant conditions, we used the ret transgenic mouse model, which mimics human melanoma with respect to clinical development ensuring natural tumor-stroma interactions. Analyzing Gr1+CD11b+ MDSC in melanoma lesions and lymphatic organs revealed a remarkable elevation of MDSC frequencies. Moreover, we found increasing concentrations of IL-1β, VEGF, IL-6 and GM-CSF in tumors during their progression. MDSC enrichment was accompanied by a decrease in TCR ζ-chain expression in tumor-infiltrating T cells. Therapy of melanoma-bearing mice with the phosphodiesterase-5 inhibitor sildenafil led to decreased numbers and immunosuppressive function of MDSC as reflected by the restoration of ζ-chain expression and significantly increased mouse survival. In addition, we identified in subcutaneous murine melanomas TAM that eco-express not only stabilin1 and LYVE-1 but also the novel surface marker MS4A8A, a molecule involved in differentiation processes. In vitro, these TAM were selectively induced via activation of the p38 MAPK and 32 glucocorticoid signaling pathways, indicating an important role of the p38 MAPK pathway also for the regulation of TAM activity. 4 Project Plan 4.1 Specific Aims Aim 1: Investigating signaling molecules and miR involved in the recruitment, expansion and activation of TAM and MDSC during melanoma progression Aim 2: Studying the neutralization of immunosuppression induced by TAM and MDSC using the modulation of their relevant signaling pathways and miR 4.3 Collaborations with other projects in the RTG For the evaluation of the recruitment and activation of TAM and MDSC in the melanoma microenvironment, we will closely collaborate with project 11 (CCR6/CCL20 interaction) and project 12 (proinflammatory dendritic cells). 5 References 1. Schmieder A, Schledzewski K, Michel J, Tuckermann JP, Tome L, Sticht C, Gkaniatsou C, Nicolay JP, Demory A, Faulhaber A, Kzhyshkowska J, Géraud C, Goerdt S. 2011. Synergistic activation by p38MAPK and glucocorticoid signaling mediates induction of M2-like tumor-associated macrophages expressing the novel CD20 homolog MS4A8A. Int J Cancer 129:122-132. 1. Schmieder A, Michel J, Schonhaar K, Goerdt S, Schledzewski K. 2012. Differentiation and gene expression profile of tumor-associated macrophages. Semin Cancer Biol 22:289-297. 2. Schmieder A, Schledzewski K, Michel J, Schönhaar K, Morias Y, Bosschaerts T, Van den Bossche J, Dorny P, Sauer A, Sticht C, Géraud C, Waibler Z, Beschin A, Goerdt S. 2012. The CD20 homolog Ms4a8a integrates pro- and anti-inflammatory signals in novel M2-like macrophages and is expressed in parasite infection. Eur J Immunol 42:2971-2982. 3. Meyer C, Sevko A, Ramacher M, Bazhin AV, Falk CS, Osen W, Borrello I, Kato M, Schadendorf D, Baniyash M, Umansky V. 2011. Chronic inflammation promotes myeloid-derived suppressor cell activation blocking antitumor immunity in transgenic mouse melanoma model. Proc Natl Acad Sci USA 108:17111-17116. 4. Zhao F, Falk C, Osen W, Kato M, Schadendorf D, Umansky V. 2009. Activation of p38 MAPK drives dendritic Cells to become tolerogenic in ret transgenic mice spontaneously developing melanoma. Clin Cancer Res 15:4382-4390. 33 PACKAGE B2 4.2 Experimental program 1. We will test the expression and activation (phosphorylation) of such signaling molecules as S100A8/A9 and p38 MAPK in F4/80highCD11b+Gr1- TAM and CD11b+Gr1+ MDSC in melanoma lesions and lymphoid organs using FACS. Differentially up- and down-regulated miR in TAM and MDSC will be studied by Affymetrix Microarray analysis and real-time quantitative PCR. Functional relevance of identified miR will be analyzed using lentiviral constructs. TAM and MDSC will be characterized by the expression of arginase-1 and inducible nitric oxide synthase. The activity of these myeloid cells will be determined by the inhibition of T cell proliferation upon the co-culture with TAM and/or MDSC. Inflammatory factors (VEGF, IL-1β, IL-6, TNF-α, TGF-β, etc.) will be detected in melanoma lesions by multiplex technology. Tumorinfiltrating T cells will be validated measuring the ζ-chain expression by FACS. 2. We will suppress signaling pathways involved in the expansion and activation of MDSC and TAM with specific inhibitors p38 MAPK (SB203580 and RO3201195). Activating miR will be blocked by the sponge preventing the interaction of this miRNA with its targets. TAM or MDSC isolated from melanoma lesions will be treated with these inhibitors followed by co-incubation with activated syngeneic T cells. T-cell proliferation and ζ-chain expression will be tested by FACS. 4 Qualification Program 4.1 Qualification Program Clinical researchers have to combine clinical expertise with a thorough understanding of the underlying molecular mechanisms that are altered in disease. Basic scientists with a future career in academic medicine (or affiliated research institutes) or in industry are more competitive if they have a sound clinical understanding of the diseases they work on. This is particularly true for cancer research and especially for the specialized field of dermato-oncology. The RTG will therefore establish a multi-disciplinary approach to teach cancer biology and dermato-oncology. To this end, the RTG will on the one hand build on the existing academic and teaching strengths of the Rhein-Neckar Region with several Graduate Schools of the Medical and Life Science Faculties of Heidelberg University and of the DKFZ, and on the other hand will offer additional teaching in a broad range of topics covering the diverse fields of clinical and experimental dermatooncology. With respect to existing Graduate Schools, Heidelberg University runs the Hartmut HoffmannBerling International Graduate School of Molecular and Cellular Biology (HBIGS) as part of the Excellence Initiative. The DKFZ runs the Helmholtz International Graduate School for Cancer Research (HIGS). Students in the RTG will be granted access to the two Graduate Schools, and the teaching offered by these schools will be integrated into the structured program of the RTG, which will focus on clinical and scientific aspects of dermatooncology. HBIGS will be responsible for teaching and training the PhD/MD students in general molecular and cellular biology and in “soft skills”, whereas HIGS will be responsible for the teaching program in cancer biology and general oncology. The added value the RTG will provide for the PhD/MD students will be the application of the general principles and approaches of cancer biology and oncology (the hallmarks of cancer) to the specific problems and questions of skin cancer biology and dermato-oncology (the hallmarks of skin cancer) as shown in Figure 1. Figure 1: Synergy between the RTG Hallmarks of Skin Cancer, the Hartmut Hoffmann-Berling International Graduate School of Molecular and Cellular Biology (HBIGS), and the Helmholtz International Graduate School for Cancer Research (HIGS). PROGRAM The teaching program for the RTG students will comprise regular seminars, courses, workshops, and scientific meetings that will be offered for PhD students in a structured sequence over a threeyear period. MD students will be enrolled into the RTG for 1 year and will participate in all teaching activities offered to the PhD students. Limitation of their term to one year will make careful selection of the available training opportunities necessary. This will be supported by mentoring and advice given by the responsible PIs and MD supervisors. The seminars will focus on general aspects of oncology (HIGS) and special aspects of dermatooncology (RTG), while the course/workshop program will offer methodological training in general techniques in cell and molecular biology (HBIGS, HIGS) and special techniques in dermatooncological research (RTG). The latter courses will include a mandatory 1-week laboratory teaching course per year. Soft skill training will be part of the program offered both by HBIGS and HIGS (Table1-3, see below). While the seminar series and the annual 1-week lab teaching courses will follow a given structured program, the courses and workshops may be freely chosen by the students according to their interests and educational needs. To allow the students of the RTG to fully exploit the teaching on offer, each RTG student will be allowed to attend all seminars, courses and workshops of both HBIGS and HIGS. Formally, each student will therefore become a member of either HBIGS or HIGS in addition to being a member of the RTG. The teaching program of the RTG reflects the basic idea that each area of the graduate school and their partners at the DKFZ (HIGS) and Heidelberg University (HBIGS) should equally contribute to the progress of the respective student/scientific group. Therefore, participation of all graduate students, associate students, and PIs in the teaching program is considered essential. Facilitating the communication with HBIGS, HIGS, and the RTG, several members of the RTG are 34 affiliated members of HBIGS (H.Augustin, Angel, Boutros, Leverkus, Goerdt, Sleeman, Schäkel, Utikal) and/or HIGS (H. Augustin, I. Augustin, Angel, Boutros, Utikal). 4.1.1 Seminars Table 1: Topics of the different Seminar Series (exemplified) STATE OF THE ART IN DERMATOONCOLOGY (RTG; PREFERENTIALLY 1ST YEAR) Several seminar series will be held to expose the students to all aspects of dermato-oncology. To start with, there will be a series of lectures on the principles of general and molecular oncology within the framework of HIGS (“Progress in Cancer Research Lectures”). Within a three-year term, this seminar series will introduce all departments of the DKFZ in Heidelberg and their research topics. Covering the different aspects of clinical and experimental dermato-oncology, the members of the RTG will give a structured seminar series over the course of the three-year program, starting with general aspects and problems of dermato-oncology in the first year. The lectures during the second and third year of the RTG series will be selected to preferentially deal with current topics in dermato-oncology and skin cancer. The seminar series will increasingly include the review of timely research papers pertinent to the projects of the RTG that will be performed by the students of the RTG, or may also represent current hot research topics (research lectures). The seminars of the 2nd and 3rd year will include presentations by international guest lecturers. In order to learn the conduct and the procedures of international collaboration in science, the students will be given the opportunity to invite, guide, and introduce the international speakers. When the seminar is given by a guest lecturer, the students will be given the possibility for discussion after the meeting (“meet the professor”, 0,05 ECP/h). A selection of topics of the seminar series is listed above (Table 1). 4.1.2 Laboratory Instruction Hands-on laboratory instruction will be provided by modular laboratory teaching courses (0,1 ECP/h). The annual 1-week laboratory course will be mandatory for all students (Table 2). 35 PROGRAM CURRENT TOPICS IN DERMATOONCOLOGY (RTG; PREFERENTIALLY 2ND AND 3RD YEAR) Immunotherapy of Skin Cancer Vascular Oncology Squamous Cell Carcinoma (Leverkus) (Umansky) Innate Immunity/Inflammation and Cell death pathways for skin Malignant melanoma (Utikal) Cancer tumorigenesis (Leverkus) Development of pigmentation, Cytoplasmic signaling Circuitry in Merkel cell carcinoma (Ludwig) pigment cell nevus and melanoma Cancer cells (Utikal) How to overcome cell death Cancer Genetics Cutaneous Lymphoma (Felcht) resistance in skin tumors (Geserick) General dermatopathology of skin Epithel-mesenchymal transition in Cancer Epigenetics tumors (Géraud) skin cancer (Sleeman) Oncogenic signaling in skin cancer Mitosis and Differentiation Mesenchymal skin tumors (Goerdt) (Boutros, Leverkus) Mechanisms of Apoptosis and cell Cell Migration and Tissue Formation Basal Cell Carcinoma (Leverkus) death resistance in lymphoma (Felcht) The concept of rational treatment of Vascular Oncology in skin cancer Mechanisms of Cell Death Resistance cancer (Utikal) (Augustin H, Schneider, Géraud) Tumor immunology in melanoma Oncogenic Signaling Rare cutaneous tumors (Ludwig) (Umansky, Lonsdorf, Schäkel, Enk) Innate Immunity/Inflammation and Animal models for studying melanoma Cancer Genome Biology Skin Cancer (Schmieder, Goerdt, (Sleeman) Schäkel) The lymphatic system in dermatoSpezialized dermatopathology of skin Stem Cells and Cancer Stem Cells oncology (Sleeman) tumors (Géraud) Animal models for the study of basal Stem cells in normal skin and skin Tumor Immunology cell carcinoma (Leverkus) cancer (Sprick) Animal models for studying squamous Mechanisms of metastasis formation Metastasis cell carcinoma (Angel) in skin tumors (Schneider) PROGRESS IN CANCER RESEARCH (HIGS) Table 2: Annual 1-week Laboratory Teaching Courses (mandatory) YEAR 1 YEAR 2 YEAR 3 BASIC LABORATORY TECHNIQUES IMMUNOLOGICAL METHODS ADVANCED LABORATORY TECHNIQUES DAY 1 DNA and Sequencing techniques Antibodies (generation, quality control and applications) Next generation sequencing DAY 2 PCR T-cells DAY 3 Basic biochemical Methods Dendritic cells/Macrophages DAY 4 Flow cytometry Chemotaxis, Adhesion and Transmigration High throughput RNAi techniques Transfection, transduction, and Reporter Gene Assays; Basic Proteomics CRISPR/Cas9 mediated mutagenesis DAY 5 Immunohistology/-fluorescence, LSM Animal / Genetic models in Cancer Immunology Generation of transgenic/knockout animals ORGANIZATION LEVERKUS MAHNKE I. AUGUSTIN/BOUTROS Numerous 2-day hands-on Laboratory Courses covering special topics will be available through HBIGS, HIGS, or the RTG (0,1 ECP/h; Table 3). In selecting lab courses, we will allow the graduate students a maximum of flexibility. The PIs will coach the students in choosing individual courses, but the individual selection process will also require that the students take responsibility to plan the educational program that will best fit their special interests and needs. This individually tailored modular system of block courses will allow the students to best take advantage of the multitude of offers provided by HBIGS, HIGS, and the RTG. While the strength of the HBIGS and/or HIGS courses are in the broad coverage of all technical aspects of basis research, the RTG will complement this laboratory instruction with topics relevant in skin cancer research. These RTG courses will be held in the laboratories of the PIs or co-PIs. Examples for such specialized dermatooncological lab techniques are tissue-specific culture of endothelial cells, organotypic cultures of skin cells, or methods for differential cell death detection (Table 3). In addition, the modular concept allows adaptation to the specific needs also of the MD students. Table 3: 2-day hands-on Laboratory Courses (Selection of Courses offered) GENERAL TOPICS IN CELL BIOLOGY AND MOLECULAR BIOLOGY (HBIGS) PROGRAM High-throughput screening for cancer target discovery TOPICS PERTINENT TO SKIN CANCER RESEARCH (RTG) How to isolate and work with human endothelial Cells (Felcht, Augustin) Isolation of primary lung microvascular endothelial cells from mice and rats (Géraud, Goerdt) Isolation of primary liver sinusoidal endothelial cells from mice and rats (Géraud, Goerdt) Confocal laser scanning microscopy Multicolor flow cytometry Advanced FACS sorting techniques Angiogenesis assays (Felcht, Augustin) 5 Multicolor FACS staining and sorting for stem cells/cancer-stem cells Lymphatic vessels (Wholemount staining, lymphatic ring assay) (Sleeman) How to simulate in vivo blood flow conditions by using microfluidic assays (Schneider) Multi Photon Microscopy Analysis of gene expression (RQ-PCR, in situ hybridization) Generation of IPS cells (Utikal) Rapid one-step mutagenesis of plasmid DNA Isolation and culture of primary melanocytes and melanoma cells (Utikal) High throughput screens Assays for tumor invasiveness (Sleeman, Angel) Orthotopic and Metastatic Mouse Models for Human Cancers (Sleeman) Genetic mouse models for malignant melanoma (Utikal, Umansky) Mouse models for squamous cell carcinoma (Angel) Analytical Ultracentrifugation Animal handling State of the art Imaging techniques Magnetic cell sorting Functional studies of human DCs (Schäkel) 36 4.1.3 Workshops The workshops serve to teach theoretical topics of general importance in pursuing research work, as well as key competences or “soft skills”. For most of the “soft skill” topics there will be courses offered within HBIGS and HIGS. Examples include learning strategies, scientific writing, presenting and rhetorical skills, interpersonal skills, entrepreneurship, and others. In addition there is a close cooperation with the Masters Program “Translational Medical Research” of the Medical Faculty Mannheim (coordinator Prof. J. Sleeman, also member of the Steering Committee of the RTG) that also offers a broad range of training in these “soft skills”. In order to allow the PhD students a maximum of flexibility for their experimental work, at least 3 workshops covering these “soft skills” topics should be attended during the 3-year schedule. 4.1.4 Student project development platforms and students’ conferences While preparing for the 2nd year British-German Workshop on Skin Cancer Biology (see 4.3), the students will write a structured progress report about their project. Within this progress report they will be encouraged to suggest how they envision the further strategic and experimental development of their projects. These project reports and continuation proposals will be discussed in detail at the Workshop with the PIs and with the London supervisors during the Project presentation sessions and at the individual TAC meetings. After the Workshop, the students will rewrite the proposals and make them available to the RTG community via the intranet for discussion. As an additional important active task, the students of the RTG will organize the 3rd year BritishGerman Workshop on Skin Cancer Biology as an international conference covering topics relevant to the “Hallmarks of Skin Cancer” serving also as a final status/farewell seminar. In this respect, they will be actively guided by the PIs/Steering Committee. 4.1.5 Transition from the first class of doctoral students to the next class Since the doctoral program is scheduled for 3 years, recruitment for the second generation of students will begin about 2.5 years after the start of the program. Before that, opportunity to join the programme will be given annually to associated doctoral students, for whom the same rules and rights will apply as for the regular students – except for funding by the RTG. Most parts of the program do not have to be taken in a specific temporal order. Those parts that have to be taken at the beginning or end of an individual schedule will be offered annually (legal, ethical matters). Each participating group will invite national and international expert scientists during the funding period. Emphasis will be put on individuals who have an outstanding reputation in the fields of skin carcinogenesis, skin tumor progression and metastasis, vascular biology, cell signalling, cancer target research and tumor immunology. Given the time constraints of top researchers, these visits will typically last for no longer than 3 days. The visiting program will also enable Professors/Lecturers from the St. John’s Institute of Dermatology and other groups in London to visit the PIs and graduates in Mannheim/Heidelberg, and consult about ongoing projects. 4.3 Additional qualification program: scientific collaboration with the London project partners of the RTG As science is an international endeavor, the RTG aims to introduce the students to international scientific collaboration. To this end, the RTG has set up the participation of an inter-institutional scientific faculty in the Metropolitan Area of London centered around the St. John´s Institute of Dermatology and King’s College. The faculty comprises high-ranking scientists of the St. John´s Institute of Dermatology, King’s College, the University of London and Cancer Research UK who are committed to the goals of the research program of the RTG, and who have outstanding expertise across the spectrum of relevant basic, translational, and clinical science. Each RTG project will be assigned a London project partner including Dr. Joy Burchell, Dr. Sandra Diebold, Prof. Anthony Dorling, Prof. Frederic Geissmann, Prof. Adrian Hayday, Prof. Frank Nestle, Prof. Fiona Watt, Prof. Sean Whittaker from King’s College; Prof. Fran Balkwill, Prof. Ian Mackenzie, Prof. Kairbaan Hodivala-Dilke from Queen Mary University of London; Prof. P. Meier from the Institute of Cancer Research; Prof. Henning Walczak, Prof. Buzz Baum from University College of London; and Dr. Ilaria Malanchi and Dr. Caroline Hill from London Research Institute, Cancer 37 PROGRAM 4.2 Guest scientist program Research UK. The input the London project partners will give to the scientific collaboration and the scientific carreer development of the students will be twofold: (1) Most importantly, the London project partners are committed to host the respective PhD/MD students for a variably long (6-week to 6-month), project-adapted lab visit to conduct part of the experiments for their thesis projects in London. This lab visit will heavily impact on the scientific development of the students offering them the opportunity to learn new techniques and to re-shape their projects due to the discussions with and the advice from the London project partner. (2) During the 3-year course of a PhD student’s life in the RTG, the RTG will organize a yearly “British-German Workshop on Skin Cancer Biology” in which the London project partners actively participate. In the first year, following recruitment of the students, the RTG will hold the 1st “BritishGerman Workshop on Skin Cancer Biology” in Mannheim to introduce the students to each other, to the Faculty of the RTG and to the London Faculty. The Workshop will include plenary lectures from selected London project partners, short project presentations by the students, the introductory course “How to design a scientific project”, and inaugural, parallel Thesis Advisory Committee (TAC) meetings. The 2nd “British-German Workshop on Skin Cancer Biology” will last 4 days, will be organized by the London partners in London and focus on the specific competence and scientific excellence of the London partners and their institutions. Table 4: Schedule of the 2nd-year “British-German Workshop on Skin Cancer Biology” in London 16 hrs= 1 ECP 1st day Key note lecture 9 10 11 2nd day Arrival 3rd day Lecture Cancer Stem Cells (F. Watts) Science structure and Funding opportunities in UK Coffee Break Selected project presentations Selected project presentations (incl. MD projects) 4th day Key note lecture γ δ T cells in tumor immunity (A. Hayday) 12 Key note lecture 13 Modern Methods in Translational Oncology (F. Nestle) Parallel TAC Meetings Lunch break 14 15 Key note lecture Selected project presentations (30 minutes per project) (10 min introduction PI, 10 min presentation of the graduate, 10 min plenary discussion) Site Visit Selected project presentations (London Partners) to the Francis Crick Institute Tumor Angiogenesis (K Hodivala-Dilke) PROGRAM 16 Site Visit 17 to the St. Johns Institute 18 Key note lecture Genetics of Skin Cancer (J. McGrath) Scientific Lectures (London Partners within the Departure Francis Crick Institute) All London project partners will attend this Workshop in London to allow for intense scientific and project-related interactions and discussions in the unique scientific environment of the London Faculty accompanied by the TAC meeting of the second year (see table 4). In the third year and as an additional important active task, the students of the RTG will organize the 3rd “British-German Workshop on Skin Cancer Biology” as an international 3-day scientific conference “Hallmarks of Skin Cancer” in Heidelberg which will also serve as a final status/farewell seminar. In this respect, they will be actively guided by the PIs/Steering Committee; the London project partners will also play an active role in this international scientific dermato-oncology conference. 4.4 Only Regarding International Research Training Groups: Research Stays at the Partner Institution Does not apply 38 5 Supervison and career development, equal opportunity / gender equality, organization, and quality management 5.1 Application and selection concept Recruitment of the best PhD and MD students is central to the success of the RTG. For the PhD studentships, the open positions will be advertised internationally via appropriate journals and scientific job platforms on the internet by the RTG in cooperation with HBIGS and HIGS. Each PI will also individually advertise his/her PhD position. The highly standardized admission procedures of HBIGS and HIGS will be extremely helpful for the selection of the best educated and most ambitious candidates, especially from foreign countries. Applications for each project either preselected by HBIGS / HIGS or on an individual basis will then be evaluated by the co-PIs as well as by representatives of the RTG (managing board) to ensure that standard high quality selection criteria are applied, including equal opportunity. For the MD recruitment process, the RTG will primarily rely on the local MD/PhD programs to identify the top MD students for a 1yr thesis project, including the scientific part of the Reformed Medical Study Program of the Medical Faculty Mannheim, MARECUM, i.e. the Junior Scientific Master Class Program and the Translational Medical Research Master Program (TMR). MD students are increasingly difficult to recruit for ambitious doctoral programs, and we have therefore only included 8 MD positions per year. Supervision, mentoring and RTG contract. A structured and individualized supervision concept is the basis for the success of the RTG. All rights and liabilities of the RTG and the student will be fixed in a contract signed by the university and the student at entry. The supervision concept will include the established elements of daily/weekly PI supervision with the active contribution of graduate students in the weekly group seminars to develop presentation skills and scientific discussion abilities. The supervisors will meet at least once monthly with each student for intense exchange and project-specific discussion. Each student will choose a co-supervisor from the RTG with whom the project progress will be discussed at least annually. The Speaker of the RTG will keep a permanent record of the credits earned by each student, and will share responsibility for the progress of the student with the supervisors. Additional external mentoring will be provided for each graduate student by assigning them an external co-supervisor within the framework of the collaboration with the inter-institutional London RTG Faculty. This mentoring program will include individual counseling during the British-German Workshops on Skin Cancer Biology and the lab visit to the external mentor’s laboratory. In case of the need for more intense or specific counseling, individual meetings with the London co-supervisor will be arranged. This multiple level mentoring program will facilitate continuous and focused supervision of the graduate student, and will allow timely intervention to guarantee the graduate student’s best scientific and educational development. Students’ Performance. To continuously evaluate the progress of the student, the RTG will implement a credit point system. The graduates will be expected to collect a defined number of credit points (60 ECP) per year. Should a graduate student not obtain the required number of points, the PI and the Speaker of the RTG will be responsible to interview the student. It is expected that the graduate students devote approximately 10 - 15% of their working time to the structured educational program of the RTG. Altogether the graduate students should earn at least 180 ECP during their 3-year thesis work. Of these, 50 ECP will be granted annually for the continuous experimental work on the thesis project, while 10 ECP annually must be collected by participation in the educational program. Depending on the degree of passive or active participation, 0.05 or 0.1 credit points will be accredited per hour. Similar to the ECP system of the RTG, HBIGS and HIGS use ECP to assess the achievements of the students. ECP earned in courses held by HBIGS or HIGS will be fully accepted by the RTG. MD students admitted to the RTG will have to earn 60 ECP during their one-year thesis work. Scientific Independence. Scientific independence is one of the major goals of the supervising concept of the RTG. This will be specifically supported by the modular teaching system, and complemented with an obligatory introductory course ("how to design a scientific project"), presentation and discussion of the project outline during the 2nd-year British-German Workshop on Skin Cancer Biology, and regular progress reports. The students will be encouraged to actively engage in the organization of a meeting covering the focus of the RTG. Furthermore, relevant 39 PROGRAM 5.2 Supervising Concept and Career Development national and international meetings will be announced during the teaching events and via the intranet, and the graduate students will be encouraged and financially supported to participate in such meetings and to present their data. Start-up Grants. Within the framework of the RTG, the most brilliant students will be actively encouraged to remain within the scientific field of dermato-oncology. The RTG will therefore offer doctoral students the opportunity of applying for start-up grants when they approach the end of their PhD work. A transparent application procedure will be implemented to allow these students to fund their own start-up projects through RTG funds (see 7.8). The previous PI and the speakers/Steering Committee will closely counsel the applicants. The primary goal of the start-up grant is to generate sufficient peer-reviewed scientific evidence to support the initiation of an independent research career. A primary evaluation criterion of start-up grant applications will thus be whether the proposed project will generate sufficient results to successfully support a subsequent application for grant funding from the DFG. Start-up grants will either fund local independent research by the start-up applicant, or will pave the way to other international laboratories in which the start-up applicant will further deepen his/her knowledge in the selected scientific field of dermato-oncology. 5.3 Equal opportunity / Gender Equality in Science The RTG „Hallmarks of Skin Cancer“ will join and strengthen ongoing activities in Heidelberg University and the German Cancer Research Center (DKFZ) to balance the relation between male and female researchers and to support the compatibility of work and family responsibilities. It will especially take measures to systematically enhance career development and qualification of young female scientists. The aims and the status quo regarding the numbers of femal and male participating MD/PhD student and researchers are found in tables A and B. A. MD/PhD students % Aim MD/PhD students m f (40-60 %) (40-60 %) PROGRAM Measures already taken by the University of Heidelberg. The PIs, postdoctoral fellows and the B. Participating Researchers students of the RTG „Hallmarks of Skin Cancer“ will be Number % supported by the ’Gleichstellungsbüro’ (Equal Status Quo Status Quo Opportunities Office) of the University, by the Equal Opportunities Offices of the Medical Faculties and of the m f m f DKFZ. Here many measures are already being taken to realize gender equality, to enhance the careers of Junior PIs 3 3 50 50 women especially in their initial stages, and to support female and male scientists with children. The University Senior PIs 12 1 92.3 7.7 has implemented a gender action plan that specifies targets and management by objectives in order to 15 4 79 21 steadily improve gender equality in all departments. We Total refer to http://www.uniheidelberg.de/einrichtungen/gleichstellung.html for a comprehensive documentation. According to the recent evaluation by the DFG, Heidelberg University has reached Step 4 regarding the implementation of the “Forschungsorientierte Gleichstellungsstandards der DFG”.In particular the following measures have already been realized within the above institutions and all members of the RTG agree to participate. The Olympia Morata Program of the University of Heidelberg supports “habilitations” of female scientists by financing 0.5 VK of their own position (TVL E13) reducing their routine workload. The program is open for participation by the female members of the RTG „Hallmarks of Skin Cancer“, and it will enable them to continue and advance a career in science. A specific mentoring and training program for female researchers within the life sciences is organized by the equal opportunities commissioner in cooperation with the Faculties of Medicine and the DKFZ, which targets the transition phase from the doctoral to the postdoctoral period, and aims to educate and prepare women for leadership in a career in science. Furthermore, various training events and meetings for young scientists such as Wi MEET are organized to support the networking of young female scientists in order to integrate them into the scientific community. 40 Measures to support the career of scientists with children. By offering a total of 407 places for children (in the age range from 2 months to 6 years) Heidelberg University wants to attract more female scientists to long-term academic careers. In cooperation with the University, the RTG „Hallmarks of Skin Cancer“ will offer custom-made services to support young scientists who have a family. This includes both full-time child care (Kinderhaus) and a ‘just in time’ or ‘back-up’ care initiative (KidsClub). Currently, the RTG has reserved places especially for use by its members. The RTG „Hallmarks of Skin Cancer“ is thus well prepared to provide child care services to its doctoral researchers, and its members can also participate in the University’s Concierge Service, which helps to reconcile academic work and domestic duties.. Measures already taken by the German Cancer Research Center and HIGS. Since 2005, the DKFZ has implemented an “Audit Work und Family” (Audit Beruf und Familie) procedure, a strategic management tool to help facilitate a better work-life balance for employees. Numerous opportunities are offered to all members of the DKFZ to promote compatibility between work and family life, and equal opportunities. These include child-care places, a mother-child room, mentoring programs at several levels of the career and “dual-careers” support. The gender ratio of HIGS is well balanced (57% female). However, despite the high representation of females among the students, the number of women in leading positions at the DKFZ is much lower. At HIGS, this imbalance is openly discussed and addressed during the PhD training. Support for women in science is provided by the DKFZ Executive Women’s Initiative, which focuses specifically on requirements of female scientists as they seek to obtain leadership positions in science. Several HIGS-specific measures exclusively for PhD students exist to ensure work-life balance and equal opportunities. HIGS currently offers up to six subsidized child-care places to PhD student parents at the Glückskinder day-care center close to the DKFZ. Furthermore, the PhD Careers Service offers a large range of courses and seminars to PhD students, encompassing themes relating to work-life balance planning and promotion of women in science. Measures made for and to be realized within RTG „Hallmarks of Skin Cancer“ All members of the RTG will be encouraged to participate in the above-mentioned programs for gender equality and family support offered by Heidelberg University and the German Cancer Research Center, and will receive financial support to do so. In addition, the following measures will be realized as part of the RTG program. • • • Recruitment and Motivation of Female Candidates. During recruitment of candidates for the PhD/MD positions the RTG will clearly announce that female applications are highly welcome and that the RTG especially supports the career development of female scientists. All members of the consortium will pay particular attention to recruiting talented female undergraduates and MD students to the RTG who attend their lectures. Networking. In close cooperation with the RTG, the University’s Graduate Academy and the equal opportunity commissioners of the University and the DKFZ will support the organisation of regularly held workshops to bring the female students of the RTG into close contact with a network of experienced female postdoctoral researchers in and beyond the field. International Orientation. The RTG will provide dedicated financial support for visiting female researchers to foster exchange of scientific ideas and to facilitate international networking. These meetings will provide the opportunity for female students to get directly in touch with leading international female scientists. This will have a strongly inspiring and mentoring effect on our female doctoral and postdoctoral researchers, in line with the strategic targets set by the RTG and the University. The RTG will furthermore cover travel expenses for female PhD / MD 41 PROGRAM The Medical Faculty Mannheim (Department of Gender Mainstreaming) has offered professional consulting for women, parents, students, colleagues in difficult life situations or with individual problems since 2008. Part-time work and home office work are possible. Day-care for children aged 0 – 3 started in 2009 (20-22 children) and day-care for children aged 2 – 6 (15-20 children) in 2010. The day-care named MEDI-KIDS is available daily for 12 hours and only closes for 10 days a year. During holidays, children aged 7 – 12 are invited to join a summer sport camp (DELTA-KIDS). 2012 a student baby-sitter-service was successfully implemented for members of the faculty. The RTG requests financial support for gender measures which include various procedures such as technical support, mentoring activities and complementary child care. • students who wish to present their research results and prepare for a subsequent research stay in a laboratory led by a female scientist who will serve as a role model. Postdoctoral Support. In order to achieve sustainable effects on career planning and gender balance, the RTG will provide additional support for the first postdoctoral period of alumni after their PhD thesis work, with a special focus on gender equality. This support will be awarded on the basis of a convincing and tight research plan that has been approved by the scientific committee and supervisors under the auspices of the start-up grants (see 5.2). Funding by the gender balancing program of the RTG „Hallmarks of Skin Cancer“ The RTG will provide financial support for the following gender equality measures: • Child care and measures to enhance the compatibility of study, research and family. • Support by technical assistance in case of long absence (illness, pregnancy). • Invitation of outstanding female researchers for national and international exchange and networking with the young talented female participants in the RTG program. • Seminars preparing female researchers within the RTG program to continue their career far beyond their PhD and helping them to plan the elements that make up a career in academia, and to get education for the needed specific skills. 5.4 Organization To manage the complex task of inaugurating and administering the novel RTG, a three-level infrastructure will be established comprising the Managing Board, the PI Convent, and the Students’ Assembly and Students’ Representatives, and other forms of student participation. PROGRAM The Managing Board of the RTG comprises the speakers, the secretary, three elected representatives of the PIs, and three elected representatives of the graduates. The board will meet every three months on a regular basis and on demand. The board of the RTG will organize the yearly PI convent and a yearly full assembly of all graduates to discuss administrative matters. The speaker of the RTG will be responsible for preparing strategic decisions and communication with the partners in London, as well as with the PIs and the students’ representatives regarding general matters. The speaker will also prepare the evaluation of the applications for the doctoral positions within the RTG to be decided by the Managing Board and the respective PIs. The vice-speaker will serve as a general coordinator. He will be responsible for the day-to-day organization of the RTG. The secretary of the RTG will serve as a personal assistant to the speaker and the vice-speaker. She/he will also be actively involved in the organization of the study program, including lectures and courses and the interaction with the officials in London in organizing the British-German Workshops on Skin Cancer Biology. He/she will be involved in the inauguration of a structured procedure for enrolling and registering new graduates, and will take responsibility for the following issues: studentship contracts, insurance, payment logistics, and short-term housing (for visiting students and teachers). The Speakers will also evaluate the applications for the rotational positions and will make a granting decision. Together with the speakers, the secretary will manage the accountancy of the 13 projects, the budgeting of the teaching program and the settlement of accounts with the DFG. Together with the vice-speaker and the PIs, the secretary will be responsible for coordination and organization of lectures and symposia (correspondence regarding invitations, transfer, housing logistics), establishing and updating the homepage of the RTG (together with the IT-Manager of the Faculty), design of invitations, brochures, information, statistical work regarding quality management and students’ affairs, and protocols of meetings. The PI Convent. The PIs of the RTG projects are full members of the PI Convent, while the associated PIs are members without the right to vote. Members of the London Faculty are not formal members of the PI Convent. The PI Convent will elect the three representatives of the PIs to serve on the Managing Board. The PI Convent will be responsible for deciding general matters of strategic importance for the RTG including recruitment of new PIs or Projects, and for the evaluation of the outcome of the RTG and the students’ performance. The PIs will be invited to convene on a regular yearly basis and on demand by the speaker, and will be informed by the speaker about the progress of the RTG, current requirements, and future meetings. The results of these meetings will be regularly communicated to all members of the RTG by the secretary via the webpage of the RTG. 42 Students’ Assembly, Students’ Representatives and other forms of participation. After the first 4-8 weeks on campus, the new class of students will convene for a meeting to discuss organizational matters of the RTG, and the details of the study program. The election of 3 students’ representatives who will be members of the Managing Board, take part in the PI Convent and report problems and suggestions concerning the whole class to the speakers of the RTG will also take place at this meeting. The students’ assembly will convene at least once a year and on demand. The students’ representatives will be (re-)elected every year. Internal Communication and Public Relations. Effective dissemination of information will be achieved via the intranet/internet. We will establish a RTG website that contains all information about ongoing teaching and research activities (e.g. by virtual posters, alerts for recent publications) and a newsboard with daily updates. This activity will be part of the responsibilities of the Secretary of the RTG supported by the PR department of the Medical Faculty in Mannheim (Frau Dr. Wellnitz). In addition, the website will contain a secured internal discussion forum accessible to RTG members only. In addition, the webpage will contain material and links for informing the public about skin cancer in general and about the activities of the RTG. 5.5 Additional aspects of quality management The dynamics of research demand a highly adaptive frame in which young researchers can develop and mature. Thus, specific measures will be implemented to meet the steadily changing challenges and needs in the research and qualification program. 1. At the regular yearly British-German Workshops on Skin Cancer Biology, there will be intense and structured discussions about the progress of the research projects and their future directions with all members of the RTG and the foreign mentors in London which will give the PhD/MD students and their PIs as well as the Steering Committee (see Section 2) a continuing feedback to improve and re-direct the projects. For the selection of MDs, the PI will be obliged to submit a MD project outline to the steering committee for approval and criticism. Additional candidate PhD/MD projects for inclusion in the RTG will be reviewed by the Steering Committee and an external reviewer from the London RTG faculty. Candidate projects should fulfil the criteria of scientific focus (dermato-oncology), excellence (publications, track record, extramural funding), qualification (successful thesis supervision), and teaching activities (workshops, seminars, clinical expertise). After approval, these projects will be considered as associated projects and will contribute to the further development of the research program of the RTG. 43 PROGRAM Other forms of participation. Following recruitment of the students, the RTG will organize the 1st year British-German Workshop on Skin Cancer Biology, a mandatory inaugural two-day meeting to introduce the students to the RTG and to each other as well as to the British PIs. This 1st year British-German Workshop on Skin Cancer Biology will serve to present and discuss the scientific projects and their status at the beginning of the RTG with the members of the RTG as well as with the British Co-PIs; it will furthermore include general instructions, and the introductory course “How to design a scientific project”. PIs, associate PIs, students, and the members of the Managing Board will meet on a regular weekly basis to discuss ongoing issues of the RTG and of the students’ career development in the form of an open, informal “jour fixe” (Wednesday evenings after the seminar series). Such a weekly meeting of the students together with some of the supervisors will allow for closer interactions and the initiation of fruitful collaborations within the frame of the RTG. When the first students are on the brink to graduate, the 3rd year British-German Workshop on Skin Cancer Biology will be organized as an international students’ conference/final status/farewell seminar by the PhD/MD students. The RTG will register all PhD/MD students into either HBIGS or HIGS. HBIGS and HIGS offer scientific platforms for multiple additional courses that cover a broad spectrum of areas within cellular and molecular biology of interest to the PhD students of the RTG. Similar to the ECP system that will be implemented within the RTG, HBIGS uses ECP to assess the participation of the students within the teaching program. Admission of the PhD student to HBIGS will give the PhD students a maximum of flexibility while profiting from the broad spectrum of educational programs in the Rhein-Neckar region. 2. In order to continuously improve the qualification program, feedback will be provided by the graduates at the regular yearly British-German Workshops on Skin Cancer Biology as well as during the regular jour fixe, the meetings of the Managing Board and the Steering Committee as well as in the Students’ Assembly. On this basis, the Managing Board will develop measures to improve the qualification program and discuss these measures with the PI convent and the students’ representatives before implementation. 3. Structured data documentation will be implemented from the start of the RTG, and will cover the following areas: a. application and selection procedures; b. scientific meeting contributions (meeting abstracts); c. CV-relevant graduate achievements (publications, grants, awards), d. alumni program (documentation of postdoctoral positions). 4. Success will be measured using the following parameters: adherence to the 3-year qualification period for PhD students (TTT – time to thesis), numbers of finished PhD / MD theses, publications in peer-reviewed journals, approved project grants by national/international foundations, travel grants to congresses, poster awards at conferences, congress participation by invitation. 6 Scientific Environment PROGRAM Cancer is without doubt the major clinical and research focus of both medical faculties of Heidelberg University. Oncology as a research focus of the University is structurally strengthened by the close interaction with the German Cancer Research Center in Heidelberg (DKFZ) including the DKFZ-ZMBH Alliance, by the National Center for Tumor Diseases (NCT) and by the German Consortium for Translational Cancer Research (DKTK). Heidelberg University together with the DKFZ is also a unique place to realize the research aims of the RTG as here is the highest concentration of research and clinical departments devoted to dermatology in general and specifically skin cancer in Germany. The Depts. of Dermatology in Heidelberg and Mannheim with Sections for Experimental and Molecular Dermatology, both being certified Skin Cancer Centers, the Clinical Cooperation Unit Dermato-Oncology of the DKFZ in Mannheim, and the Dept. of Signal Transduction and Growth Control of the DKFZ are all primarily devoted to research in dermatooncology. Basic research departments of both faculties and the DKFZ with a major research interest in skin cancer biology, such as the Depts. for Vascular Oncology / Tumor Angiogenesis and of Signaling and Functional Genomics, further strengthen this Heidelberg-Mannheim Skin Cancer Alliance. Despite its major focus on cancer, Heidelberg University does not run a Collaborative Research Center (SFB), an excellence cluster, another research training group (RTG) or a graduate school directly in the field of oncology with the exception of the TRR77 “Liver Cancer” which however is in its running out phase. The SFBs of the University in the field of the life sciences are rather organized in a mechanism-oriented manner such as the TRR23 “Vascular Differentiation and Remodeling”, the SFB 873 “Maintenance and Differentiation of Stem Cells”, and the SFB 938 “Enviroment-dependent Control of Immunity”. With respect to graduate schools, Heidelberg University runs HBIGS as part of the Excellence Initiative, and the IRTG 1874/1 “Diabetic Microvascular Complications / DIAMICOM” in Mannheim, while the DKFZ runs HIGS to serve graduate students with a DKFZ-affiliated PI. In order to assure the excellence of RTG students, admission via HBIGS to either HBIGS or HIGS will be a requirement. This will also allow the PhD students to obtain the “Dr. rer. nat.” from the Heidelberg Faculty of the Sciences. Several members of the proposed RTG actively participate in each of the SFBs / graduates schools mentioned above. HBIGS/HIGS offer an excellent administrative platform and several of the PIs of the RTG are members of HBIGS and/or HIGS. This should help to recruit the best candidates for the RTG. Of course, when granted, the RTG would also apply to be officially recognized as a structured PhD program by the Heidelberg Faculty of Biosciences. The RTG will ensure that graduate students can take courses in the IRTG “DIAMICOM” and the integrated research training group of TRR23. This will give them a maximum of learning possibilities together with the flexibility required for efficient and goal-oriented research practice. The added value of the RTG is not only a better education of PhD and MD students in the field of skin cancer that will ensure to attract young researchers into this field, but it will also bring skin cancer research departments in Heidelberg-Mannheim into a closer collaboration than hitherto established. Such a truly developed Heidelberg-Mannheim Skin Cancer Alliance may in the future be the basis for a skin cancer-oriented collaborative research center application to the DFG. 44 6.1 Demarcation from existing SFBs The RTG does not overlap with any SFB at Heidelberg University. In contrast to the RTG “Hallmarks of Skin Cancer”, SFB 850 “Control of Cell Motility in Morphogenesis, Cancer Invasion and Metastasis” at the University of Freiburg concentrates on general mechanisms of physiological as well as tumor cell motility; skin cancer plays but a minor role in this consortium. In contrast to the RTG, SFB 773 “Understanding and Overcoming Therapy Resistance in Solid Tumors” at the University of Tübingen (run out since June 30, 2013) has focused on analyzing and evading the mechanisms of secondary tumor resistance to established therapies such as chemo- and radiotherapy; skin cancer played but a minor role in this consortium. The Melanoma Research Network of the Deutsche Krebshilfe is a nation-wide research consortium comprising projects that address most aspects of melanoma initiation and progression. In contrast to the RTG, cutaneous squamous cell carcinoma is not included, and the members of the Heidelberg/Mannheim Skin Cancer Alliance at present do not hold funded projects in the Melanoma Research Network. Furthermore, the RTG offers a structured supervision and qualification program for PhD/MD students while the Melanoma Research Network as a nationwide research consortium does not have such a focus. Therefore, there is no direct overlap of the RTG with the Melanoma Research Network, but the two consortia complement each other and will together considerably strengthen the overall impact of skin cancer research in Germany. 6.2 Demarcation from preexisting graduate colleges Does not apply 7 Modules / Requested funding 7.1 Module Research Training Group 7.1.1 Funding for Staff 7.1.1.1 Funding for PhD students 12 PHD POSITIONS EUR 458.640 per year The scientific environment of the proposed RTG is highly competitive with two medical faculties in Heidelberg and Mannheim, the Faculty for Bioscience in Heidelberg, the DKFZ in Heidelberg, the EMBL, the Max-Planck-Institute for Medical Research as well as a considerable number of biomedical and pharmaceutical companies in the (Bio) region. This includes a considerable number of collaborative research centers, (integrated) research training groups and institutional graduate colleges/schools. The cost of financial support for a PhD student differs considerably, ranging from € 1.100 (Medical Faculty Mannheim) for a fellowship, and from 0.5% TVL E13 to 0.65% TVL E13 (SFB938, TRR23) for PhD positions. Most of the PhD students in the Heidelberg/Mannheim area are offered TVL positions. To ensure maximal attractiveness of the new RTG positions and high profile recruitment of the best students to do their research work and biomedical training in the new RTG, we are applying for the most attractive funding option, i.e. 0.65% TVL E13 positions. This will be an essential incentive to out-compete the high number of other attractive offers from well-renowned laboratories in the Heidelberg-Mannheim area. As the success of the RTG will highly depend on the quality of the recruited PhD students, it is crucial for the RTG to make every effort to attract PhD students from among the top 10% of their classes. 7.1.1.2 Funding for MD students The funding for the 8 MD students of the RTG à € 670,-- for 4.5 years each will be provided by the Medical Faculties Heidelberg and Mannheim and by the DKFZ. Five MD stipends will be funded by the Medical Faculty Mannheim, two MD stipends will be funded by the Medical Faculty Heidelberg, and one MD stipend will be funded by the DKFZ. 7.1.1.3 Funding for postdocs Does not apply 45 PROGRAM 12 x 0.65 VK TVL E13 for 4.5 yrs 1 PhD position funded by the DKFZ for 4.5 yrs 7.1.1.4 Qualification fellowships Does not apply 7.1.1.5 Funding for research students and pupils Does not apply 7.1.2 Consumables and Further Funding 7.1.2.1 Consumables and Small Equipment CONSUMABLES EUR € 16.500,- per PhD student per year for 4.5 yrs 214.500 per year Small equipment Does not apply 7.1.2.2 Travel costs Travel costs for PhD students FUNDING FOR EUR PROGRAM 2 British-German Workshops on Skin Cancer Biology in London (Flight to London (one way) 13 PhD students, 8 MD students; 2 times € 300,- per PhD/MD student Bus trip London – Mannheim (100,- € person) Accommodation (€160/d) and per diem (€47/d) London for 4 days, per PhD/MD student Individual visits to London for scientific collaboration within the co-PIs’ labs in the 2nd year of the course (13 PhD students, 1.5 student generations) Flight to London € 570,-- per student Duration of the lab visit in London from 6 weeks to 6 months, mean 4 months Auslandszuschlag (€ 779 per month) and Kaufkraftausgleich (€ 238 per month) in analogy to the calculations for stipends; € 1.017 per PhD student per month Individual visits to London for scientific collaboration within the co-PIs’ labs (8 MD students, 4.5 student generations, of these estimated 25%) Flight to London € 570,-- per student Duration of the lab visit in London from 6 weeks to 3 months, mean 2 months Auslandszuschlag (€ 779 per month) and Kaufkraftausgleich (€ 87 per month); € 868 per MD student per month Travel to scientific meetings One meeting per student à € 500,-- (13 PhD, 8 MD) per year for 4.5 years 12.600 4.200 34.776 11.115 79.326 5.130 15.588 10.500 per year Travel costs for principal investigators FUNDING FOR EUR 2 British-German Workshops on Skin Cancer Biology, London Flight to London and travel tickets € 570,-- per PI (13 PI, 2 times) Accommodation (€160/d) and per diem (€47/d) in London for 4 days, per PI (13 PI, 2 times) 46 14.820 21.528 7.1.2.3 Funding for Visiting Researchers FUNDING FOR EUR Guest lecturers (for 1-day seminars; € 500,-- travel costs, € 200,-- accommodation, € 300,-- salary) 10 times per year for 4.5 years 10.000 7.1.2.4 Animal Costs Animal costs will be supported by institutional core funding 7.1.2.5 Further funding Does not apply 7.1.2.6 Publication costs EUR 20.000 7.2 Module Substitute Does not apply 7.3 Module Coordination COORDINATION OF THE RTG EUR 0.5 VK TVL E8 for 4.5 yrs 21.900 per year Foreign Language Secretary (for coordination, students affairs, cooperation with HBIGS, organization of lectures, seminars, workshops, summer schools, contacts to London partners, support for gender equality issues, finances, accounts, homepage of the RTG, effectivity control, see this application 5.4.a) 7.4 Module Rotational Positions 2 ROTATIONAL POSITIONS EUR 168.000 per year The proposed RTG heavily relies upon a successful interaction of physician scientists and basic researchers as mentors for the PhD and MD students. Of the 19 PIs and co-PIs, 11 PIs within 9 projects are physician scientists with a considerable clinical workload. With two rotational positions, 8-9 physician scientists within the 9 projects led by clinical PIs could devote one year totally to research and supervision of doctoral students in the RTG. Interested physician scientists involved in the research projects of the PhD or MD students in the RTG are invited to submit an application to the Speakers of the RTG annually for a one-year rotational full time/half time position with a project proposal, full CV, publication list, and list of extramural peer-reviewed funding. The Speakers of the RTG will evaluate the applications and will make a decision based upon criteria of scientific excellence. As suggested by the DFG, the Medical Faculty Mannheim will finance one of the rotational positions from the Grundausstattung provided the other position is financed by the DFG. 7.5 Module Mercator Fellows Does not apply 7.6 Module Project-Specific Workshops BRITISH GERMAN WORKSHOPS ON SKIN CANCER BIOLOGY 2 1st year Workshops in Mannheim à € 10.000 each 1 3rd year International Workshop in Heidelberg 7.7 Module Public Relations Does not apply 47 EUR 20.000 30.000 PROGRAM 2 VK TVÄ TdL Ä1/2 for 4.5 years 7.8 Module Start-up Grants START-UP GRANTS EUR 2018-2019 100.000 In order to allow doctoral students from the RTG to continue their research projects after finalization of their PhD project and to develop their own research project thereafter, we apply for € 100.000,00 of “Start-up Grants” in the years 2018 to 2019. The application and selection procedures are detailed in Section 5.2 “Start-up Grants”. 7.9 Module Equal Opportunity / Gender Equality EUR 15.000 per year Support is requested to specifically aid female PIs and graduates or PIs and graduates with children if required. Examples are support for experimental work, mentoring activities, supplementary child care. Table 1: Does not apply Table 2: HOURS AS STAFF PERCENTAGE OF FULL TIME NUMBER DURATION (FROM – UNTIL) TVL E13 65% 12 01/04/2015-30/09/2019 - - - TVÄ TdL Ä1/2 100% 2* 01/04/2015-30/09/2019 Module Research Training Group PhD Student Postdoctoral Researcher Module Rotational Positions PROGRAM *As suggested by the DFG, the Medical Faculty Mannheim will finance one of the rotational positions from the Grundausstattung provided the other position is financed by the DFG. Table 3: 2015 FROM APRIL 2016 2017 2018 2019 TILL SEPT. - - - - - SUM Module Research Training Group Support Staff (Student Assistents) - Equipment up to € 10.000, Software and Consumables 160.875 214.500 214.500 214.500 160.875 965.250 Travel 11.328 89.213 45.251 15.104 85.437 246.333 Visiting Researchers 7.500 10.000 10.000 10.000 7.500 45.000 Experimental Animals - - - - - - Other - - - - - - Publications - - 5.000 10.000 5.000 20.000 48 Module Substitute - - - - - - 16.425 21.900 21.900 21.900 16.425 98.550 - - - - - - 10.000 - 30.000 10.000 - 50.000 Module Public Relations - - - - - - Module Start-up Grants - - - 50.000 50.000 100.0010 Module Gender Equality Measures in Research Networks 11.250 15.000 15.000 15.000 11.250 67.500 SUM 217.378 350.613 341.651 346.504 336.487 1.592.633 Module Coordination Module Mercator Fellows Module Project-specific Workshops (All figures in Euro) 8 Only regarding International Research Training Groups: Complementary Funding by the partner institution Does not apply 9 Declarations 9.1 Relations to other SFBs Does not apply 9.2 Collaboration with other cooperation partners 9.3 Cooperation with corporate partners Does not apply 9.4 Admission of qualification students Does not apply 9.5 Submissions of the proposal to other funding organisations Does not apply 9.6 Only regarding International Research Training Groups: Letter of Intent of the partner institution Does not apply 49 PROGRAM Letters of Intent from the medical faculties of Heidelberg University regarding funding, and of the London Coordinator, Prof. A. Hayday (King’s College and Cancer Research UK, London) regarding scientific collaboration, are to be found in the supplementary part of the application. 10 Obligations In submitting this proposal for an RTG to the DFG, Heidelberg University and the German Cancer Research Center as well as the participating researchers agree to: adhere to the rules of good scientific practice, have adhered to the guidelines regarding publication lists and bibliographies (cf. appendices I and II), observe all laws and regulations relevant to the research program and in particular to attain all necessary approvals, certifications, etc., in a timely manner, and - if applicable inform the DFG immediately if funding for this undertaking is requested from a third party. Proposals previously submitted to a third party and proposals involving major instrumentation must be mentioned in section 9.5 “Proposal submission to other funding organisations”, inform the DFG liaison officer of Heidelberg University about the proposal submission, plan and conduct any experiments involving humans, including identifiable samples taken from humans and identifiable data, in compliance with the most current versions of the German Embryo Protection Act (Embryonenschutzgesetz), Stem Cell Act (Stammzellgesetz), Pharmaceutical Drugs Act (Arzneimittelgesetz), Medical Devices Act (Medizinproduktegesetz), and the Declaration of Helsinki. plan and conduct any animal experiments in compliance with the Animal Protection Act (Tierschutzgesetz) and the Experimental Animals Ordinance (Versuchstierverordnung). adhere to the provisions of the Genetic Engineering Act (Gentechnikgesetz) with regard to experiments involving genetically modified organisms (GMOs). We accept the foregoing conditions and obligations. By accepting funding, the applicant university and the participating researchers agree to: a) use the grant exclusively and in a targeted manner to achieve the objectives of the Research Training Group as specified in the proposal; conform to the relevant regulations of the DFG in the use and accounting of funds; observe especially the usage guidelines for Research Training Groups (DFG form 2.22, in German); and not use the grant to finance core support. PROGRAM b) submit to the DFG progress reports on the Research Training Group according to the dates specified in the award letter; participate in the annual survey to evaluate the program; and present financial accounts to the DFG detailing the use of funds. 11 Signatures ___________________ Prof. Dr. Sergij Goerdt Designated Speaker _____________________ Prof. Dr. Martin Leverkus Designated Vice-Speaker ___________________ Prof. Dr. Otmar Wiestler German Cancer Research Center CEO _____________________ Prof. Dr. Bernhard Eitel University of Heidelberg Rector 50 Appendix I List of Published Research Relevant to the Research Program PROF. DR. PETER ANGEL 1. Briso E.M., J. Guinea-Viniegra, L. Bakiri, Z. Rogon, P. Petzelbauer, R. Eils, R. Wolf, M. Rincón, P. Angel, E.F. Wagner. 2013. Inflammation-mediated skin tumorigenesis induced by epidermal c-Fos. Genes Dev 27:1959-73. (IF 11,7) 2. Durchdewald M., J. Guinea-Viniegra, D. Haag, A. Riehl, P. Lichter, M. Hahn, E.F. Wagner, P. Angel*, J. Hess. 2008. Podoplanin is a novel Fos target gene in skin carcinogenesis. Cancer Res 68:6877-83. * corresponding author (IF 7,8) 3. Florin L., J. Knebel, P. Zigrino, B. Vonderstrass, C. Mauch, M. Schorpp-Kistner, A. Szabowski, P. Angel. 2006. Delayed wound healing and epidermal hyperproliferation in mice lacking JunB in the skin. J Inv Dermatol 126:902-911. (IF 6,3) 4. Gebhardt C., U. Breitenbach, J.P. Tuckermann, K.H. Richter, P. Angel. 2002. Calgranulins S100A8 and S100A9 are negatively regulated by glucocorticoids in a c-Fos-dependent manner and overexpressed throughout skin carcinogenesis. Oncogene 21:4266-76. (IF 6,3) 5. Gebhardt C., A. Riehl, M. Durchdewald, J. Németh, G. Fürstenberger, K. Müller-Decker, A. Enk, B. Arnold, A. Bierhaus, P.P. Nawroth, J. Hess, P. Angel. 2008. RAGE signaling sustaines inflammation and promotes tumor development; J Exp Med 205:275-85. (IF 13,8) 7. Klucky B., R. Mueller, I. Vogt, S. Teurich, B. Hartenstein, K. Breuhahn, C. Flechtenmacher, P. Angel *, J. Hess. 2007. The serine protease kallikrein 6 promotes keratinocyte proliferation and migration due to induction of E-cadherin shedding. Cancer Res 67:198-206. *corresponding author (IF 7,8) 8. Peterziel H., J. Müller, A. Danner, S. Barbus, H.K. Liu, B. Radlwimmer, T. Pietsch, P. Lichter, G. Schütz, J. Hess, P. Angel. 2012 Expression of podoplanin in human astrocytic brain tumors is controlled by the PI3K-AKT-AP-1 signaling pathway and promoter methylation. Neuro Oncol 14:426-39. (IF 5,7) 9. Tuckermann J., H. Reichardt, R. Arribaz, H. Richter, G. Schütz, P. Angel. 1999. The dimerization-independent function of the glucocorticoid receptor mediates repression of AP-1dependent gene expression in skin, J Cell Biol 147:1365-1370. (IF 10,2) 51 APPENDIX I 6. Hummerich L., R. Müller, J. Hess, F. Kokocinski, M. Hahn, G. Fürstenberger, C. Mauch, P. Lichter, P. Angel. 2006. Identification of novel tumour-associated genes differentially expressed in the process of squamous cell cancer development. Oncogene 25:111-21. (IF 6,3) PROF. DR. HELLMUT AUGUSTIN 1. Alajati A., A.M. Laib, H. Weber, A.M. Boos, A. Bartol, K. Ikenberg, T. Korff, H. Zentgraf, C. Obodozie, R. Graeser, S. Christian, G. Finkenzeller, G.B. Stark, M. Héroult, H.G. Augustin. 2008. Spheroid-based engineering of a human vasculature in mice. Nat Methods 5:439-45. (IF 23,57) 2. Felcht M., R. Luck, A. Schering, P. Seidel, K. Srivastava, J. Hu, A. Bartol, Y., Kienast, C. Vettel, E.K. Loos, S. Kutschera, S. Bartels, S. Appak, E. Besemfelder, D. Terhardt, E. Chavakis, T. Wieland, C. Klein, M. Thomas, A. Uemura, S. Goerdt, and H.G. Augustin. 2012. Angiopoietin2 differentially regulates angiogenesis through TIE2 and integrin signaling. J Clin Invest 122:1991-2005. (IF 12,81) 3. Fiedler U., Y. Reiss, M. Scharpfenecker, V. Grunow, S. Koidl, G. Thurston, N.W. Gale, M. Witzenrath, S. Rosseau, N. Suttorp, A. Sobke, M. Herrmann, K. Preissner, P. Vajkoczy, and H.G. Augustin. 2006. Angiopoietin-2 sensitizes endothelial cells to TNFα and plays a crucial role in the induction of inflammation. Nature Med 12:235-9. (IF 24,30) 4. Fiedler U., M. Scharpfenecker, S. Koidl, A. Hegen, V. Grunow, J.M. Schmidt, W. Kriz, G. Thurston, and H.G. Augustin. 2004. The Tie-2 ligand Angiopoietin-2 is stored in and rapidly released upon stimulation from endothelial cell Weibel-Palade bodies. Blood 103:4150-6. (IF 9,06) 5. Helfrich I., Scheffrahn, I., Bartling, S., Weis, J., von Felbert, V., Middleton, M., Kato, M., Ergün, S., Augustin, H. G.*, Schadendorf, D.*.2010. Resistance to antiangiogenic therapy is directed by vascular phenotype, vessel stabilization, and maturation in malignant melanoma. J Exp Med 207:491-503. (*equally contributing senior authors) (IF 13,21) 6. Hu J., Srivastava, K., Wieland, M., Runge, A., Mogler, C., Besemfelder, E., Terhardt, D., Vogel, M. J., Cao, L., Korn, C., Bartels, S., Thomas, M., and Augustin, H. G. .2014. Endothelial cellderived Angiopoietin-2 controls liver regeneration as a spatiotemporal rheostat. Science 343:416-9, 2014. (IF 31,03) APPENDIX I 7. Korn C., B. Scholz, J. Hu, K. Srivastava, J. Wojtarowicz, T. Arnsperger, R.H. Adams, M. Boutros*, H.G. Augustin*, I. Augustin*: 2014. Endothelial cell-derived non-canonical Wnt ligands control vascular pruning in angiogenesis. Development, in press. (*equal contribution) (IF 6,21) 8. Nasarre P., M. Thomas, K. Kruse, I. Helfrich, V. Wolter, C. Deppermann, D. Schadendorf, G. Thurston, U. Fiedler, and H.G. Augustin. 2009. Host-derived angiopoietin-2 affects early stages of tumor development and vessel maturation but is dispensable for later stages of tumor growth. Cancer Res 69:1324-33. (8,65) 9. Thomas M.*, M. Felcht*, K. Kruse, S. Kretschmer, C. Deppermann, A. Biesdorf, K. Rohr, A.V. Benest, U. Fiedler, H.G. Augustin. 2010. Angiopoietin-2 stimulation of endothelial cells induces alphavbeta3 integrin internalization and degradation. J Biol Chem 285:23842-9. (IF 4,65) * equal contribution 52 DR. IRIS AUGUSTIN 1. Augustin I., J. Gross, D. Baumann, C. Korn, G. Kerr, C. Mauch, W. Birchmeier, M. Boutros. 2013. Loss of epidermal Evi/Wls results in a phenotype resembling psoriasiform dermatitis. J Exp Med 210:1761-1777, 2013. (IF 13,21) 2. Augustin I., V. Goidts, A. Bongers, G. Kerr, G. Vollert, B. Radlwimmer, C. Hartmann, C. Herold-Mende, G. Reifenberger, A. von Deimling, M. Boutros. 2012. The Wnt secretion protein Evi/Gpr177 promotes glioma tumorigenesis. EMBO Mol Med 1:38-51. (IF 7,80) 3. Korn C., B. Scholz, J. Hu, K. Srivastava, J. Wojtarowicz, T. Arnsperger, R.H. Adams, M Boutros*, H.G. Augustin*, I. Augustin*. 2014. Endothelial cell-derived non-canonical Wnt ligands control vascular pruning in angiogenesis. Development, in press. (*equal contribution) (IF 6,21) APPENDIX I 4. Voloshanenko O., G. Erdmann , T.D. Dubash , I. Augustin , M. Metzig , G. Moffa , C. Hundsrucker, G. Kerr, T. Sandmann, B. Anchang, K. Demir, C. Boehm, S. Leible, C.R. Ball, H. Glimm, R. Spang, M. Boutros. 2013. Wnt secretion is required to maintain high levels of Wnt activity in colon cancer cells. Nat Commun 4:2610. (IF 10,02) 53 PROF. DR. MICHAEL BOUTROS 1. Augustin I., V. Goidts, A. Bongers, G. Kerr, G. Vollert, B. Radlwimmer, C. Hartmann, C. HeroldMende, G. Reifenberger, A. von Deimling, M. Boutros. 2012. The Wnt secretion protein Evi/Gpr177 promotes glioma tumourigenesis. EMBO Mol Med 4:38-51. (IF 7.80) 2. Augustin I., D. Baumann, D, C. Korn, G. Kerr, T. Grigoryan, C. Mauch, W. Birchmeier, M. Boutros. 2013. Loss of epidermal Evi/Wls results in a phenotype resembling psoriasiform dermatitis. J Exp Med. 210:1761-1777. (IF 13.21) 3. Bartscherer K., N. Pelte, D. Ingelfinger, M. Boutros. 2006. Secretion of Wnt ligands requires Evi, a conserved transmembrane protein. Cell 125:523-533. (IF 31.98) 4. Boutros M., J. Mihaly, T. Bouwmeester, and M. Mlodzik. 2000. Signaling specificity by Frizzled receptors in Drosophila. Science 288:1825-1828. (IF 31.03) 5. Boutros M., N. Paricio, D.I. Strutt, M. Mlodzik. 1998. Dishevelled activates JNK and discriminates between JNK pathways in planar polarity and wingless signaling. Cell 94:109118. (IF 31.96) 6. Cruciat C.M., B. Ohkawara, S.P. Acebron, E. Karaulanov, C. Reinhard, D. Ingelfinger, M. Boutros, C. Niehrs. 2010. Requirement of prorenin receptor and vacuolar H+-ATPasemediated acidification for Wnt signaling. Science 327:459-463. (IF 31.03) 7. Fuchs F., G. Pau, D. Kranz, O. Sklyar, C. Budjan, S. Steinbrink, T. Horn, A. Pedal, W. Huber, M. Boutros. 2010. Clustering phenotype populations by genome-wide RNAi and multiparametric imaging. Mol Syst Biol 6:370. (IF11.34) 8. Gross J.C., V. Chaudhary, K. Bartscherer, M. Boutros. 2012. Active Wnt proteins are secreted on exosomes. Nat Cell Biol 14:1036-1045. (IF 20.76) 9. Horn T., T. Sandmann, B. Fischer, E. Axelsson, W. Huber, M. Boutros. 2011. Mapping of signaling networks through synthetic genetic interaction analysis by RNAi. Nat Meth 8:341-346. (IF 23.57) APPENDIX I 54 PD DR. ADELHEID CERWENKA 1. Fiegler N., S. Textor, A. Arnold, A. Rölle, I. Oehme, K. Breuhahn, G. Moldenhauer, M. WitzensHarig, A. Cerwenka. 2013. Downregulation of the activating NKp30 ligand B7-H6 by HDAC inhibitors impairs tumor cell recognition by NK cells. Blood 122:684-93. (IF 9.1) 2. Ni J., M. Miller, A. Stojanovic, N. Garbi, A. Cerwenka. 2012. Sustained effector function of IL12/15/18 preactivated NK cells against established tumors. J Exp Med 209:2351-2365. (IF 13.2) 3. Schlecker E., A. Stojanovic, C. Eisen, C. Quack, C.S. Falk, V. Umansky, A. Cerwenka. 2012. Tumor-infiltrating monocytic myeloid-derived suppressor cells mediate CCR5-dependent recruitment of regulatory T cells favouring tumor growth. J.Immunol 189:5602-5611. (IF 5.5) 4. Textor S., N. Fiegler, A. Arnold, A. Porgador, T.G. Hofmann, A. Cerwenka. 2011. Human NK cells are alerted to induction of p53 in cancer cells by up-regulation of the NKG2D-ligands ULBP1 and ULBP2. Cancer Res 71:5998-6009. (IF 8.7) 5. Nausch N., I.E. Galani, E. Schlecker, A. Cerwenka. 2008. Mononuclear Myeloid-Derived “Suppressor” Cells express RAE-1 and activate NK cells. Blood 112:4080-9. (IF 9.1) 6. Wendel M., I.E. Galani, E. Suri-Payer, A. Cerwenka. 2008. NK cell accumulation in tumors is dependent on IFN-g and CXCR3 ligands. Cancer Res. 68:8437-45. (IF 8.7) 7. Cerwenka A., J.L. Baron, L. L. Lanier. 2001. Ectopic expression of retinoic acid early inducible1 gene (RAE-1) permits NK cell-mediated rejection of a MHC class I-bearing tumor in vivo. Proc Natl Acad Sci USA 98:11521-6. (IF 9.7) 8. Cerwenka A., L.L. Lanier. NK cells, viruses and cancer. 2001. Nat Immunol Rev 1:41-49. (IF 33.1) APPENDIX I 9. Cerwenka A., A.B.H. Bakker, T. McClanahan, J. Wagner, J. Wu, J.H. Phillips, L.L. Lanier. 2000. Retinoic acid early inducible genes define a ligand family for the activating NKG2D receptor in mice. Immunity 12:721-727. (IF 19.8) 55 PROF. DR. ALEXANDER H. ENK 1. Gebhardt C., A. Riehl, M. Durchdewald, J. Németh, G. Fürstenberger, K. Müller-Decker, A.H. Enk, B. Arnold, A. Bierhaus, P. Nawroth, J. Hess, P. Angel. 2008. RAGE signaling sustains inflammation and promotes tumor development. J Exp Med 205:275-285. (IF 13,21) 2. Jonuleit H., E. Schmitt, H. Kakirman, M. Stassen, J. Knop, A.H. Enk. 2002. Infectious tolerance: human CD25(+) regulatory T cells convey suppressor activity to conventional CD4(+) T helper cells. J Exp Med 196:255-60. (IF 13,21) 3. Jonuleit H., E. Schmitt, M. Stassen, A. Tuettenberg, J. Knop, A.H. Enk. 2001. Identification and functional characterization of human CD4(+)CD25(+) T cells with regulatory properties isolated from peripheral blood. J Exp Med 193:1285-94. (IF 13,21) 4. Lonsdorf A.S., S.T. Hwang, A.H. Enk. 2009. Chemokine receptors in T-cell-mediated diseases of the skin. J Invest Dermatol 129:2552-66. (IF 6,19) 5. Mahnke K, Y. Qian, S. Fondel, J. Brueck, C. Becker, A.H. Enk. 2005. Targeting of antigens to activated dendritic cells in vivo cures metastatic melanoma in mice. Cancer Res 65:7007-12. (IF 8,65) 6. Mahnke K., Y. Qian, J Knop, A.H. Enk. 2003. Dendritic cells, engineered to secrete a T-cell receptor mimic peptide, induce antigen-specific immunosuppression in vivo. Nat Biotechnol 21:903-8. (IF 32,43) 7. Mahnke K., Y. Qian, J. Knop, A.H. Enk. 2003. Induction of CD4+/CD25+ regulatory T cells by targeting of antigens to immature dendritic cells. Blood 101:4862-9. (IF 9,06) 8. Steinbrink K., E. Graulich, S. Kubsch, J. Knop, A.H. Enk. 2002. CD4(+) and CD8(+) anergic T cells induced by interleukin-10-treated human dendritic cells display antigen-specific suppressor activity. Blood 99:2468-76. (IF 9,06) APPENDIX I 9. Steinbrink K., H. Jonuleit, G. Müller, G. Schuler, J. Knop, A.H. Enk. 1999. Interleukin-10treated human dendritic cells induce a melanoma-antigen-specific anergy in CD8(+) T cells resulting in a failure to lyse tumor cells. Blood 93:1634-42. (IF 9,06) 56 DR. MORITZ FELCHT 1. Felcht M., R. Luck, A. Schering, P. Seidel, K. Srivastava, J. Hu, A. Bartol, Y., Kienast, C. Vettel, E.K. Loos, S. Kutschera, S. Bartels, S. Appak, E. Besemfelder, D. Terhardt, E. Chavakis, T. Wieland, C. Klein, M. Thomas, A. Uemura, S. Goerdt, H.G. Augustin. 2012. Angiopoietin-2 differentially regulates angiogenesis through TIE2 and integrin signaling. J Clin Invest 122:1991-2005. (IF 12,81) APPENDIX I 2. Thomas M.*, M. Felcht*, K. Kruse, S. Kretschmer, C. Deppermann, A.V. Benest, U. Fiedler, H.G. Augustin. 2010. Angiopoietin-2 stimulation of endothelial cells induces alphavbeta3 integrin internalization and degradation. J Biol Chem 285:23842-9. (IF 4,65); *equal contribution 57 PD DR. CYRILL GÉRAUD 1. Bioulac-Sage P., S. Lepreux, K. Schledzewski, G. Cubel, C. Géraud, S. Goerdt, C. Balabaud. 2010. Identification of liver sinusoidal endothelial cells in the human liver. Liver Int 30:773-6. (IF 3.87) 2. Evdokimov K., S. Biswas, M. Adrian, J. Weber, K. Schledzewski, M. Winkler, S. Goerdt, C. Géraud. 2013. Proteolytic cleavage of LEDA-1/PIANP by furin-like proprotein convertases precedes its plasma membrane localization. Biochem Biophys Res Commun 434: 22-27. (IF 2.40) 3. Géraud C., C. Mogler, A. Runge, K. Evdokimov, S. Lu, K. Schledzewski, B. Arnold, G. Hämmerling, P.S. Koch, K. Breuhahn, T. Longerich, A. Marx, C. Weiss, F. Damm, A. Schmieder, P. Schirmacher, H.G. Augustin, S. Goerdt. 2013. Endothelial transdifferentiation in hepatocellular carcinoma: loss of Stabilin-2 expression in peri-tumourous liver correlates with increased survival. Liver Int 33:1428-40. (IF 3.87) 4. Géraud C.*, K. Evdokimov*, B.K. Straub, W.K. Peitsch, A. Demory, Y. Dorflinger, K. Schledzewski, A. Schmieder, P. Schemmer, H.G. Augustin, P. Schirmacher, S. Goerdt. 2012. Unique cell type-specific junctional complexes in vascular endothelium of human and rat liver sinusoids. PLoS One 7:e34206. (* C.G. and E.K. contributed equally to this work) (IF 3.73) 5. Géraud C.*, K. Schledzewski*, A. Demory, D. Klein, M. Kaus, F. Peyre, C. Sticht, K. Evdokimov, S. Lu, A. Schmieder, S. Goerdt. 2010. Liver sinusoidal endothelium: a microenvironment-dependent differentiation program in rat including the novel junctional protein liver endothelial differentiation-associated protein-1. Hepatology. 52:313-26. (IF 12.00) 6. Klein D., A. Demory, F. Peyre, J. Kroll, C. Géraud, N. Ohnesorge, K. Schledzewski, B. Arnold, S. Goerdt. 2009. Wnt2 acts as an angiogenic growth factor for non-sinusoidal endothelial cells and inhibits expression of stanniocalcin-1. Angiogenesis 12:251-65. (IF 3.97) APPENDIX I 7. Schledzewski K.*, C. Géraud*, B. Arnold, S. Wang, H.J. Grone, T. Kempf, K.C. Wollert, B.K. Straub, P. Schirmacher, A. Demory, H. Schonhaber, A. Gratchev, L. Dietz, H.J. Thierse, J. Kzhyshkowska, S. Goerdt. 2011. Deficiency of liver sinusoidal scavenger receptors stabilin-1 and -2 in mice causes glomerulofibrotic nephropathy via impaired hepatic clearance of noxious blood factors. J Clin Invest 121:703-14. (* K.S. and C.G. contributed equally to this work) (IF 12.81) 58 DR. PETER GESERICK 1. Diessenbacher P., M. Hupe, M.R. Sprick, A. Kerstan, P. Geserick, T.L. Haas, T. Wachter, M. Neumann, H. Walczak, J. Silke, M. Leverkus. 2008. NF-kappaB inhibition reveals differential mechanisms of TNF versus TRAIL-induced apoptosis upstream or at the level of caspase-8 activation independent of cIAP2. J Invest Dermatol 128:1134-1147. (IF 6,19) 2. Feoktistova M.*, P. Geserick*, D. Panayotova-Dimitrova, M. Leverkus. 2012. Pick your poison: the Ripoptosome, a cell death platform regulating apoptosis and necrosis. Cell Cycle 11:460-7. (* equal contribution) (IF 5,24) 3. Feoktistova M.*, P. Geserick*, B. Kellert, D. Panayotova-Dimitrova, C. Langlais, M. Hupe, K. Cain, M. MacFarlain, G. Häcker, M.Leverkus. 2011. cIAPs block Ripoptosome formation, a RIP1/caspase 8 containing intracellular cell death complex differentially regulated by cFLIP isoforms. Mol Cell 43:449-63 (* equal contribution) (IF 15,28) 4. Geserick P., M. Hupe, M. Moulin, M. Leverkus. 2010. RIP-in CD95L-induced cell death: The control of alternative death receptors pathways by cIAPs. Cell Cycle. 9:2689-2691. (IF 5,24) 5. Geserick P., M. Hupe, M. Moulin, W.W. Wong, M. Feoktistova, B. Kellert, H. Gollnick, J. Silke, M. Leverkus. 2009. Cellular IAPs inhibit a cryptic CD95-induced cell death by limiting RIP1 kinase recruitment. J Cell Biol 187:1037-1054. (IF 10,82) 6. Geserick P., C. Drewniok, M. Hupe, T.L. Haas, P. Diessenbacher, M.R. Sprick, M.P. Schon, F. Henkler, H. Gollnick, H. Walczak, M. Leverkus. 2008. Suppression of cFLIP is sufficient to sensitize human melanoma cells to TRAIL or CD95L-mediated apoptosis. Oncogene 27:32113220. (IF 7,36) 8. Leverkus M, P. Diessenbacher, P. Geserick. 2008. FLIPing the coin? Death receptor-mediated signals during skin tumorigenesis. Exp Dermatol 17:614-622. (IF 3,57) 9. Panayotova-Dimitrova D., M. Feoktistova, M. Ploesser, B. Kellert, M. Hupe, S. Horn, R. Makarov, F. Jensen, S. Porubsky, A. Schmieder, A.C. Zenclussen, A. Marx, A. Kerstan, P. Geserick, Y.W. He, M. Leverkus. 2013. cFLIP Regulates Skin Homeostasis and Protects against TNF-Induced Keratinocyte Apoptosis. Cell Rep 5:397-408. (IF ?) 59 APPENDIX I 7. Kavuri S.M., P. Geserick, D. Berg, D.P. Dimitrova, M. Feoktistova, D. Siegmund, H. Gollnick, M. Neumann, H. Wajant, M. Leverkus. 2011. Cellular FLICE-inhibitory Protein (cFLIP) Isoforms Block CD95- and TRAIL Death Receptor-induced Gene Induction Irrespective of Processing of Caspase-8 or cFLIP inthe Death-inducing Signaling Complex. J Biol Chem 286:16631-16646. (IF 4,65) PROF. DR. SERGIJ GOERDT 1. Géraud C., K. Schledzewski, A. Demory, D. Klein, M. Kaus, F. Peyre, C. Sticht, K. Evdokimov, S. Lu, A. Schmieder, S. Goerdt. 2010. Liver sinusoidal endothelium: a microenvironmentdependent differentiation program in rat including the novel junctional protein liver endothelial differentiation-associated protein-1. Hepatology 52:313-26. (IF 12.00) 2. Géraud C., K. Evdokimov K, B. K. Straub, W. K. Peitsch, A. Demory, Y. Dörflinger, K. Schledzewski, A. Schmieder, P. Schemmer, H. G. Augustin, P. Schirmacher, S. Goerdt. 2012. Unique cell type-specific junctional complexes in vascular endothelium of human and rat liver sinusoids. PLoS ONE 7:e34206. (IF 3.73) 3. Géraud C., C. Mogler, A. Runge, K. Evdokimov, S. Lu, K. Schledzewski, B. Arnold, G. Hämmerling, P.S. Koch, K. Breuhahn, T. Longerich, A. Marx, C. Weiss, F. Damm, A. Schmieder, P. Schirmacher, H.G. Augustin, S. Goerdt. 2013. Endothelial transdifferentiation in hepatocellular carcinoma: loss of Stabilin-2 expression in peri-tumourous liver correlates with increased survival. Liver Int 33:1428-40. (IF 3.87) 4. Klein D., A. Demory, F. Peyre, J. Kroll, H.G. Augustin, W. Helfrich, J. Kzhyshkowska, K. Schledzewski, B. Arnold, S. Goerdt. 2008. Wnt2 acts as a cell type-specific, autocrine growth factor in rat hepatic sinusoidal endothelial cells cross-stimulating the VEGF pathway. Hepatolog 47:1018-31. (IF 12.00) 5. Klein D., A. Demory, F. Peyre, J. Kroll, C. Géraud, N. Ohnesorge, K. Schledzewski, B. Arnold, S. Goerdt. 2009. Wnt2 acts as an angiogenic growth factor for non-sinusoidal endothelial cells and inhibits expression of stanniocalcin-1. Angiogenesis 12:251-65. (IF 3.97) 6. Kzhyshkowska J., S. Mamidi, A. Gratchev, E. Kremmer, C. Schmuttermaier, L. Krusell, G. Haus, J. Utikal, K. Schledzewski, J. Scholtze, S. Goerdt. 2006. Novel stabilin-1 interacting chitinase-like protein (SI-CLP) is up-regulated in alternatively activated macrophages and secreted via lysosomal pathway. Blood 107:3221-8. (IF 9.06) APPENDIX I 7. Schledzewski K., M. Falkowski, G. Moldenhauer, P. Metharom, J. Kzhyshkowska, R. Ganss, A. Demory, B. Falkowska-Hansen, H. Kurzen, S. Ugurel, G. Geginat, B. Arnold, S. Goerdt. 2006. Lymphatic endothelium-specific hyaluronan receptor LYVE-1 is expressed by stabilin-1+, F4/80+, CD11b+ macrophages in malignant tumours and wound healing tissue in vivo and in bone marrow cultures in vitro: implications for the assessment of lymphangiogenesis. J Pathol 209:67-77. (IF 7.58) 8. Schledzewski K., C. Géraud, B. Arnold, S. Wang, H.J. Grone, T. Kempf, K.C. Wollert, B.K. Straub, P. Schirmacher, A. Demory, H. Schonhaber, A. Gratchev, L. Dietz, H.J. Thierse, J. Kzhyshkowska, S. Goerdt. 2011. Deficiency of liver sinusoidal scavenger receptors stabilin-1 and -2 in mice causes glomerulofibrotic nephropathy via impaired hepatic clearance of noxious blood factors. J Clin Invest 121:703-14. (IF 12.81) 9. Schmieder A., K. Schledzewski, J. Michel, J.P. Tuckermann, L. Tome, C. Sticht, C. Gkaniatsou, J.P. Nicolay, A. Demory, J. Faulhaber, J. Kzhyshkowska, C. Géraud, S. Goerdt. 2010. Synergistic activation by p38MAPK and glucocorticoid signaling mediates induction of M2-like tumor-associated macrophages expressing the novel CD20 homolog MS4A8A. Int J Cancer 129:122-32 (IF 6.20) 60 PROF. DR. MARTIN LEVERKUS 1. Diessenbacher P., M. Hupe, M.R. Sprick, A. Kerstan, P. Geserick, T.L. Haas, T. Wachter, M. Neumann, H. Walczak, J. Silke, M. Leverkus. 2008. NF-kappaB inhibition reveals differential mechanisms of TNF versus TRAIL-induced apoptosis upstream or at the level of caspase-8 activation independent of cIAP2. J Invest Dermatol 128:1134-1147. (IF 6,19) 2. Feoktistova M., P. Geserick, B. Kellert, D.P. Dimitrova, C. Langlais, M. Hupe, K. Cain, M. Macfarlane, G. Hacker, M. Leverkus. 2011. cIAPs block Ripoptosome formation, a RIP1/caspase-8 containing intracellular cell death complex differentially regulated by cFLIP isoforms. Mol Cell 43:449-463. (IF 15,28) 3. Geserick P., C. Drewniok, M. Hupe, T.L. Haas, P. Diessenbacher, M.R. Sprick, M.P. Schon, F. Henkler, H. Gollnick, H. Walczak, M. Leverkus. 2008. Suppression of cFLIP is sufficient to sensitize human melanoma cells to TRAIL or CD95L-mediated apoptosis. Oncogene 27:32113220. (IF 7,36) 4. Geserick P., M. Hupe, M. Moulin, W.W. Wong, M. Feoktistova, B. Kellert, H. Gollnick, J. Silke, M. Leverkus. 2009. Cellular IAPs inhibit a cryptic CD95-induced cell death by limiting RIP1 kinase recruitment. J Cell Biol 187:1037-1054. (IF 10,82) 5. Kavuri S.M., P. Geserick, D. Berg, D.P. Dimitrova, M. Feoktistova, D. Siegmund, H. Gollnick, M. Neumann, H. Wajant, M. Leverkus. 2011. Cellular FLICE-inhibitory Protein (cFLIP) Isoforms Block CD95- and TRAIL Death Receptor-induced Gene Induction Irrespective of Processing of Caspase-8 or cFLIP in the Death-inducing Signaling Complex. J Biol Chem 286:16631-16646. (IF 4,65) 6. Leverkus M., M. Neumann, T. Mengling, C.T. Rauch, E.B. Brocker, P.H. Krammer, H. Walczak. 2000a. Regulation of tumor necrosis factor-related apoptosis-inducing ligand sensitivity in primary and transformed human keratinocytes. Cancer Res 60:553-559. (IF 8,65) 8. Leverkus M., H. Walczak, A. McLellan, H.W. Fries, G. Terbeck, E.B. Brocker, E.Kampgen. 2000b. Maturation of dendritic cells leads to up-regulation of cellular FLICE- inhibitory protein and concomitant down-regulation of death ligand- mediated apoptosis. Blood 96:2628-2631. (IF 9,06) 9. Panayotova-Dimitrova D., M. Feoktistova, M. Ploesser, B. Kellert, M. Hupe, S. Horn, R. Makarov, F. Jensen, S. Porubsky, A. Schmieder, A.C. Zenclussen, A. Marx, A. Kerstan, P. Geserick, Y. W.He, M. Leverkus. 2013. cFLIP Regulates Skin Homeostasis and Protects against TNF-Induced Keratinocyte Apoptosis. Cell Rep 5:397-408. (IF ?) 61 APPENDIX I 7. Leverkus M., M.R. Sprick, T. Wachter, A. Denk, E.B. Brocker, H. Walczak, M. Neumann. 2003. TRAIL-induced apoptosis and gene induction in HaCaT keratinocytes: differential contribution of TRAIL receptors 1 and 2. J Invest Dermatol 121:149-155. (IF 6,19) DR. ANKE S. LONSDORF 1. Chien A.J.*, E.C. Moore*, A.S. Lonsdorf, R.M. Kulikauskas, B.G. Rothberg, A. J. Berger, M.B Major, S.T. Hwang, D. L. Rimm, R. T. Moon. 2009. Activated Wnt/beta-catenin signaling in melanoma is associated with decreased proliferation in patient tumors and a murine melanoma model. Proc Natl Acad Sci USA. 106:1193-8. (IF 9.74) *equal contribution, alphabetical order 2. Fang L*, A.S. Lonsdorf*, S.T. Hwang. 2008. Immunotherapy for advanced melanoma. J Invest Dermatol 128:2596-605. (IF 6,19) * equal contribution, alphabetical order 3. Hedrick M.N.*, A.S. Lonsdorf*, A.K. Shirakawa, C.C. Richard Lee, F. Liao, S.P. Singh, H.H. Zhang, A. Grinberg, P.E. Love, S.T. Hwang, J.M. Farber. 2009. CCR6 is required for IL-23induced psoriasis-like inflammation in mice. J Clin Invest 119:2317-2329. (IF 12,81) * equal contribution, alphabetical order 4. Huang V., A.S. Lonsdorf, L. Fang, T. Kakinuma, V.C. Lee, E. Cha, H. Zhang, K. Nagao, M. Zaleska, W.L. Olszewski, Hwang S.T. 2008. Cutting edge: rapid accumulation of epidermal CCL27 in skin-draining lymph nodes following topical application of a contact sensitizer recruits CCR10-expressing T cells. J Immunol 180:6462-6466. (IF 5,52) 5. Kakinuma T., H. Nadiminti, A.S. Lonsdorf, T. Murakami, B.A. Perez, H. Kobayashi, S.T. Hwang 2007. Small numbers of residual tumor cells at the site of primary inoculation are critical for anti-tumor immunity following challenge at a secondary location. Cancer Immunol Immunother 56:1119-31. (IF 3.63) 6. Langer H.F.*, V.V. Orlova*, C. Xie*, S. Kaul*, D. Schneider, A.S. Lonsdorf, M. Fahrleitner, E.Y. Choi, V. Dutoit, M. Pellegrini, S. Grossklaus, P.P. Nawroth, G. Baretton, S. Santoso, S.T. Hwang, B. Arnold, T. Chavakis. 2011. A novel function of Junctional Adhesion Molecule-C in mediating melanoma cell metastasis. Cancer Res 71:4096-4105. (IF 8,65) * equal contribution APPENDIX I 7. Lonsdorf A.S., B.F. Kraemer, M. Fahrleitner, T. Schoenberger, S. Gnerlich, S. Ring, S. Gehring, S.W. Schneider, M.J. Kruhlak, S.G. Meuth, B. Nieswandt, M. Gawaz, A.H. Enk, H.F. Langer. 2012. Engagement of αIIbβ3 (GPIIb/IIIa) with ανβ3 mediates interaction of melanoma cells with platelets - a connection to hematogeneous metastasis. J Biol Chem 287:2168-78. (IF 4,65) 8. Lonsdorf A.S., S.T. Hwang, A.H. Enk. 2009. Chemokine receptors in T-cell-mediated diseases of the skin. J Invest Dermatol 129:2552-2566. (IF 6,19) 9. Lonsdorf A.S., H. Kuekrek, B.V. Stern, B.O. Boehm, P.V. Lehmann, M. Tary Lehmann. 2003. Intratumor CpG-oligodeoxynucleotide injection induces protective antitumor T cell immunity. J Immunol 171:3941-3946. (IF 5,52) 62 PROF. DR. KNUT SCHÄKEL 1. Baumeister S.H., K. Holig, M. Bornhauser, M. Meurer, E.P. Rieber, K. Schäkel. 2007. G-CSF mobilizes slanDCs (6-sulfo LacNAc+ dendritic cells) with a high proinflammatory capacity. Blood 110:3078-3081. (IF 9,06) 2. Costantini C., F. Calzetti, O. Perbellini, A. Micheletti, C. Scarponi, S. Lonardi, M. Pelletier, K. Schäkel, G. Pizzolo, F. Facchetti, W. Vermi, C. Albanesi, M.A. Cassatella. 2011. Human neutrophils interact with both 6-sulfo LacNAc+ DC and NK cells to amplify NK-derived IFN{gamma}: role of CD18, ICAM-1, and ICAM-3. Blood 117:1677-1686. (IF 9,06) 3. Döbel T., A. Kunze, J. Babatz, K. Tränkner, A. Ludwig, M. Schmitz, A. Enk, K. Schäkel. 2013. FcγRIII (CD16) equips immature 6-sulfo LacNAc-expressing dendritic cells (slanDCs) with a unique capacity to handle IgG-complexed antigens. Blood 18:3609-18. (IF 9,06) 4. Hanse A., C. Gunther, J. Ingwersen, J. Starke, M. Schmitz, M. Bachmann, M. Meurer, E.P. Rieber, K. Schäkel. 2011. Human slan (6-sulfo LacNAc) dendritic cells are inflammatory dermal dendritic cells in psoriasis and drive strong TH17/TH1 T-cell responses. J Allergy Clin Immunol 127:787-794. (IF 12,05) 5. Randolph G.J., G. Sanchez-Schmitz, R.M. Liebman, K. Schäkel. 2002. The CD16(+) (FcgammaRIII(+)) subset of human monocytes preferentially becomes migratory dendritic cells in a model tissue setting. J Exp Med 196:517-527. (IF 13,22) 6. Schäkel K., R. Kannagi, B. Kniep, Y. Goto, C. Mitsuoka, J. Zwirner, A. Soruri, M. von Kietzell , E. Rieber. 2002. 6-Sulfo LacNAc, a novel carbohydrate modification of PSGL-1, defines an inflammatory type of human dendritic cells. Immunity 17:289-301. (IF 19,80) 8. Schäkel K., M. von Kietzell, A. Hansel, A. Ebling, L. Schulze, M. Haase, C. Semmler, M. Sarfati, A.N. Barclay, G.J. Randolph, M. Meurer, E.P. Rieber. 2006. Human 6-sulfo LacNAcexpressing dendritic cells are principal producers of early interleukin-12 and are controlled by erythrocytes. Immunity 24:767-777. (IF 19,80) 9. Schmitz M., S. Zhao, Y. Deuse, K. Schäkel, R. Wehner, H. Wohner, K. Holig, F. Wienforth, A. Kiessling, M. Bornhauser, A. Temme, M.A. Rieger, B. Weigle, M. Bachmann, E.P. Rieber. 2005. Tumoricidal potential of native blood dendritic cells: direct tumor cell killing and activation of NK cell-mediated cytotoxicity. J Immunol 174:4127-4134. (IF 5,52) 63 APPENDIX I 7. Schäkel K., E. Mayer, C. Federle, M. Schmitz, G. Riethmuller, E.P. Rieber. 1998. A novel dendritic cell population in human blood: one-step immunomagnetic isolation by a specific mAb (M-DC8) and in vitro priming of cytotoxic T lymphocytes. Eur J Immunol 28:4084-4093. (IF 4,97) DR. ASTRID SCHMIEDER 1. Michel J., K. Schonhaar, K. Schledzewski, C. Gkaniatsou, C. Sticht, B. Kellert, F. Lasitschka, C. Geraud, S. Goerdt, A. Schmieder. 2013. Identification of the novel differentiation marker MS4A8B and its murine homolog MS4A8A in colonic epithelial cells lost during neoplastic transformation in human colon. Cell Death Dis 4:e469. (IF 6.04) 2. Schmieder A., J. Michel, K. Schonhaar, S. Goerdt, K. Schledzewski. 2012a. Differentiation and gene expression profile of tumor-associated macrophages. Semin Cancer Biol 22:289-97. (IF 7,44) 3. Schmieder A., K. Schledzewski, J. Michel, K. Schonhaar, Y. Morias, T. Bosschaerts, J. Van den Bossche, P. Dorny, A. Sauer, C. Sticht, C. Geraud, Z. Waibler, A. Beschin, S. Goerdt. 2012b. The CD20 homolog Ms4a8a integrates pro- and anti-inflammatory signals in novel M2like macrophages and is expressed in parasite infection. Eur J Immunol 42:2971-82. (IF 4.97) 4. Schmieder A., K. Schledzewski, J. Michel, J.P. Tuckermann, L. Tome, C. Sticht, C. Gkaniatsou, J.P. Nicolay, A. Demory, J. Faulhaber, J. Kzhyshkowska, C. Geraud, S. Goerdt. 2011. Synergistic activation by p38MAPK and glucocorticoid signaling mediates induction of M2-like tumor-associated macrophages expressing the novel CD20 homolog MS4A8A. Int J Cancer 129:122-32. (IF 6.20) 5. Schoenhaar K, Schledzewski K, Michel J, Dollt C, Gkaniatsou C, Géraud C, Kzhyshkowska J, Goerdt S, Schmieder A. 2013. Expression of Stabilin-1 in M2 macrophages in human granulomatous disease and melanocytic lesions. Int J Clin Exper Pathol, in press. (IF 2.2) APPENDIX I 64 PROF. DR. STEFAN W. SCHNEIDER 1. Bauer A.T, E.A. Strozyk, C. Gorzelanny, C. Westerhausen, A. Desch, M.F. Schneider, S. W. Schneider. 2011. Cytotoxicity of silica nanoparticles through exocytosis of von Willebrand factor and necrotic cell death in primary human endothelial cells. Biomaterials 32:8385-93. (IF 7,60) 2. Chen H., M.A. Fallah, V. Huck, J.I. Angerer, A.J. Reininger, S.W. Schneider, M.F. Schneider, A. Alexander-Katz. 2013. Blood-clotting-inspired reversible polymer-colloid composite assembly in flow. Nat Commun 4:1333. (IF 10,02) 3. Desch A., E.A. Strozyk, A.T. Bauer, V. Niemeyer, T. Wieland, S.W. Schneider. 2012. Highly invasive melanoma cells activate the vascular endothelium via a MMP-2/ integrin ανβ5-induced secretion of VEGF-A. Am J Pathology 181:693-705. (IF 4.51) 4. Görge T., A. Barg, E.M. Schnäker, B. Pöppelmann, V. Shpacovitch, A. Rattenholl, T.A. Luger, M. Steinhoff, S.W. Schneider. 2006. Melanoma-derived MMP-1 targets endothelial PAR1 promoting endothelial cell activation. Cancer Res 66:7766-74. (IF 8,65) 5. Kerk N., E.A. Strozyk, B. Pöppelmann, S.W. Schneider. 2010. The mechanism of melanomaassociated thrombin activity and von Willebrand factor release from endothelial cells. J Invest Dermatol 130:2259-68. (IF 6,19) 6. Pappelbaum K.I., C. Gorzelanny, S. Grässle, J. Suckau M.W. Laschke, M. Bischoff, C. Bauer, M. Schorpp-Kistner, C. Weidenmaier, R. Schneppenheim, T. Obser, B. Sinha, S.W. Schneider. 2013. Ultra-large von Willebrand factor fibers mediate luminal Staphylococcus aureus adhesion to an intact endothelial cell layer under shear stress. Circulation 128:50-59. (IF 15,20) 8. Schneider S. W., S. Nuschele, A. Wixforth, A. Alexander-Katz, R.R. Netz, C. Gorzelanny, M.F. Schneider. 2007. Shear-induced unfolding triggers adhesion of VWF fibers. Pro. Natl Acad Sci USA 104:7899-903. (IF 9,74) 9. Steinhoff M., A. Steinhoff, B. Homey, T.A. Luger, S.W.Schneider. 2006. Role of vasculature in atopic dermatitis. J Allergy Clin Immunol. 118:190-7. (IF 12,05) 65 APPENDIX I 7. Riehemann K, S.W. Schneider, T.A. Luger, B. Godin, M. Ferrari, H. Fuchs. 2009. Nanomedicine: challenge and perspectives. Angewandte Chemie Int. edition 48:872-97. (IF 13,73) PROF. DR. JONATHAN SLEEMAN 1. Baumann P., N. Cremers, F. Kroese, G. Orend, R. Chiquet-Ehrismann, T. Uede, H. Yagita, J. P. Sleeman. 2005. CD24 expression causes the acquisition of multiple cellular properties associated with tumor growth and metastasis. Cancer Res 65:10783-10793. (IF 8.65) 2. Baumann P., W. Thiele, N. Cremers, S. Muppala, J. Krachulec, M. Diefenbacher, O. Kassel, G. Mudduluru, H. Allgayer, M. Frame, J.P. Sleeman. 2012. CD24 interacts with and promotes the activity of c-src within lipid rafts in breast cancer cells, thereby increasing integrin-dependent adhesion. Cell Mol Life Sci, 69:435-448. (IF 5.62) 3. Krishnan J., V. Kirkin, A. Steffen, M. Hegen, D. Weih, S. Tomarev, J. Wilting, J. P. Sleeman. 2003. Differential in vivo and in vitro expression of VEGF-C and VEGF-D in tumors and its relationship to lymphatic metastasis in immunocompetent rats. Cancer Res 63:713-722. (IF 8.65) 4. Kuch V., C. Schreiber, W. Thiele, V. Umansky, J. P. Sleeman. 2013. Tumor initiating properties of breast cancer and melanoma cells in vivo are not invariably reflected by spheroid formation in vitro, but can be increased by long-term culturing as adherent monolayers. Int J Cancer 132:E94-105. (IF 6.20) 5. Müller T., U. Stein, A. Poletti, L. Garzia, M. Rothley, D. Plaumann, W. Thiele, M. Bauer, A. Galasso, P. Schlag, M. Pankratz, M. Zollo, J. P. Sleeman. 2010. ASAP1 promotes tumor cell motility and invasiveness, stimulates metastasis formation in vivo, and correlates with poor survival in colorectal cancer patients. Oncogene 29:2393-2403. (IF 7.36) 6. Neeb A., S. Wallbaum, N. Novac, I. Scholl, S. Dukovic-Schulze, C. Schreiber, P. Schlag, J. Moll, U. Stein, J. P. Sleeman. 2012. The immediate early gene Ier2 promotes tumor cell motility and metastasis, and predicts poor survival of colorectal carcinoma patients. Oncogene 31:3796-806. (IF 7.36) 7. Nestl A., O. Von Stein, K. Zatloukal, W. G. Thies, P. Herrlich, M. Hofmann, J. P. Sleeman. 2001. Gene expression patterns associated with the metastatic phenotype in rodent and human tumors. Cancer Res. 61:1569-1577. (IF 8.65) APPENDIX I 8. Schreiber C, V. Kuch, V. Umansky, J. P. Sleeman. 2013. Autochthonous mouse melanoma and mammary tumors do not express the pluripotency genes Oct4 and Nanog. PLoS One 8:e57465. (IF 3.73) 9. Thiele W., N. Novac, S. Mink, C. Schreiber, D. Plaumann, J. Fritzmann, C. Schwager, T. Regiert, P. E. Huber, U. Stein, P. Schlag, J. Moll, A. Abdollahi, J.P. Sleeman. 2011. Discovery of a novel tumor metastasis-promoting gene NVM-1. J Pathol 225:96-105. (IF 7.59) 66 PROF. DR. VIKTOR UMANSKY 1. Jayaraman P., F. Parikh, E. Lopez-Rivera, Y. Hailemichael, A. Clark, G. Ma, D. Cannan, M. Ramacher, M. Kato, W.W. Overwijk, S.-H. Chen, V. Umansky, A.G. Sikora. 2012. Inducible nitric oxide synthase (iNOS) controls induction of functional myeloid derived suppressor cells (MDSC). J Immunol 188:5365-5376. (IF 5.52) 2. Meyer C., A. Sevko, M. Ramacher, A.V. Bazhin, C.S. Falk, W. Osen, I. Borrello, M. Kato, D. Schadendorf, M. Baniyash, V. Umansky. 2011. Chronic inflammation promotes myeloid derived suppressor cell activation blocking antitumor immunity in transgenic mouse melanoma model. Proc Natl Acad Sci USA 108:17111-17116. (IF 9.74) 3. Schlecker E., A. Stojanovic, C. Eisen, C. Quack, C.S. Falk, V. Umansky, A. Cerwenka, A. 2012. Tumor-Infiltrating Monocytic Myeloid-Derived Suppressor Cells Mediate CCR5Dependent Recruitment of Regulatory T Cells Favoring Tumor Growth. J. Immunol 189:56025611. (IF 5.52) 4. Sevko A., M. Sade-Feldman, J. Kanterman, T. Michels, C.S. Falk, L. Umansky, M. Ramacher, M. Kato, D. Schadendorf, M. Baniyash, V. Umansky. 2013. Low dose cyclophosphamideenhanced chronic inflammation prevents anti-tumor effects in transgenic mouse melanoma model. J Invest Dermatol 133:1610-1619. (IF 6.19) 5. Sevko A., T. Michels, M. Vrohlings, L. Umansky, P. Beckhove, M. Kato, G.V. Shurin, M.R. Shurin, V. Umansky. 2013. Anti-tumor effect of paclitaxel is mediated by inhibition of MDSCs and chronic inflammation in the spontaneous melanoma model. J Immunol 190:2464-2471. (IF 5.52). 6. Umansky V., O. Abschuetz, W. Osen, M. Ramacher, F. Zhao, M. Kato, D. Schadendorf. 2008. Melanoma specific memory T cells are functionally active in ret transgenic mice without macroscopical tumors. Cancer Res 68:9451-9458. (IF 8.65) 8. Zhao F., C. Falk, W. Osen, M. Kato, D. Schadendorf, V. Umansky. 2009. Activation of p38 MAPK Drives Dendritic Cells to Become Tolerogenic in Ret Transgenic Mice Spontaneously Developing Melanoma. Clin Cancer Res 15:4382-4390. (IF 7.84) 67 APPENDIX I 7. Umansky V., A. Sevko. 2012. Melanoma-induced immunosuppression and its neutralization. Semin. Cancer Biol 22:319-326. (IF 7.44) PROF. DR. JOCHEN UTIKAL 1. Bernhardt M., E. Orouji, L. Larribere, C. Gebhardt, J. Utikal. 2014. Efficacy of vemurafenib in a trametinib resistant stage IV melanoma patient. Clin Cancer Res (in press) (IF 7,83) 2. Eminli S*, J. Utikal *, K. Arnold, R. Jaenisch, K. Hochedlinger. 2008. Reprogramming of neural progenitor cells into induced pluripotent stem cells in the absence of exogenous Sox2 expression. . Stem Cells 26:2467-74. * authors contributed equally (IF 7,70) 3. Flaherty K.T., C. Robert, P. Hersey, P. Nathan, C. Garbe, M. Milhem, L.V. Demidov, J.C. Hassel, P. Rutkowski, P. Mohr, R. Dummer, U. Trefzer, J.M. Larkin, J. Utikal, B. Dreno, M. Nyakas, M.R. Middleton, J.C. Becker, M. Casey, L.J. Sherman, F.S. Wu, D. Ouellet, A.M. Martin, K. Patel, D. Schadendorf; the METRIC Study Group. 2012. Improved Survival with MEK Inhibition in BRAF-Mutated Melanoma. N Engl J Med 367:107-14. (IF 51,65) 4. Hirata A*, J. Utikal *, S. Yamashita, H. Aoki, A. Watanabe, T. Yamamoto, H. Okano, N. Bardeesy, T. Kunisada, T. Ushijima, A. Hara, R. Jaenisch, K. Hochedlinger, Y. Yamada. 2013. Dose-dependent roles for canonical Wnt signalling in de novo crypt formation and cell cycle properties of the colonic epithelium. Development 140:66-75. *authors contributed equally (IF 6,20) 5. Maherali N., T. Ahfeldt, A. Rigamonte, J. Utikal, C. Cowen, K. Hochedlinger. 2008. A highefficiency system for the generation and study of human induced pluripotent stem cells. Cell Stem Cell 3:340-5. (IF 25,31) 6. Maherali N, R. Sridharan, W. Xie, J. Utikal, S. Eminli, K. Arnold, M. Stadtfeld, R. Yachechko, J. Tchieu, R. Jaenisch, K. Plath, K. Hochedlinger. 2007. Directly reprogrammed fibroblasts show global epigenetic remodeling and widespread tissue contribution. Cell Stem Cell 1:55-70. (IF 25,31) 7. Stadtfeld M., M. Nagaya, J. Utikal, G. Weir, K. Hochedlinger. 2008. Induced pluripotent stem cells generated without viral integration. Science 322:945-9. (IF 31,02) APPENDIX I 8. Utikal J., J.M. Polo, M. Stadtfeld, N. Maherali, W. Kulalert, R.M. Walsh, A. Khalil, J.G. Rheinwald, K. Hochedlinger. 2009. Immortalization eliminates a roadblock during the reprogramming of somatic cells into iPS cells. Nature 460:1145-8. (IF 38,59) 9. Utikal J., N. Maherali, W. Kulalert, K. Hochedlinger. 2009. Sox2 is dispensable for the reprogramming of melanocytes and melanoma cells into induced pluripotent stem cells. J Cell Sci 122:3502-10. (IF 5,87) 68 PROF. DR. FRANK WINKLER 1. Egea V, L. von Baumgarten, C. Schichor, B. Berninger, T. Popp, P. Neth, R. Goldbrunner, Y. Kienast, F. Winkler, M. Jochum, C. Ries. 2011. TNF-alpha respecifies human mesenchymal stem cells to a neural fate and promotes migration toward experimental glioma. Cell Death Differ 18:853-63. (IF 8.8) 2. Garkavtsev I., S.V. Kozin, O. Chernova, L. Xu, F. Winkler, E. Brown, G.H. Barnett, R.K. Jain. 2004. The candidate tumour suppressor protein ING4 regulates brain tumour growth and angiogenesis. Nature 428:328-32. (IF 36.3) 3. Kienast Y., L. von Baumgarten, M. Fuhrmann, W. Klinkert, R. Goldbrunner, J. Herms, F. Winkler. 2010. Real-time imaging reveals the single steps of brain metastasis formation. Nature Medicine 16:116-122. (IF 22.5) 4. Tong R.T., Y. Boucher, S.V. Kozin, F. Winkler, D.J. Hicklin, R.K. Jain. 2004. Vascular normalization by vascular endothelial growth factor receptor 2 blockade induces a pressure gradient across the vasculature and improves drug penetration in tumors. Cancer Res 64:3731-6. (IF 7.9) 5. von Baumgarten L., D. Brucker, A. Tirniceru, Y. Kienast, S. Grau, S. Burgold, J. Herms, F. Winkler. 2011. Bevacizumab has differential and dose-dependent effects on glioma blood vessels and tumor cells. Clin Cancer Res 17:6192-205. (IF 7.7) 6. Winkler F., Y. Kienast, M. Fuhrmann, L. von Baumgarten, S. Burgold, G. Mitteregger, J. Herms. 2009. Imaging glioma cell invasion in vivo reveals mechanisms of dissemination and peritumoral angiogenesis. Glia 57:1306-15. (IF 4.8) 8. Xu L., D. M. Cochran, R. T. Tong, F. Winkler, S. Kashiwagi, R. K. Jain, D. Fukumura. 2006. Placenta growth factor overexpression inhibits tumor growth, angiogenesis, and metastasis by depleting vascular endothelial growth factor homodimers in orthotopic mouse models. Cancer Res 66:3971-3977. (IF 7.9) 69 APPENDIX I 7. Winkler F., S.V. Kozin, R.T. Tong, S. Chae, M.F. Booth, I. Garkavtsev, L. Xu, D.K. Hicklin, D. Fukumura, E. di Tomaso, L.L. Munn, R.K. Jain. 2004. Kinetics of vascular normalization by VEGFR2 blockade governs brain tumor response to radiation: Role of oxygenation, Angiopoietin-1, and matrix metalloproteinases. Cancer Cell 6:553-563. (IF 26.6) Appendix II 1 Biographical Sketches of the Participating Researchers PROF. DR. RER. NAT. PETER ANGEL Head of Division Signal Transduction and Growth Control (A100) German Cancer Research Center (DKFZ) Im Neuenheimer Feld 280 69120 Heidelberg +49-6221-42-4570 (Fon) +49-6221-42-4554 (Fax) [email protected] Curriculum vitae 1995 - to date 1990 - 1995 1987 - 1989 1993 - 1987 1982 - 1993 Head of Division "Signal Transduction and Growth Control" at the German Cancer Research Center (DKFZ), Heidelberg Head of Research Group at the Institute of Genetics, Research Center Karlsruhe Postdoctoral Fellow at the University of California San Diego PhD Study at the University Karlsuhe (summa cum laude) Diploma at the University Karlsruhe (excellent) APPENDIX II Awards and Appointments January 1990 Young Investor Award (Heisenberg Stipendium) of the German Research Society (DFG) November 2003 appointed Full Professor in "Molecular Cell Biology: signal Transduction and Growth Control" at University Heidelberg 2002 - 2003 Coordinator of Research Program "Tumor Cell Biology" of DKFZ 2003 - 2006 Dep. Coordinator of Res. Program "Cell and Tumor Biology" of DKFZ 2004 - 2012 Elected member of the Study Section "Cell Biology" of the DFG since 2005 Elected Member of the Board of Trustees (Kuratorium) of the DKFZ since 2008 Elected EMBO Member since 2011 Elected member of the Board of Directors of the DKFZ-ZMBH Alliance since 2012 National Coordinator of the BMBF-funded Program of German-Israeli Cooperation in Cancer Research Areas of research expertise Signal transduction, transcription factors and genetic programs, communication, genetically modified mouse models, tumor biology Inflammation, cell-cell 5 selected (most important) publications 1. Gebhardt C, Riehl A, Durchdewald M, Németh J, Fürstenberger G, Müller-Decker K, Enk A, Arnold B, Bierhaus A, Nawroth PP, Hess J, Angel P. (2008) RAGE signaling sustaines inflammation and promotes tumor development; J Exp Med 205:275-85. (IF 13,8) 2. Zenz R, Eferl R, Kenner L, Florin L, Hummerich Mehic D, Scheuch H, Angel P, Tschachler E, Wagner EF. (2005) Psoriasis-like skin disease and arthritis caused by inducible epidermal deletion of Jun proteins. Nature 437:369-75. (IF 36,3) 70 3. Szabowski A, Maas-Szabowski N, Andrecht S, Kolbus A, Schorpp-Kistner M, Fusenig NE, Angel P. (2000) c-Jun and JunB antagonistically control cytokine-regulated mesenchymalepidermal interaction in skin. Cell 103:745-75. (IF 32,4) 4. Angel P, Allegretto EA, Okino ST, Hattori K, Boyle WJ, Hunter T, Karin M. (1988) Oncogene Jun encodes a sequence specific trans-activator similar to AP1. Nature 332:166-171. (IF 36,3) 5. Angel P, Imagawa M, Chiu R, Stein B, Imbra RJ, Rahmsdorf HJ, Jonat C, Herrlich P and Karin M. (1987) Phorbol ester inducible genes contain a common cis element recognized by a TPA inducible trans-acting factor. Cell 49:729-739. (IF 32,4) PhD/MD students (last 5 years) and titles of their theses PHD Jurisch-Yaksi, Nathalie (2005 - 09) Positive and negative regulator JunB: impact on chromatin remodeling and stress response. University Heidelberg. Final degree: magna cum laude Durchdewald, Moritz (2006 - 09) Identification of the Fos/AP-1-dependent genetic network implicated in epithelial carcinogenesis. University Heidelberg. Final degree: magna cum laude Nemeth, Julia: (2005 - 09) Function and regulation of S100 proteins in inflammation-associated carcinogenesis. University Heidelberg. Final degree: magna cum laude Riehl, Astrid: (2005 - 09) Identification and characterization of gene regulatory networks controlled by the receptor RAGE in inflammation and cancer. University Heidelberg, Final degree: summa cum laude Hildenbrand, Maike (2006 - 10) Transcriptional regulation of the aspartic protease Taps and its function during cutaneous wound healing. University Heidelberg. Final degree: magna cum laude Krenzer, Stefanie (2007 - 10) Regulation and function of the kallikrein-related peptidase 6 in the development of malignant melanoma. University Heidelberg. Final degree: magna cum laude Wiechert, Lars (2009 - 12) Function of epithelial-derived S100a8 and S100 a9 proteins in tissue homeostasis and inflammation in transgenic mouse models. University Heidelberg. Final degree: magna cum laude Leibold, Julia (2010-2012: Function of the Receptor for Advanced Glycation End Products in Keratinocytes in the regulation of skin inflammation. University Heidelberg. Final degree: magna cum laude Schumacher, Marion (2010-2013) JNK-dependent dermal genetic program controls interdependent keratinocyte-fibroblast crosstalk promoting keratinocyte differentiation during cutaneous wound healing. University Heidelberg. Final degree: magna cum laude Current extramural funding DFG Transregio-SFB-77 Leberkrebs von der molekularen Pathogenese zur zielgerichteten Therapie (2010-2014): TP A07 "Regulation and Function of the S100A8/S100A9 protein complex in inflammation-associated liver carcinogenesis" DFG Transregio-SFB-23 "Vaskuläre Differenzierung und Remodellierung" (2013-2017) Teilprojekt B02 Role of JunB and JunB target genes in endothelial cell function and tumor angiogenesis" BMBF Programm AGENET (2011-2014): The role of AP-1 subunits in keratinocytes and cells of the dermal compartment for trans-regulatory mechanisms controlling tissue homeostasis and remodeling in the skin DKFZ-Ministry of Science and Technlogy (MOST) of Israel (2011-2014) S100-Rage signalling in liver tumour angiogenesis Helmholtz-Gemeinschaft: Pre-clinical Comprehensive Cancer Center (PCCC; 2013-2016) The role of podoplanin in in vivo mouse models of brain and gastrointestinal tumors 71 APPENDIX II Kiesow, Kristin (2009-12: miR-182 -anovel Junb target and regulator of lymphangiogenesis in Danio rerio. University Heidelberg. Final degree: magna cum laude PROF. DR. MED. VET. HELLMUT G. AUGUSTIN, PHD Professor and Director Joint Research Division Vascular Biology, Medical Faculty Mannheim (CBTM), Heidelberg University, and German Cancer Research Center Heidelberg (DKFZ-ZMBH-Alliance) Address MA: Section of Vascular Biology and Tumor Angiogenesis Center for Biomedicine and Medical Technology (CBTM) Medical Faculty Mannheim, University of Heidelberg Ludolf-Krehl-Straße 13-17 68167 Mannheim 0621-383-9962 (Fon) Address HD: Dept. of Vascular Oncology and Metastasis German Cancer Research Center (DKFZ) Im Neuenheimer Feld 280 69120 Heidelberg 06221-42-1500 (Fon) [email protected] www.angiolab.de Curriculum vitae 3/2013-pres Director, Helmholtz Alliance “Preclinical Comprehensive Cancer Center” (PCCC, www.helmholtz-pccc.de) APPENDIX II 1/2011-pres. Deputy Director, Center for Biomedicine and Medical Technology Mannheim (CBTM), Medical Faculty Mannheim, Heidelberg University, Germany 1/2011-pres. Director, Center for Molecular Biology and German Cancer Research Center Alliance (DKFZ-ZMBH-Alliance) 1/2011-pres. Speaker, Cell and Tumor Biology Research Program (FSP-A), German Cancer Research Center, Heidelberg, Germany 7/2009-pres. Speaker of the SFB-TR23 “Vascular Differentiation and Remodeling” of the Universities Heidelberg and Frankfurt (www.transregio23.de) 5/2006-pres. Aventis Foundation-endowed Chair for Vascular Biology and Angiogenesis Research, Joint Research Division Vascular Biology, Medical Faculty Mannheim (CBTM), Heidelberg University, and German Cancer Research Center Heidelberg (DKFZ-ZMBH Alliance), Germany 5/2005-pres. Founding member and elected Chairman of VWFB e.V. (Verein zur Förderung wissenschaftlicher Fachtagungen e.v.; www.vwfb.de) 5/2005-pres. Coordinator of the nationwide German tumor-vessel interaction Priority Research Grant (SPP1190, www.tumorvessel.de) 7/2005-6/2009 Vice Speaker of the SFB-TR23 “Vascular Differentiation and Remodeling” of the Universities Frankfurt, Heidelberg, and Freiburg (www.transregio23.de) 2002-2006 Adjunct Professor, Medical Faculty of the Albert-Ludwigs-University Freiburg, Germany 2001-2006 Head, Dept. of Vascular Biology & Angiogenesis Research, Institute of Molecular Oncology, Tumor Biology Center, Freiburg, Germany (private, non-academic research institute) 72 9/1998- 5/2006 Coordinator of nationwide German angiogenesis Priority Research Grant (SPP1069, www.angiogenese.de) 1992-2001 Research Assistant Professor (C1, C2), Clinic for Gynecology and Obstetrics, University of Göttingen, Germany Nov. 5, 1997 Venia legendi (Habilitation) in Molecular Cell Biology, University of Göttingen, Germany Aug. 24, 1992 PhD, Cornell University, Ithaca, NY, USA 1988-1992 Graduate Student, Dept. of Pathology, Cornell University, Ithaca, NY, USA Dec. 12, 1987 Doctoral degree Dr. med. vet. Dec. 3, 1984 License to practice 2/1997 Founder of the German vascular biology network (with biannual meeting series) 1984-1987 Residency and graduate training in Veterinary Pathology, School of Veterinary Medicine Hannover, Germany July 12, 1984 DVM, School of Veterinary Medicine Hannover, Germany 1980-1984 Veterinary Medicine, School of Veterinary Medicine Hannover, Germany 1978-1980 Physikum (B.S.), School of Veterinary Medicine Hannover, Germany Areas of research expertise The lab studies 1.) the molecular mechanisms of tumor angiogenesis focusing on angiogenesis regulating receptor tyrosine kinases, most notably on the Angiopoietin-Tie ligand-receptor system, 2.) the molecular mechanisms of physiological blood and lymphatic vessel formation, assembly, and maturation focusing on selected novel candidate molecules, 3.) the molecular mechanisms of tumor progression focusing on tumor-vessel interactions during metastasis (role of tumor cell – endothelial cell interactions in the control of site-specific metastasis), and 4.) translational tumor angiogenesis experiments aimed at defining the therapeutic window of anti-angiogenic tumor therapies. Conceptually, the lab’s work is considered as basic tumor biology research with the aim of identifying and validating novel therapeutic targets. Hu, J., Srivastava, K., Wieland, M., Runge, A., Mogler, C., Besemfelder, E., Terhardt, D., Vogel, M. J., Cao, L., Korn, C., Bartels, S., Thomas, M., and Augustin, H. G. (2014) Endothelial cell-derived Angiopoietin-2 controls liver regeneration as a spatiotemporal rheostat. Science, 343: 416-9, 2014 (IF 31,03) Felcht, M., R. Luck, A. Schering, P. Seidel, K. Srivastava, J. Hu, A. Bartol, Y., Kienast, C. Vettel, E.K. Loos, S. Kutschera, S. Bartels, S. Appak, E. Besemfelder, D. Terhardt, E. Chavakis, T. Wieland, C. Klein, M. Thomas, A. Uemura, S. Goerdt, and H.G. Augustin. (2012) Angiopoietin-2 differentially regulates angiogenesis through TIE2 and integrin signaling. J Clin Invest. 122:19912005 (IF 12,81) Helfrich, I., Scheffrahn, I., Bartling, S., Weis, J., von Felbert, V., Middleton, M., Kato, M., Ergün, S., Augustin, H. G.*, Schadendorf, D.* (2010) Resistance to antiangiogenic therapy is directed by vascular phenotype, vessel stabilization, and maturation in malignant melanoma. J Exp Med. 207: 491-503 (*equally contributing senior authors) (IF 13,21) Alajati, A., A.M. Laib, H. Weber, A.M. Boos, A. Bartol, K. Ikenberg, T. Korff, H. Zentgraf, C. Obodozie, R. Graeser, S. Christian, G. Finkenzeller, G.B. Stark, M. Héroult, and H.G. Augustin. 2008. Spheroid-based engineering of a human vasculature in mice. Nat Methods, 5: 439-45. (IF 23,57) Fiedler, U., Y. Reiss, M. Scharpfenecker, V. Grunow, S. Koidl, G. Thurston, N.W. Gale, M. Witzenrath, S. Rosseau, N. Suttorp, A. Sobke, M. Herrmann, K. Preissner, P. Vajkoczy, and H.G. Augustin. 2006. Angiopoietin-2 sensitizes endothelial cells to TNFα and plays a crucial role in the induction of inflammation. Nature Med. 12:235-9. (IF 24,30) 73 APPENDIX II 5 selected (most important) publications PhD students (last 5 years) and titles of their theses Markus Thomas, Molecular mechanisms of angiopoietin-2-mediated destabilization of the vascular endothelium, 2008. Karoline Kruse, Regulation der vaskulären Homöostase durch den Tie2 Liganden Angiopoietin-2 und Nitritoxid, 2008. Renate Becker, Molecular analysis of Endosialin and its interaction with Mac-2 Binding protein, 2009. Daniel Epting, Analysis of G-protein signaling molecules during vertebrate development and angiogenesis, 2009. Silke Kaltenthaler, Identification of CD36 as a novel regulator of lymphatic endothelial cell function, 2009. Anna Laib, Establishment of a novel human in vivo lymphangiogenesis assay based on the use of endothelial cell spheroids, 2010. Simone Kutschera, Functions of Semaphorin 3G during angiogenesis, lymphangiogenesis and tumor development, 2010. Joycelyn Wüstehube, Characterization of cerebral cavernous malformation protein CCM1 in endothelial cells, 2011. Christian Dietz, Analyses of small Rho-GTPases signaling molecules during vertebrate development and angiogenesis, 2011. Arne Bartol, The role of the Angiopoietin-Tie system in blood vessel maturation and maintance, 2012. Anja Weick, Identification of Semaphorin 3G as a novel regulator of tumor lymphangiogenesis and metastasis, 2013. Matthias Wieland, Role of hepatic stellate cell-expressed endosialin in liver health and disease, 2013. Sonija Savant, The role of Tie1 receptor in Angiopoietin-Tie2 signaling during vascular morphogenesis, 2013. Current extramural funding German Research Council: Vascular differentiation and Remodeling; Speaker project within SFB/TR 23 (2009-2017). German Research Council: Role of the Angiopoietin/Tie2 system in controlling vascular morphogenesis and homeostasis; project within the SFB-TR23 (2005-2017). APPENDIX II EU FP7 Project SyStemAge: “Systems Biology of Stem Cell Ageing” (2013-2017). Helmholtz Alliance: Preclinical Comprehensive Cancer Center” (2013-2016). Leducq Foundation Transatlantic Network of Excellence: Lymph vessels in obesity and cardiovascular research (2012-2016). German Research Council: Tumor-specific vascular reprogramming in HCC; project within the SFB-TR77 (2010 - 2014). German Research Council: The role of Angiopoietin/Tie system in regulating the stem cell niche; SFB873 (2010-2014). 74 DR. IRIS AUGUSTIN Div. Signaling and Functional Genomics DKFZ and University Heidelberg Im Neuenheimer Feld 580 D-69120 Heidelberg 06221 421955 (Fax) 06221 421959 (Fax) [email protected] Curriculum vitae since 2008 2004-2008 2002-2004 2001-2002 1998-2000 1999 1995-1998 1991-1995 1989-1991 Senior Scientist, DKFZ & Heidelberg Univ., Heidelberg Maternaty leave (third child) Postdoctoral Fellow, MPI for Immunology, Freiburg Maternaty leave (second child) Postdoctoral Fellow, MPI for experimental Medicine, Göttingen Otto-Hahn-Medaille of MPG PhD, MPI for experimental Medicine, Göttingen Biology, University Göttingen Biology, University Cologne Areas of research expertise since 2008 Wnt signaling in skin development, embryonic stem cells and tumors 2002-2004 Function of Protocaherins in brain architecture 1995-2000 Neurotransmitter release at synapses Augustin, I, Gross J, Baumann D, Korn C, Kerr G, Grigoryan T, Mauch C, Birchmeier W, Boutros M, 2013 Psoriasiform dermatitis-related phenotype caused by loss of epidermal Wnt secretion. J. Exp. Med. 26:1761-77. (IF 13,21) Augustin, I., V. Goidts, A. Bongers, G. Kerr, G. Vollert, B. Radlwimmer, C. Hartmann, C. HeroldMende, G. Reifenberger, A. von Deimling, and M. Boutros. 2012. The Wnt secretion protein Evi/Gpr177 promotes glioma tumourigenesis. EMBO molecular medicine 4:38-51. (IF 7,80) Augustin, I., S. Korte, M. Rickmann, H.A. Kretzschmar, T.C. Sudhof, J.W. Herms, and N. Brose. 2001. The cerebellum-specific Munc13 isoform Munc13-3 regulates cerebellar synaptic transmission and motor learning in mice. The Journal of neuroscience: The official journal of the Society for Neuroscience 21:10-17. (IF 6,91) Augustin, I., C. Rosenmund, T.C. Sudhof, and N. Brose. 1999b. Munc13-1 is essential for fusion competence of glutamatergic synaptic vesicles. Nature 400:457-461. (IF 38.60) 75 APPENDIX II 5 selected (most important) publications Korn C, Scholz B, Hu J, Srivastava K, Wojtarowicz J, Arnsperger T, Adams R, Boutros M, Augustin HG, Augustin I (2014) Endothelial cell derived non-canonical Wnt ligands control vascular pruning in developmental angiogenesis. Development in press (IF 6,9) PhD/MD students (last 5 years) and titles of their theses Claudia Korn (2001-2014) Endothelial cell-derived non-canonical Wnt ligands control vascular pruning in angiogenesis Master students and titles of their theses Dyah Dewi (01/2013 – 09/2013) Analysis of Evi in embryonic stem cells and teratoma Daniel Baumann (02/2012 – 09/2012) Functional analysis of Evi in epithelial knockout mice Gordon Vollert (09/2010 – 06/2011) Functional analysis of Evi in transgenic embryonic stem cells and mice Nesrin Tuysuz (10/2009 – 04/2010) Generation and characterization of Evi-transgenic embryonic stem cells APPENDIX II 76 PROF. DR. RER. NAT. MICHAEL BOUTROS Division Signaling and Functional Genomics German Cancer Research Center (DKFZ) 69120 Heidelberg, Germany Department Cell and Molecular Biology Medical Faculty Mannheim Heidelberg University +49-(0) 6221-42-1951 (Phone) +49-(0) 6221-42 1959 (Fax) [email protected] Curriculum vitae 2008 – to date 2008 – to date 2003 – 08 1999 – 03 1999 – 01 1996 – 99 1995 – 96 1994 – 95 1993 – 96 1991 – 93 Head of Division, German Cancer Research Center (DKFZ) Professor and Chair, Cell and Molecular Biology, University of Heidelberg, Medical Faculty Mannheim Independent Group leader, German Cancer Research Center (DKFZ) Heidelberg, Germany. Postdoctoral fellow, Harvard Medical School, USA M.P.A., Kennedy School of Government, Harvard University, USA Ph.D., European Molecular Biology Laboratory and Heidelberg University, Heidelberg Diploma thesis research, Cold Spring Harbor Laboratory, USA Fulbright Exchange Program, SUNY Stony Brook, USA Studies in Biochemistry, University Witten/Herdecke Studies in Biology, RWTH Aachen 5 selected (most important) publications Augustin, I., Gross, J., Baumann, D., Korn, C., Kerr, G., Grigoryan, T., Mauch, C., Birchmeier, W., and M. Boutros. 2013. Loss of epidermal Evi/Wls results in a phenotype resembling psoriasiform dermatitis. Journal of Experimental Medicine 210:1761-77. (IF 13.21) Gross, J.C., V. Chaudhary, K. Bartscherer, and M. Boutros. 2012. Active Wnt proteins are secreted on exosomes. Nat Cell Biol. 14:1036–1045. (IF 20.76) Bartscherer, K., N. Pelte, D. Ingelfinger, and M. Boutros. 2006. Secretion of Wnt Ligands requires Evi, a conserved transmembrane protein. Cell 125:523-533. (IF 31.96) Boutros, M., A.A. Kiger, S. Armknecht, K. Kerr, M. Hild, B. Koch, S.A. Haas, R. Paro, and N. Perrimon. 2004. Genome-wide RNAi analysis of growth and viability in Drosophila cells. Science 303:832-835. (IF 31.03) Boutros M., N. Paricio, D. Strutt, and M. Mlodzik. 1998. Dishevelled activates JNK and discriminates between JNK pathways in planar polarity and wingless signaling. Cell 94:109-18. (IF 31.96) 77 APPENDIX II Areas of research expertise Wnt signaling in development and cancer, systems genetics and synthetic lethality, high-throughput screening and high-throughput imaging PhD students (last 5 years) and titles of their theses Kubilay Demir, Identification and functional analysis of RAB8B as a regulator of WNT/beta-Catenin signaling pathway, 2007 – 2012, magna cum laude Thomas Horn, Mapping of signaling networks through synthetic genetic interaction analysis by RNAi, 2007 – 2010, summa cum laude Tina Buechling, Characterization of novel mediators of the Wnt/Frizzled signal transduction cascade, 2006 – 2010, summa cum laude Dorothee Nickles, Identification of Novel Regulators of TNF-α Signaling using Genome-wide RNAi Screens, 2006 – 2010, magna cum laude Zeynep Arziman, Systematic analysis of RNAi experiments and deep sequencing data, 2004 – 2008, magna cum laude Sandra Steinbrink, Identification of modifiers of cellular viability and TRAIL-induced apoptosis using genome-wide RNAi screens, 2004- 2008, magna cum laude Kerstin Bartscherer, Identification and characterization of Evi, a novel regulator of Wnt secretion, 2004 – 2007, summa cum laude David Kuttenkeuler, Dissection of Nuclear Factor kB Pathways in Drosophila Innate Immunity, 2003 – 2007, magna cum laude Viola Gesellchen, Identification of new modifiers of the Imd immune signaling pathway, 2003 – 2007, magna cum laude Current extramural funding ERC Advanced Grant, Synthetic Genetic Interaction Analysis, 2012-2017, PI Hartmut-Hoffmann Berling International Graduate School for Molecular and Cellular Biology (HBIGS), Excellence Initiative (DFG/BMBF), Principal Investigator and Member Executive Board (2007-2012) APPENDIX II Excellence Cluster CellNetworks, 2010-2012, DFG, Principal Investigator and Member of the Steering Committee Forschergruppe Wnt FOR1036, Mechanisms, functions and evolution of Wnt signaling pathways, 2012-2014, DFG, BO1791/4-2, PI Collaborative Research Center 873, Maintenance and Differentiation of Stem Cells in Development and Disease, DFG, 2010-2014, PI 78 PD DR. ADELHEID CERWENKA German Cancer Research Center Boveri Junior Research Group/DKFZ/D080 Im Neuenheimer Feld 280 D-69120 Heidelberg Phone: +49 6221 42 4480 Fax: +49 6221 42 3759 [email protected] Curriculum vitae 2007 Venia Legendi for Immunology, University of Heidelberg, Faculty of Medicine, Heidelberg, Germany 2003 - today Head of Boveri Junior Group “Innate Immunity” at the German Cancer Research Center, Heidelberg, German 2001 - 2003 Head of Laboratory, Division of Autoimmune Diseases, Novartis Research Institute, Vienna, Austria 1998 - 2001 Post-doc, DNAX Research Institute and University of California, San Francisco, USA, with Prof. Lewis L. Lanier 1996 - 1998 Post-doc, University of California, San Diego, CA, USA and at the Trudeau Institute, NY, USA, with Prof. Richard W. Dutton 1991 - 1995 PhD, Institute of Immunology, University of Vienna, with Prof. Walter Knapp 1986 - 1991 Diploma in Pharmacy, Institute of Immunology, University of Vienna, Austria 1986 Abitur, Piaristengymnasium Krems, Austria, Golden Ring Award “summa cum laude” 5 selected (most important) publications Fiegler N, Textor S, Arnold A, Rölle A, Oehme I, Breuhahn K, Moldenhauer G, Witzens-Harig M, Cerwenka A. 2013. Downregulation of the activating NKp30 ligand B7-H6 by HDAC inhibitors impairs tumor cell recognition by NK cells, Blood, Aug 1;122(5):684-93. (IF 9.0) J. Ni, M. Miller, A. Stojanovic, N. Garbi, A. Cerwenka. 2012. Sustained effector function of IL12/15/18 preactivated NK cells against established tumors” J Exp Med 209(13):2351-65. (IF 13.2) APPENDIX II Areas of research expertise Tumor immunity, Innate immunity, Natural Killer cells Textor S, Fiegler N, Arnold A, Porgador A, Hofmann TG, Cerwenka A. 2011. Human NK cells are alerted to induction of p53 in cancer cells by up-regulation of the NKG2D-ligands ULBP1 and ULBP2, Cancer Res. 71(18):5998-6009. (IF 8.6) N. Nausch, I. E Galani, E. Schlecker and A. Cerwenka 2008. Mononuclear Myeloid-Derived “Suppressor” Cells express RAE-1 and activate NK cells. Blood 112(10):4080-9. (IF 9.0) A. Cerwenka, A. B. H. Bakker, T. McClanahan, J. Wagner, J. Wu, J. H. Phillips, and L. L. Lanier 2000. Retinoic acid early inducible genes define a ligand family for the activating NKG2D receptor in mice (2000). Immunity. 12: 721-727. (IF 19.8) 79 PhD/MD students (last 5 years) and titles of their theses PhD Anja Tessarz, 2007, Identifying key players in TREM-1/DAP12 signalling Sonja Textor, 2008, Role of NK cells and NK cell receptor ligands in cervical carcinogenesis Kai Zanzinger, 2008, Expression and signalling of triggering receptor expressed on myeloid cells (TREM)-1 Norman Nausch, 2008, The importance of NKG2D-RAE-1 in anti-tumor immune response Ioanna Evdokia Galani, 2008, Enhancing anti-tumor immunity to MHC class-I-deficient tumors: role of regulatory T cells and type I IFN Marco Wendel, 2009, Mechanism of natural killer cell accumulation in tumors Ana Stojanovic, 2009, Molecular signature of tumor-infiltrating NK cells Eva Schlecker, 2011, The role of tumor-infiltrating MDSC subsets in tumor progression Nathalie Fiegler, 2013, Expression, regulation, and function of the Natural Killer Cell ligand B7-H6 in tumor cells Current extramural funding German Carreras Foundation, Deutsche Carreras Stiftung Leukämie-Stiftung e.V: DJCLS R 11/06: Activation of Natural Killer cells by NKp30/B7-H6 in haematological neoplasia, 2011-2014 Joint Project: DKFZ-Bayer Health Care, 2011-2014 APPENDIX II Joint Project: DKFZ-Karolinska Institute, Role of Natural Killer cells in hepatocellular cancer (HCC) pathogenesis, 2012-2014 German Cancer Aid, Deutsche Krebshilfe, 110442, Generation of human CD8+ T lymphocytes co-expressing antigen-specific T cell receptors and activating NK cell receptors for immunotherapy of cancer, 2013-2016 80 PROF. DR. MED. ALEXANDER H. ENK Curriculum vitae 2004-to date 1997 1995 1993 1993 1992-1994 1990-1992 1988-1990 1988 1988 1984-1988 1983 – 1988 1982 – 1988 Appointment as Chairman and Full Professor, Dept. of Dermatology and Venerology, Ruprecht-Karls-University of Heidelberg Associate Professor and Vice Chairman, Dept. of Dermatol, JohannesGutenberg University of Mainz Venia legendi for Dermatology and Venerology, Johannes-Gutenberg University of Mainz, “Frühe molekulare Veränderungen in der Induktionsphase der allergischen Kontaktdermatitis”1994-1997 Assistant Professor, Dept. of Dermatol., Johannes-Gutenberg University of Mainz Board Certification Allergology Board Certification Dermatology and Venerology Resident Dept. of Dermatol., Johannes-Gutenberg the University of Mainz1992 Forgarty Scholarship of the National Institutes of Health (NIH) Postdoctoral Fellowship, Dermatol. Branch of the National Institutes of Health (NIH), DFG scholarship “Arzt im Praktikum”, Dept. of Dermatol. Johannes-Gutenberg University of Mainz Fullbright Scholarship “Arzt im Praktikum”, Dept. of Dermatol., University Hospital Münster Promotion to Dr. med. , Dept. of Dermatol. of the University of Münster), Thesis: “Produktion von IFN-γ durch epidermale LC nach Stimulation” Scholarship of the Studienstiftung des Deutschen Volkes (German Research Foundation) Medical School, Westf. Wilhelms-Universität Münster Areas of research expertise Dermato-Oncology, Contact Hypersensitivity, Inflammation, Immunology 5 selected (most important) publications Mahnke K., Qian Y., Fondel S., Brueck J., Becker C. and A.H. Enk. 2005. Targeting of antigens to activated dendritic cells in vivo cures metastatic melanoma in mice. Cancer Res. 1;65(15): 700712. (IF 8,65) Mahnke K., Qian Y. Knop J., and A.H. Enk .2003. Dendritic cells, engineered to secrete a T-cell receptor mimic peptide, induce antigen-specific immunosuppression in vivo. Nat Biotechnol. 21(8):903-8. (IF 32,43) Mahnke K., Qian Y., Knop J. and A.H. Enk. 2003. Induction of CD4+/CD25+ regulatory T cells by targeting of antigens to immature dendritic cells. Blood 15;101(12):4862-9. (IF 9,06) 81 APPENDIX II Chairman and Full Professor Department of Dermatology University Medical Center Heidelberg, Ruprecht-Karls-University of Heidelberg Voßstrasse 2 69115 Heidelberg 06221 56 8501 (Fon) 06221 56 5406 (Fax) [email protected] Jonuleit H., Schmitt E., Kakirman H., Stassen M., Knop J. and A.H. Enk. 2002.Infectious tolerance: human CD25(+) regulatory T cells convey suppressor activity to conventional CD4(+) T helper cells. J Exp Med.15;196(2):255-60. (IF 13,21) Jonuleit H., Schmitt E., Stassen M., Tuettenberg A., Knop J. and A.H. Enk. 2001. Identification and functional characterization of human CD4(+)CD25(+) T cells with regulatory properties isolated from peripheral blood. J Exp Med. 4;193(11):1285-94. (IF 13,21) PhD/MD students (last 5 years) Michael Maas, Ph.D., 2010 Volker Storn, Ph.D, 2008 . Sonja Schallenberg, Ph.D.,2008 Kurt Schönfeld, Ph.D., 2008 Yingjie Qian, PhD., 2005 Current extramural funding Helmholtz Alliance (HA-202), Melanoma Therapy Targeting, Förderperiode bis 31.12.12 SFB 938 Milieuspezifische Kontrolle immunologischer Reaktivität, 1. Förderperiode bis 31.12.2014 APPENDIX II 82 DR. MED. MORITZ FELCHT Resident of Dermatology Department of Dermatology, Venereology, and Allergy, University Medical Center Mannheim, Heidelberg University Theodor-Kutzer-Ufer 1-3 68135 Mannheim 0621 383 2280 (Fon) 0621 383 3815 (Fax) [email protected] Curriculum Vitae 2008 – to date 2007 Education 2002-2006 2000-2002 02-04/2006 12-01/2005/06 02-03/2004 08-09/2001 Board-certified dermatologist Resident of Dermatology Department of Dermatology, Venerology, Allergy University Medical Center Mannheim, Heidelberg University (Prof. Goerdt) Postdoctoral Studies at Vascular Biology & Tumor Angiogenesis Medical Faculty Mannheim, Heidelberg University (CBTM) & German Cancer Research Center, Heidelberg (Prof. Augustin) Resident of General Surgery University of Schleswig-Holstein, Campus Lübeck (Prof. Bruch) Julius-Maximilians-University, Würzburg Freie University Berlin Strong Memorial Hospital, University of Rochester, USA Boston University Medical School, USA St. Vincent’s Hospital, Geelong, University of Melbourne (General Medicine), Australia Bristol Royal Infirmary Hospital (Admission Ward), England Areas of research expertise Cancer biology: Vascular Tumor Microenvironment: since 06/2013 Group Leader of the Juniorgroup “Vascular Tumor Microenvironment” at the Department of Dermatology/ Vascular Biology & Tumor Angiogenesis Medical Faculty Mannheim, Heidelberg University (CBTM); funded by the DFG; Tumor angiogenesis: 2008-06/2013 Post-Doctoral studies at the Department of Vascular Biology & Tumor Angiogenesis Medical Faculty Mannheim, Heidelberg University (CBTM) & German Cancer Research Center, Heidelberg (Prof. Augustin); Cutaneous Tumor Development: since 2007 Member of the Working Group of Cutaneous Lymphomas, University Medical Center Mannheim (Prof. Klemke and Prof. Goerdt); Apoptosis and Cell Death: 20022007 The medical thesis (Dr. med.) analysed “death-receptor-mediated MAP-kinase-activation in keratinocytes” („Todesrezeptor-vermittelte MAP-Kinasen-Aktivierung in Keratinozyten“; Department of Dermatology, Venerology, Allergy University of Würzburg (Prof. Leverkus) 83 APPENDIX II Professional Experience since 12/2012 2007- to date 5 selected (most important) publications Felcht, M.*, W. Koenen*, C. Weiss, K. Weina, C. Geraud, and J. Faulhaber. (2013) Delayed closure of complex defects with serial tightening of loop sutures – clinical outcome in 64 consecutive patients. J Eur Acad Dermatol doi: 10.1111/jdv.12122; 2013 (IF 2.70) *equal contribution. Felcht, M., R. Luck, A. Schering, P. Seidel, K. Srivastava, J. Hu, A. Bartol, Y. Kienast, C. Vettel, E.K. Loos, S. Kutschera, S. Bartels, S. Appak, E. Besemfelder, D. Terhardt, E. Chavakis, T. Wieland, C. Klein, M. Thomas, A. Uemura, S. Goerdt, and H.G. Augustin. (2012) Angiopoietin-2 differentially regulates angiogenesis through TIE2 and integrin signaling. J Clin Invest. 122:19912005 (IF 12, 81). Felcht,* M., M. Heck*, C. Weiss, J. C. Becker, E. Dippel, C. S. L. Müller, D. Nashan, M. M. Sachse, J. P. Nicolay, N. Booken, S. Goerdt, and C.-D. Klemke. 2012 Expression of the T-cell regulatory marker FOXP3 in primary cutaneous large B-cell lymphoma cells. Brit J Dermatol. 167: 348-59 (IF 3,76). *equal contribution Felcht M., N. Booken, P. Stroebel, S. Goerdt, and C.D. Klemke. 2011 The value of molecular diagnostics in primary cutaneous B-cell lymphoma in the context of clinical findings, histology, and immunohistochemistry. J Am Acad Dermatol 64: 135-43. (IF 4,91) Thomas M.*, M. Felcht*, K. Kruse, S. Kretschmer, C. Deppermann, A.V. Benest, U. Fiedler, and H.G. Augustin. 2010 Angiopoietin-2 stimulation of endothelial cells induces alphavbeta3 integrin internalization and degradation. J Biol Chem 285: 23842-9. *equal contribution (IF 4,65) PhD/ MD students Martin Petkov: PhD: “Analyse des Einflusses der Angiopoietin-like Proteine-3 und -4 auf die Angiopoietin-2 Signalgebung”, since 2014 Malte Kranert: MD: “Analyse des Tumor-assozierten Gefäßnetzes beim kutanen T-Zell Lymphom”, since 2013 Christine Stumpf: MD: “Analyse des Tumor-assozierten Gefäßnetzes beim kutanen B-Zell Lymphom”, since 2011 (together with Prof. Klemke) APPENDIX II Robert Luck: Bachelor thesis: “Effects of Angptl-3 and Angptl-4 stimulation of brain pericytes”, 0209/2011 (together with Prof. Augustin) Philipp Seidel: Master thesis: “Tie2 independent functions of angiopoietin-2 during angiogenesis”, 04-10/2010 (together with Prof. Augustin) Current extramural funding “Analyse des Einflusses der Angiopoietin-like Proteine-3 und -4 auf die Angiopoietin-2 Signalgebung”, 2013-2016 DFG Erstantrag FE 1282/1-1 84 PD DR. MED. CYRILL GÉRAUD Consultant Physician Department of Dermatology, Venereology, and Allergology, University Medical Center Mannheim, Heidelberg University Theodor-Kutzer-Ufer 1-3 68135 Mannheim 0621 383 2126 (Fon) 0621 383 3815 (Fax) [email protected] Habilitation (Venia legendi) for Dermatology and Venereology (Medical Faculty Mannheim, Heidelberg University) 05/2013 - to date Consultant physician (Oberarzt), Department of Dermatology, Venereology, and Allergology, University Medical Center Mannheim, Heidelberg University 04/2013 Board Certification in Dermatology and Venereology 09/2012- 04/2013 Senior Resident physician (Funktionsoberarzt), Department of Dermatology, Venereology, and Allergology, University Medical Center Mannheim, Heidelberg University 2008 - 2013 Resident physician, Department of Dermatology, Venereology, and Allergology, University Medical Center Mannheim, Heidelberg University 2009 - to date Group leader research group “Organ-specific microvessels” with Prof. Dr. Sergij Goerdt, Department of Dermatology, Venereology, and Allergology, University Medical Center Mannheim, Heidelberg University 2009 Doctoral thesis: Institute of Anatomy und Cell Biology, Department of Neuroanatomy (Prof. M. Frotscher) of the Albert-Ludwigs-University Freiburg, Germany: “Involvement of the extracellular matrixprotein Reelin during differentiation of hippocampal neurons in vitro” (magna cum laude) 2001-2007 Medical Degree: Albert-Ludwigs University Medical School in Freiburg im Breisgau, Germany with extramural electives in San Diego, CA, USA Internal Medicine) and Tampa, FL, USA (Dermatology) Areas of research expertise Endothelial cells, Organ-Specific Endothelial Differentiation and Function, Dermato-Oncology, Dermatopathology, Metastasis. 5 selected (most important) publications Géraud, C.*, K. Schledzewski*, A. Demory, D. Klein, M. Kaus, F. Peyre, C. Sticht, K. Evdokimov, S. Lu, A. Schmieder, and S. Goerdt. 2010. Liver sinusoidal endothelium: a microenvironmentdependent differentiation program in rat including the novel junctional protein liver endothelial differentiation-associated protein-1. Hepatology. 52:313-26. (IF 12.00) Schledzewski, K.*, C. Géraud*, B. Arnold, S. Wang, H.J. Grone, T. Kempf, K.C. Wollert, B.K. Straub, P. Schirmacher, A. Demory, H. Schonhaber, A. Gratchev, L. Dietz, H.J. Thierse, J. Kzhyshkowska, and S. Goerdt. 2011. Deficiency of liver sinusoidal scavenger receptors stabilin-1 and -2 in mice causes glomerulofibrotic nephropathy via impaired hepatic clearance of noxious blood factors. J Clin Invest. 121:703-14. (* K.S. and C.G. contributed equally to this work) (IF 12.812) 85 APPENDIX II Curriculum vitae 02/2014 Géraud, C.*, C. Mogler*, A. Runge, K. Evdokimov, S. Lu, K. Schledzewski, B. Arnold, G. Hämmerling, P.S. Koch, K. Breuhahn, T. Longerich, A. Marx, C. Weiss, F. Damm, A. Schmieder, P. Schirmacher, H.G. Augustin, S. Goerdt. 2013. Endothelial transdifferentiation in hepatocellular carcinoma: loss of Stabilin-2 expression in peri-tumourous liver correlates with increased survival. Liver Int. 33:1428-40. (IF 3.870) Géraud, C.*, K. Evdokimov*, B.K. Straub, W.K. Peitsch, A. Demory, Y. Dorflinger, K. Schledzewski, A. Schmieder, P. Schemmer, H.G. Augustin, P. Schirmacher, and S. Goerdt. 2012. Unique cell type-specific junctional complexes in vascular endothelium of human and rat liver sinusoids. PLoS One. 7:e34206. (IF 3.730) Schmieder, A., K. Schledzewski, J. Michel, J.P. Tuckermann, L. Tome, C. Sticht, C. Gkaniatsou, J.P. Nicolay, A. Demory, J. Faulhaber, J. Kzhyshkowska, C. Géraud, and S. Goerdt. 2010. Synergistic activation by p38MAPK and glucocorticoid signaling mediates induction of M2-like tumor-associated macrophages expressing the novel CD20 homolog MS4A8A. Int J Cancer. (IF 6.198) PhD/MD students (last 5 years) and titles of their theses PhD Siladitta Biswas; Functional characterization of the novel junctional protein liver endothelial differentiation-associated protein (leda)-1, 2011-2014, together with Prof. Dr. S. Goerdt Francis Peyre, Tumor-induced endothelial differentiation in vitro, 2007 - 2011, cum laude, together with Prof. Dr. S. Goerdt Johanna Zierow,; Molecular mechanisms of liver sinusoidal endothelial differentiation in development, 2013-2017, together with Prof. Dr. S. Goerdt MD APPENDIX II Claudia Ansorge, Organ-spezifische Endothelzelldifferenzierung in vitro, 2010 - 2014, together with Prof. Dr. S. Goerdt Konstantin Evdokimov, Characterization of a novel liver endothelial differentiation-associated protein1, 2009 – 2013, summa cum laude, together with Prof. Dr. S. Goerdt Shun Lu, Characterization of tumor endothelium in a murine model of hepatocellular carcinoma and in hepatic melanoma metastasis, 2009 – 2010, rite, together with Prof. Dr. S. Goerdt Manuel Winkler, Functional analysis of liver endothelial differentiation associated protein (Leda) -1 in malignant melanoma, 2010 - 2014, together with Prof. Dr. S. Goerdt Current extramural funding Molekulare und funktionelle Analyse des neuen junktionalen Proteins Leda-1 in Endothelzellen und beim malignen Melanom, 2011 – 2014, DFG-Erstantrag GE-2339/1-1 Liver-specific vascular differentiation and function, 2013-2017, Project B1, Collaborative Research Center (DFG Sonderforschungsbereich) / Transregio 23 (together with Prof. Dr. S. Goerdt) 86 DR. PETER GESERICK Section of Molecular Dermatology, Department of Dermatology, Venereology, and Allergy, University Medical Center Mannheim, Heidelberg University Theodor-Kutzer-Ufer 1-3 68135 Mannheim 0621 383 3990 [email protected] Curriculum vitae 01.2010 - to date 01.2012 - 06.2012 01.2006 - 12.2009 01.2005 - 12.2005 01.2001 - 12.2004 10.1996 - 09.2000 Postdoctoral Studies, Section of Molecular Dermatology, Department of Dermatology, Venereology, and Allergy, University Medical Center Mannheim, Heidelberg University (Mannheim) Parental leave Postdoctoral Studies, Otto-von-Guericke-University (Magdeburg) Postdoctoral Studies, Max-Planck-Institute for Infection biology (Berlin) PhD in biochemistry to Dr. rer. nat, Max-Planck-Institute for Infection biology and Free University of Berlin (Berlin) Study of Biochemistry, University of Potsdam Areas of research expertise Signal transduction, Cell death, Inflammation, tumorigenesis, SCC, melanoma Feoktistova M.*, P.Geserick*, B.Kellert, D.Panayotova-Dimitrova, C.Langlais, M.Hupe, K.Cain, M.MacFarlain, G.Häcker, and M.Leverkus. 2011. cIAPs block Ripoptosome formation, a RIP1/caspase 8 containing intracellular cell death complex differentially regulated by cFLIP isoforms. Mol. Cell. 43(3):449-63. (* equal contribution) (IF 15,28) Geserick, P., M.Hupe, M.Moulin, and M.Leverkus. 2010. RIP-in CD95L-induced cell death: The control of alternative death receptors pathways by cIAPs. Cell Cycle. 9(14):2689-2691. (IF 5,24) Geserick, P., M.Hupe, M.Moulin, W.W.Wong, M.Feoktistova, B.Kellert, H.Gollnick, J.Silke, and M.Leverkus. 2009. Cellular IAPs inhibit a cryptic CD95-induced cell death by limiting RIP1 kinase recruitment. J Cell Biol. 187:1037-1054. (IF 10,82) Geserick, P., C.Drewniok, M.Hupe, T.L.Haas, P.Diessenbacher, M.R.Sprick, M.P.Schon, F.Henkler, H.Gollnick, H.Walczak, and M.Leverkus. 2008. Suppression of cFLIP is sufficient to sensitize human melanoma cells to TRAIL or CD95L-mediated apoptosis. Oncogene. 27:32113220. (IF 7,36) 87 APPENDIX II 5 selected (most important) publications Feoktistova M.*, P.Geserick*, D.Panayotova-Dimitrova, and M.Leverkus. 2012. Pick your poison: the Ripoptosome, a cell death platform regulating apoptosis and necrosis. Cell Cycle. 11(3):460-7. (* equal contribution) (IF 5,24) PhD (last 5 years) and titles of their theses PhD Jing Wang, Role of BRAF and MEK kinases for death receptor induced cell death in melanoma. (in progress) Ramon Schilling, Identification and characterization of novel molecules involved necroptotic signalling patways in keratinocytes and malignant skin cancer cells, 2012-2015 (in progress) Sebastian Horn, Identification and characterization of caspase 10 isoforms for cell death signalling patways in keratinocytes and skin malignancies, 2011-2014 (in progress) Mike Hupe, The role of DISC-associated signals for apoptotic and non-apoptotic signalling in keratinocytes, 2006 - 2009, summa cum laude Maria Feoktistova, The contribution of RIP1 for TRAIL- and TLR3-mediated death signalling in human keratinocytes, 2005 -2010, magna cum laude Shyam Kavuri, cFLIP and its splice variants and their role for death receptor-mediated NF-κB and JNK activation in the skin, 2005 - 2009, cum laude APPENDIX II 88 PROF. DR. MED. SERGIJ GOERDT Chairman, Dept. of Dermatology, Venereology, and Allergy, University Medical Center Mannheim, Heidelberg University Theodor-Kutzer-Ufer 1-3 68135 Mannheim 0621 383 2280 (Fon) 0621 383 3815 (Fax) [email protected] Curriculum vitae since 2013 since 2009 2007-2012 2006-2010 2005-2009 2002-2009 seit 2000 1997-1999 1997 1995-2000 1989-1991 1987/89, 1991/94 1985-1987 1978-1985 Vice Dean, Medical Faculty Mannheim, Heidelberg University Member of the Board of Collaborative Research Center TRR 23 Speaker of the Excellence Center Dermatology Mannheim of the State of Baden-Württemberg Vice Dean for Structure and Development, Med. Faculty Mannheim Vice Speaker Collaborative Research Center TRR 23 “Vascular Differentiation and Remodelling” Member of the Board of Collaborative Research Center SFB 405 “Immuntoleranz und ihre Störungen” Chairman, Dept. Dermatology, University Medical Center Mannheim, Heidelberg University President, Dermatological Society of Berlin Associate Professor for Dermatology and Venereology, FU Berlin Vice Chairman, Dept. Dermatology, UKBF, FU Berlin Visiting Research Fellow, Harvard Medical School, Boston, MA Resident, Dept. Dermatology, University of Münster Research Assistent, Inst. f. Exp. Dermatology, University of Münster Medical College, Universities of Münster, Mainz, Wien, and Freiburg 5 selected (most important) publications Schledzewski, K.*, C. Géraud*, B. Arnold, S. Wang, H.J. Grone, T. Kempf, K.C. Wollert, B.K. Straub, P. Schirmacher, A. Demory, H. Schonhaber, A. Gratchev, L. Dietz, H.J. Thierse, J. Kzhyshkowska, and S. Goerdt. 2011. Deficiency of liver sinusoidal scavenger receptors stabilin-1 and -2 in mice causes glomerulofibrotic nephropathy via impaired hepatic clearance of noxious blood factors. J Clin Invest. 121:703-14. (IF 12.81) Géraud, C.*, K. Schledzewski*, A. Demory, D. Klein, M. Kaus, F. Peyre, C. Sticht, K. Evdokimov, S. Lu, A. Schmieder, and S. Goerdt. 2010. Liver sinusoidal endothelium: a microenvironmentdependent differentiation program in rat including the novel junctional protein liver endothelial differentiation-associated protein-1. Hepatology. 52:313-26. (IF 12.00) Klein, D., A. Demory, F. Peyre, J. Kroll, H.G. Augustin, W. Helfrich, J. Kzhyshkowska, K. Schledzewski, B. Arnold, and S. Goerdt. 2008. Wnt2 acts as a cell type-specific, autocrine growth factor in rat hepatic sinusoidal endothelial cells cross-stimulating the VEGF pathway. Hepatology. 47:1018-31. (IF 12.00) Booken, N., A. Gratchev, J. Utikal, C. Weiss, X. Yu, M. Qadoumi, M. Schmuth, N. Sepp, D. Nashan, K. Rass, T. Tüting, C. Assaf, E. Dippel, R. Stadler, CD Klemke, and S. Goerdt. 2008. Sézary syndrome is a unique cutaneous T-cell lymphoma as identified by an expanded gene signature including diagnostic marker molecules CDO1 and DNM3. Leukemia. 22: 393-399. (IF 10.16) 89 APPENDIX II Areas of research expertise Dermato-Oncology, Vascular Biology, Immunology / Innate Immunity Kzhyshkowska, J., S. Mamidi, A. Gratchev, E. Kremmer, C. Schmuttermaier, L. Krusell, G. Haus, J. Utikal, K. Schledzewski, J. Scholtze, S. Goerdt. 2006. Novel stabilin-1 interacting chitinase-like protein (SI-CLP) is upregulated in alternatively activated macrophages and secreted via the lysosomal pathway. Blood. 107:3221-28. (IF 9.06) PhD/MD students (last 5 years) and titles of their theses PhD Biswas, Siladitta; Functional characterization of the novel junctional protein liver endothelial differentiation-associated protein (leda)-1, 2011-2014, together with PD Dr. C. Géraud Dollt, Claudia; Identification and characterization of progression-associated TAM molecules in malignant melanoma, 2013-2017, together with Dr. A. Schmieder Michel, Julia; Characterization of a novel melanoma-associated TAM molecule, 2010-2014, magna cum laude, together with Dr. A. Schmieder Nurgazieva, Dinara; TGFbeta signaling in alternatively activated macrophages, 2008 – 2011, magna cum laude, together with Prof. Dr. J. Kzhyshkowska Peyre, Francis; Tumor-induced endothelial differentiation in vitro, 2007 – 2011, cum laude, together with PD Dr. C. Géraud Popova, Anna; New mechanism of signal transduction in alternatively activated macrophages, 2006 – 2009, magna cum laude, together with Prof. Dr. J. Kzhyshkowska Riabov, Vladimir; Analysis of the role of stabilin-1 in tumour growth and its functions in tumourassociated macrophages, 2008–2011, magna cum laude, together with Prof. Dr. J. Kzhyshkowska Schönhaar, Kathrin; Molecular, phenotypic and functional characterization of the hyaluronan receptor Lyve-1 as a novel TAM molecule in malignant melanoma, 2011-2014, together with Dr. A. Schmieder APPENDIX II Zierow, Johanna; Molecular mechanisms of liver sinusoidal endothelial differentiation in development, 2013-2017, together with PD Dr. C. Géraud Zhang, JingJing: Mechanism of stabilin-1 mediated endocytosis and ligand trafficking, 2006 – 2011, summa cum laude, together with Prof. Dr. J. Kzhyshkowska MD Claudia Ansorge, Organ-spezifische Endothelzelldifferenzierung in vitro, 2010 - 2014, together with PD Dr. C. Géraud Konstantin Evdokimov, Characterization of a novel liver endothelial differentiation-associated protein1, 2009 – 2013, summa cum laude, together with PD Dr. C. Géraud Gkaniatsou, Cleopatra; Ms4a8a overexpression inhibits tumor growth of CT26 colon carcinoma cells in vivo, 2009 – 2010, cum laude, together with Dr. A. Schmieder Kneifel, Simone Alexandra; Standardisierte Versorgung von Weichteildefekten am Schädel, 2007 – 2010, cum laude, together with PD Dr. W. Koenen 90 Linder, Anna Spophie Maria; Retrospektive Analyse von Patienten mit malignen epithelialen Hauttumoren nach Anwendung von plastischen Rekonstruktionsverfahren, 2007 – 2009, magna cum laude, together with PD Dr. W. Koenen Shun Lu, Characterization of tumor endothelium in a murine model of hepatocellular carcinoma and in hepatic melanoma metastasis, 2009 – 2010, rite, together with PD Dr. C. Géraud Manousaridis, Ioannis; Gene expression profiling of alternatively activated human blood monocytes with emphasis on the expression of Wnt-related molecules and FOXQ1. An ex vivo application on acute atopic dermatitis, 2007 – 2009, magna cum laude, together with PD Dr. A. Gratchev Teerling, Gloria-Viktoria; Versorgung schichtübergreifender Weichteildefekte am Schädel mit azellulärem Dermisersatz, 2008 – 2011, cum laude, together with PD Dr. W. Koenen Wen, Ming; Cloning and characterization of inhibitor of DNA binding 3 (ID3) promoter, 2007 – 2010, cum laude, together with PD Dr. A. Gratchev Winkler, Manuel; Functional analysis of liver endothelial differentiation associated protein (Leda) -1 in malignant melanoma, 2010 - 2014, together with PD Dr. C. Géraud Current extramural funding Tumor-associated macrophages and tumor progression: functional plasticity and progressiondependent TAM target molecules in malignant melanoma, 2011-2014, Project H, Collaborative Research Center SFB 938 Tumor-specific vascular reprogramming in HCC: mechanisms and therapeutic targets, 2010-2014, Project C3, Collaborative Research Center TRR77 (together with Prof. Dr. H. Augustin) APPENDIX II Liver-specific vascular differentiation and function, 2013-2017, Project B1, Collaborative Research Center TRR23 (together with PD Dr. C. Géraud) 91 PROF. DR. MED. MARTIN LEVERKUS Associate Professor for Clinical and Molecular Dermatology Section of Molecular Dermatology, Department of Dermatology, Venereology, and Allergy, University Medical Center Mannheim, Heidelberg University Theodor-Kutzer-Ufer 1-3 68135 Mannheim 0621 383 2344 (Fon) 0621 383 4085 (Fax) [email protected] Curriculum vitae 2009 - to date 2006 2004 - 09 2002 2002 2000 2000 1997 1995 - 97 1993 - 94 1993 1985-92 Associate professor (W3) for Clinical and Molecular Dermatology, Medical Faculty Mannheim of the Ruprecht-Karls-University Heidelberg Board approval „Medical Tumor Therapy“ Associate professor (C3) for Clinical and Experimental Dermatology, Otto-von-Guericke-University Magdeburg Venia legendi for Dermatology, University of Würzburg Consultant in Dermatology, Department of Dermatology, University of Würzburg Board certification Allergology Board certification Dermatology Resident in Dermatology, Department of Dermatology, University of Würzburg; Establishment of the independent research group “Apoptosis regulation in the skin” Postdoctoral Studies, Dermatology Department of Boston University, Boston, MA, USA „Arzt im Praktikum“ and Resident in Dermatology, Department of Dermatology, University of Würzburg Promotion to Dr. med. (Physiological Institute of the University of Cologne) MD studies in Cologne, Clermont-Ferrand (France), Johannesburg (South Africa) and Greenville (USA). APPENDIX II Areas of research expertise Dermato-Oncology, Signal tranduction, Cell death, Inflammation 5 selected (most important) publications Feoktistova, M., P.Geserick, B.Kellert, D.P.Dimitrova, C.Langlais, M.Hupe, K.Cain, M.Macfarlane, G.Hacker, and M.Leverkus. 2011. cIAPs block Ripoptosome formation, a RIP1/caspase-8 containing intracellular cell death complex differentially regulated by cFLIP isoforms. Mol. Cell 43:449-463. (IF 15,28) Geserick, P., C.Drewniok, M.Hupe, T.L.Haas, P.Diessenbacher, M.R.Sprick, M.P.Schon, F.Henkler, H.Gollnick, H.Walczak, and M.Leverkus. 2008. Suppression of cFLIP is sufficient to sensitize human melanoma cells to TRAIL or CD95L-mediated apoptosis. Oncogene 27:32113220. (IF 7,36) Geserick, P., M.Hupe, M.Moulin, W.W.Wong, M.Feoktistova, B.Kellert, H.Gollnick, J.Silke, and M.Leverkus. 2009. Cellular IAPs inhibit a cryptic CD95-induced cell death by limiting RIP1 kinase recruitment. J Cell Biol 187:1037-1054. (IF 10,82) Leverkus, M., M.Neumann, T.Mengling, C.T.Rauch, E.B.Bröcker, P.H.Krammer, and H.Walczak. 2000a. Regulation of tumor necrosis factor-related apoptosis-inducing ligand sensitivity in primary and transformed human keratinocytes. Cancer Res. 60:553-559. (IF 8,65) 92 Leverkus, M., H.Walczak, A.McLellan, H.W.Fries, G.Terbeck, E.B.Brocker, and E.Kämpgen. 2000b. Maturation of dendritic cells leads to up-regulation of cellular FLICE- inhibitory protein and concomitant down-regulation of death ligand- mediated apoptosis. Blood 96:2628-2631. (IF 9,06) PhD/MD students (last 5 years) and titles of their theses PhD Mike Hupe, The role of DISC-associated signals for apoptotic and non-apoptotic signalling in keratinocytes, 2006 - 2009, summa cum laude Maria Feoktistova, The contribution of RIP1 for TRAIL- and TLR3-mediated death signalling in human keratinocytes, 2005 -2010, magna cum laude Shyam Kavuri, cFLIP and its splice variants and their role for death receptor-mediated NF-κB and JNK activation in the skin, 2005 - 2009, cum laude Beate Kellert, Die Rolle des Inflammasoms bei Differenzierung, Reifung und Funktion von Dendritischen Zellen, 2007 - 2011 MD Michael Czisch, Die Rolle von Bcl-2 bei der UV- und TRAIL-induzierten Apoptose, 2000 - 2002, magna cum laude Moritz Felcht, Die Todesrezeptor-vermittelte MAPK-Aktivierung in humanen Keratinozyten, 2002 2004, magna cum laude Dominikus Hausmann, Der Einfluss von cFLIP bei der apoptotischen und nichtapoptotischen Signalgebung durch TRAIL-Rezeptoren in Keratinozyten, magna cum laude Current extramural funding Die Bedeutung von A20 und ABIN-1 für die TNF-vermittelte apoptotische und inflammatorische Signalgebung von Keratinozyten, 2011 – 2014, DFG Projekt Le 953/6-1. Generierung und Analyse konditional induzierbarer transgener Tiere zur Funktionsanalyse von cFLIP in der Haut, 2013 – 2016, DFG Projekt Le 953/8-1. Resistenz gegenüber Tumor-spezifischen chimären T-Zellen: synergistische Apoptose-Induktion durch gezielte Hemmung anti-apoptotischer Proteine, 2012 – 2015 (Dt. Krebshilfe, Projekt 109891; gemeinsam mit Prof. Marx, Institut für Pathologie) 93 APPENDIX II Barbara Kehler, Zur Bedeutung des TRAIL-R1 bei der Pathogenese und Tumorprogression des malignen Melanoms, 2007 - 2010 DR. MED. ANKE S. LONSDORF Attending Physician and Junior Group Leader Department of Dermatology University Medical Center Heidelberg, Ruprecht-Karls-University of Heidelberg Voßstrasse 2 69115 Heidelberg 06221 56 8505 (Fon) 06221 56 8083 (Fax) [email protected] Curriculum vitae 10/2011-date 12/2011- 12/2013 12/2011 02/2011 2008 – 02/11 02/06 – 09/10 2006 – 2008 2006 2004 – 2006 2000 – 2001 04/98-06/04 APPENDIX II 1997-2004 Attending Physician, Dept..of Dermatology, Ruprecht-Karls-University of Heidelberg and National Centre for Tumor Diseases Heidelberg (NCT), Germany Olympia Morata Scholarship of Ruprecht-Karls-University of Heidelberg Board Certification Medical Tumor Therapy Board Certification Dermatology and Venerology Residency, Dept. of Dermatology, Ruprecht-Karls-University of Heidelberg, Germany NIH/DFG Research Career Transition Award of the National Institutes of Health (NIH) and the German Research Foundation (DFG) Postdoctoral research fellowship, National Institutes of Health (NIH), National Cancer Institute (NCI), Department of Dermatology, Bethesda, MD, USA Promotion to Dr. med. (Dept. of Internal Medicine, University of Ulm, Germany), Thesis: “Intratumor CpG-oligodeoxynucleotide injection induces protective antitumor T cell immunity” Residency, Dept. of Dermatology, Ruprecht-Karls-University of Heidelberg, Germany Doctoral research fellowship, Case Western Reserve University (CWRU), Dept. of Pathology, Cleveland, OH, USA Scholarship of the German National Academic Foundation (Studienstiftung des deutschen Volkes) Medical School, Ruprecht-Karls-University of Heidelberg, Germany Areas of research expertise Dermato-Oncology, Chemokine Biology, Inflammation, Immunology 5 selected (most important) publications Lonsdorf AS, Kraemer BF, Fahrleitner M, Schoenberger T, Gnerlich S, Ring S, Gehring S, Schneider SW, Kruhlak MJ, Meuth SG, Nieswandt B, Gawaz M, Enk AH, Langer HF: Engagement of αIIbβ3 (GPIIb/IIIa) with ανβ3 mediates interaction of melanoma cells with platelets - a connection to hematogeneous metastasis. J Biol Chem. 2012 Jan 13;287(3):2168-78. (IF 4,65) Chien A.J.*, Moore E.C.*, Lonsdorf A.S., Kulikauskas R.M., Rothberg B.G., Berger A.J., Major M.B., Hwang S.T., Rimm D.L. and R.T. Moon. 2009. Activated Wnt/beta-catenin signaling in melanoma is associated with decreased proliferation in patient tumors and a murine melanoma model. Proc Natl Acad Sci U S A. 27;106(4):1193-8. (IF 9.74) *equal contribution, alphabetical order Hedrick, M.N.*, Lonsdorf A.S. *, Shirakawa A.K., Lee Richard C.C, Liao F., Singh S.P., Zhang H.H., Grinberg A., Love P.E., Hwang S.T. and JM Farber. 2009. CCR6 is required for IL-23- 94 induced psoriasis-like inflammation in mice. J Clin Invest 119:2317-2329. (IF 12,81) * equal contribution, alphabetical order Huang, V., Lonsdorf A.S., L. Fang, T. Kakinuma, V.C. Lee, E. Cha, H. Zhang, K. Nagao, M. Zaleska, W.L. Olszewski, and S.T. Hwang. 2008. Cutting edge: rapid accumulation of epidermal CCL27 in skin-draining lymph nodes following topical application of a contact sensitizer recruits CCR10-expressing T cells. J Immunol 180:6462-6466. (IF 5,52) Lonsdorf, A.S., H. Kuekrek, Stern B.V., Boehm B.O,. Lehmann P.V and M. Tary Lehmann. 2003. Intratumor CpG-oligodeoxynucleotide injection induces protective antitumor T cell immunity. J Immunol 171:3941-3946. (IF 5,52) PhD/MD students (last 5 years) and titles of their theses PhD Victor Huang, Rapid accumulation of epidermal CCL27 in skin-draining lymph nodes following topical application of a contact sensitizer recruits CCR10-expressing T cells, 2007-2008 (Institution: National Institutes of Health, Bethesda, USA) APPENDIX II MD Ellen Memaj, Untersuchung der Auswirkungen von UVB, UVA und UVA-1-Bestrahlung auf die Chemokinexpression humaner Keratinozyten, to date 95 PROF. DR. MED. KNUT SCHÄKEL Vice Chairmen Associate Professor for Immunodermatology Department of Dermatology Heidelberg University Hospital Ruprecht-Karls-University, Heidelberg Voßstr. 2 69115 Heidelberg 06221 56 8447 (Fon) 06221 56 8449 (Fax) [email protected] Curriculum vitae 2009 - to date 2009 2008 2005 2004 2003 2000 1995 - 2000 1993 - 1995 1993 1990 - 1991 Vice chairman, Department of Dermatology and Associate Professor (W3), Ruprecht-Karls-University Heidelberg Associate Professor (W2), Department of Dermatology, TU Dresden Consultant Immunologist, German Society of Immunology Habilitation in Immunology and Venia legendi in Dermatology Consultant, Department of Dermatology, TU Dresden Board certificate in Dermatology and Allergy Resident, Department of Dermatology, TU Dresden Postdoctoral Research Fellowship, Institute of Immunology, TU Dresden Resident, Department of Dermatology, Georg-August University of Göttingen Dissertation, Department of Pediatrics, Medical School Hannover Predoctoral Research Fellowship, DAAD, State University of New York, Buffalo, USA Areas of research expertise Immunology, Dendritic cells, Innate immunity APPENDIX II 5 selected (most important) publications Döbel, T., A. Kunze, J. Babatz, K. Tränkner, A. Ludwig, M. Schmitz, A. Enk and K. Schäkel. 2013. FcγRIII (CD16) equips immature 6-sulfo LacNAc-expressing dendritic cells (slanDCs) with a unique capacity to handle IgG-complexed antigens. Blood. 18:3609-18. (IF 9,89) Hansel, A., C.Gunther, J.Ingwersen, J.Starke, M.Schmitz, M.Bachmann, M.Meurer, E.P.Rieber, and K.Schäkel. 2011. Human slan (6-sulfo LacNAc) dendritic cells are inflammatory dermal dendritic cells in psoriasis and drive strong TH17/TH1 T-cell responses. J. Allergy Clin. Immunol. 127:787-794. (IF 11,0) Randolph, G.J., G.Sanchez-Schmitz, R.M.Liebman, and K.Schäkel. 2002. The CD16(+) (FcgammaRIII(+)) subset of human monocytes preferentially becomes migratory dendritic cells in a model tissue setting. J. Exp. Med. 196:517-527. (IF 13,38) Schäkel, K., R.Kannagi, B.Kniep, Y.Goto, C.Mitsuoka, J.Zwirner, A.Soruri, K.M.von, and E.Rieber. 2002. 6-Sulfo LacNAc, a novel carbohydrate modification of PSGL-1, defines an inflammatory type of human dendritic cells. Immunity. 17:289-301. (IF 21,63) Schäkel, K., K.M.von, A.Hansel, A.Ebling, L.Schulze, M.Haase, C.Semmler, M.Sarfati, A.N.Barclay, G.J.Randolph, M.Meurer, and E.P.Rieber. 2006. Human 6-sulfo LacNAc-expressing dendritic cells are principal producers of early interleukin-12 and are controlled by erythrocytes. Immunity. 24:767-777. (IF 21,63) PhD/MD students (last 5 years) and titles of their theses 96 PhD Annette Ebling, Die funktionelle Modifikation der proinflammatorischen M-DC8+ dendritischen Zellen durch zyklisches Adenosin-Monophosphat. magna cum laude Anja Hänsel, Programmierung von Th17-1 T-Zellen durch slan-dendritische Zellen und deren Bedeutung bei Autoimmunerkrankungen. magna cum laude Thomas Döbel, Immunregulatorische Bedeutung der Expression des Fc-gamma Rezeptors III bei humanen dendritischen Zellen. Stephanie Oehrl, Untersuchungen zur Funktion von Inflammasomen bei nativen humanen dendritischen Zellen MD Susanne Baumeister, Einfluss von G-CSF auf die Mobilisation und immunregulatorische Funktion von slan( 6-sulfo LacNAc+) – dendritischen Zellen, summa cum laude Matthias von Kietzell, Ausreifung und funktionelle Charakterisierung einer neuen Population von humanen dendritischen Zellen (slanDC), summa cum laude Jens Ingwersen, Funktioneller Vergleich von Subpopulationen dendritischer Zellen des menschlichen Blutes. summa cum laude Katja Rückert, Einfluss von TACE-Inhibitoren auf die Ausreifung proinflammatorischer M-DC8+ dendritischer Zellen. magna cum laude und Funktion Adele Heinrich: Einfluss des Anaphylatoxins C5a auf die Funktion proinflammatorischer 6-Sulfo LacNAc-exprimierender dendritischer Zellen (slanDCs) des humanen Blutes. magna cum laude Anke Döhring: Untersuchungen anhand eines neuen monoklonalen Antikörpers mit Spezifität für Fc gamma Rezeptor III (CD16+) exprimierende dendritische Zellen des menschlichen Blutes. magna cum laude Current extramural funding Untersuchungen zur Bedeutung von pro-inflammatorischen slan-dendritischen Zellen bei der Psoriasis, 2011 – 2015, Sonderforschungsbereich 938, Projekt N 97 APPENDIX II Sabine Seibel: Thema: Generierung 6-Sulfo LacNAc-exprimierender dendritischer Zellen (slanDCs) aus hämatopoetischen Vorläuferzellen. magna cum laude DR. MED. UNIV. ASTRID SCHMIEDER Specialist in Dermatology, Venereology, and Allergy Department of Dermatology, Venereology, and Allergy, University Medical Center Mannheim, Heidelberg University Theodor-Kutzer-Ufer 1-3 68135 Mannheim 0621 383 2280 (Fon) 0621 383 4085 (Fax) [email protected] Curriculum vitae since 2013 2013 2013 grant 2011 2011 2007 – 2013 2007 2007 2007 2005 2001 1999 – 2007 1999 Senior physician, University hospital Mannheim, Department of Dermatology Board certification Allergology Board certification Dermatology2012 Olympia Morata Gerok grant of the SFB 938: Milieuspezifische Kontrolle immunologischer Reaktivität Poster Award: Synergistic activation by p38MAPK and glucocorticoid signaling mediates induction of M2-like tumor-associated macrophages expressing the novel CD20 homolog MS4A8A Resident, University hospital Mannheim, Department of Dermatology Italian medical board exam Promotion to Dr. med. (Institute of pathophysiology at the Universitiy of Innsbruck, Austria) with the dissertation entiteled: Isogentisin- A novel compound for the prevention of smoking-caused endothelial injury Dr. Maria Schaumayer grant Otto Seibert merit grant Otto Seibert merit grant MD Studies in Innsbruck, University of Innsbruck, Austria Dante Alleghieri award APPENDIX II Areas of research expertise Tumor associated macrophages, Signal transduction, Dermatohistopathology, Psoriasis vulgaris 5 selected (most important) publications Michel J, Schonhaar K, Schledzewski K, Gkaniatsou C, Sticht C, Kellert B, Lasitschka F, Geraud C, Goerdt S, and Schmieder A. 2013. Identification of the novel differentiation marker ms4a8b and its murine homolog ms4a8a in colonic epithelial cells lost during neoplastic transformation in human colon. Cell Death Dis 4:e469 (IF 6.04) Schmieder A, Michel J, Schonhaar K, Goerdt S, and Schledzewski K. Differentiation and gene expression profile of tumor-associated macrophages. Semin Cancer Biol 22:289-297. (IF 7,44) Schmieder A*, Schledzewski K*, Michel J, Schonhaar K, Morias Y, Bosschaerts T, Van den Bossche J, Dorny P, Sauer A, Sticht C, Geraud C, Waibler Z, Beschin A, and Goerdt S. 2012. The cd20 homolog ms4a8a integrates pro- and anti-inflammatory signals in novel m2-like macrophages and is expressed in parasite infection. Eur J Immunol 42:2971-2982. (IF 4.97) Schmieder A*, Schledzewski K*, Michel J, Tuckermann JP, Tome L, Sticht C, Gkaniatsou C, Nicolay JP, Demory A, Faulhaber J, Kzhyshkowska J, Geraud C, and Goerdt S. 2011. Synergistic 98 activation by p38mapk and glucocorticoid signaling mediates induction of m2-like tumor-associated macrophages expressing the novel cd20 homolog ms4a8a. Int J Cancer 129:122-132. (IF 6.20) Schmieder A*, Schwaiger S*, Csordas A, Backovic A, Messner B, Wick G, Stuppner H, Bernhard D. 2007. Isogentisin--a novel compound for the prevention of smoking-caused endothelial injury. Atherosclerosis. 194:317-325. (IF 3,71) PhD students (last 5 years) and titles of their theses Michel, Julia; A comprehensive functional analysis of the CD20 homolog MS4A8A in macrophages and colonocytes (2010-2014). Schönhaar, Kathrin; Identification and characterization of progression-associated TAM molecules in malignant melanoma (2011-2014). Claudia, Dollt; Identification of signalling pathways essential for the development of TAM phenotypes in malignant melanoma (2013-2016). MD students (last 5 years) and titles of their theses Cleopatra, Gkaniatsou; Ms4a8a over-expression inhibits tumr growth of CT26 colon carcinoma cells in vivo (2009-2010). Cum laude Daniel, Behr; Charakterisation of the inflammatory infiltrate in the merkel cell carcinoma (20132014). APPENDIX II Current extramural funding SFB 938 (2011-2014), Projekt H, S. Goerdt: Tumor-assoziierte Makrophagen (TAM) und Tumorprogression: funktionelle Plastizität und progressionsabhängige TAM-Targetmoleküle beim Malignen Melanom 99 PROF. DR. MED. STEFAN WERNER SCHNEIDER Associate Professor for Clinical and Experimental Dermatology Section of Experimental Dermatology, Department of Dermatology, Venereology, and Allergy, University Medical Center Mannheim, Heidelberg University Theodor-Kutzer-Ufer 1-3 68135 Mannheim 0621 383 6902 (Fon) 0621 383 6903 (Fax) [email protected] Curriculum vitae Since 12/08 12/08 09/08 10/07 APPENDIX II 06/07 06/06 04/06 since 2005 04/05 08/02 03/02 07/01 since 11/00 03/99 11/97 10/97-06/02 08/97-09/97 08/96-09/97 08/94-07/96 05/95 07/94 05/94 11/89-02/94 11/87-05/94 Head of the Section “Experimental Dermatology” and Assistant Medical Director (“Leitender Oberarzt”) at the Department of Dermatology, Venereology and Allergology W3 “Cellular Differentation” University Mannheim-Heidelberg due the nomination of the Department of Dermatology Mannheim (Director: Prof. Dr. Sergij Goerdt) as the “Exzellenzzentrum Dermatologie in BadenWürttemberg”. specialized in phlebology Award for „innovative medical technology“ received from Federal Ministry of Education and Research (BMBF) by Ms Dr. A. Schavan Apl-Professur Senior supervisor at the Department of Dermatology, University of Münster Specialized in dermatology, allergy and venerology Development aid in Cambodia (regularly in Phnom Penh) Oskar-Gans-Award (Dermatology) Specialized in physiology Department of Dermatology, University of Münster (Prof. Dr. T. Luger) Habilitation (Venia legendi in Physiolgy) Head of working group „cell dynamics“ Bennigsen-Foerder-Award Approbation Department of Physiology, University Münster (Prof. Dr. H. Oberleithner) Fellowship (Anniversary award University Würzburg) at Department of Cellular and Molecular Physiology, Yale University, USA Dept. Internal Medicine University Hospital Würzburg (Prof. Dr. K. Wilms) DFG-Fellowship at Dept. of Physiology Univ. Würzburg (Prof. Dr. H. Oberleithner), Dept. of Surgery and Cellular Physiology, Yale University, USA (Prof. Dr. J. Geibel/Prof. Dr. G. Giebisch) Franconian Science Prize in Human Medicine Received Dr. of Medicine („Summa cum Laude“) Medical License at University of Würzburg Doctoral Thesis (Prof. Dr. H. Oberleithner, Physiology, Univ. of Würzburg) School of Medicine University Würzburg, Izmir (Turkey), Chur (CH) Areas of research expertise Mechanism of melanoma cell invasion and adhesion, Endothelial cell biology, von Willebrand factor, endothelium - tumor cell communication, mechanism of metastasis, new innovative medical technologies (5D-Intravitaltomography, micro-/nanofluidic, RICM, atomic force microscopy). 100 5 selected (most important) publications Görge, T., A.Barg, E.M.Schnäker, B.Pöppelmann, V.Shpacovitch, A.Rattenholl, T.A.Luger, M.Steinhoff, and S.W.Schneider. 2006. Melanoma-derived MMP-1 targets endothelial PAR1 promoting endothelial cell activation. Cancer Res. 66:7766-74. (IF 8,65) Pappelbaum, K.I., C.Gorzelanny, S.Grässle, J.Suckau M.W.Laschke, M.Bischoff, C.Bauer, M.Schorpp-Kistner, C.Weidenmaier, R.Schneppenheim, T.Obser, B.Sinha, and S.W.Schneider. 2013. Ultra-large von Willebrand factor fibers mediate luminal Staphylococcus aureus adhesion to an intact endothelial cell layer under shear stress. Circulation 128:50-59. (IF 15,20) Riehemann, K, S.W.Schneider, T.A.Luger, B.Godin, M.Ferrari, and H.Fuchs. 2009. Nanomedicine: challenge and oppurtunities. Angewandte Chemie, Int. edition 48:872-97, 2009. (wurde auch in deutscher Sprache publiziert) (IF 13,455) Kerk, N., E.A.Strozyk, B.Pöppelmann, and S.W.Schneider. 2010. The mechanism of melanomaassociated thrombin activity and von Willebrand factor release from endothelial cells. J Invest Dermatol. 130(9):2259-68. (IF 6,314) Schneider, S. W., S.Nuschele, A.Wixforth, A.Alexander-Katz, R.R.Netz, C.Gorzelanny, and M.F.Schneider. 2007. Shear-induced unfolding triggers adhesion of vWf fibers. Proc Natl Acad Sci USA 104:7899-903. (IF 9,74) PhD/MD students (last 5 years) and titles of their theses PhD Dr. rer. nat. Chrisitan Gorzelanny, Bioaktivität von Chitosan an Endothel-, Leukozyten und Tumorzellen, 2005-2008, Auszeichnung der Universität Münster Dipl.-biol. Anna Desch, Melanom-Endothel Kommunikation: Mechanismen der Endothelzellaktivierung und Bedeutung für die Metastasierung, 2009-andauernd MD Dr. med. Andre Niemeyer, Folgen der extrazelulärer Azidose an humanen Endothelzellen, 20002006 Dr. med. Afschim Fatemi, Effektive Therapie der Hyperhidrosis mittels Suctionscürettage, 20062007 Dr. med. Alexej Barg, Polymersierung des von-Willebrand Faktors in vitro, 2002-2007, Promotionspreis der Universität Münster Zahnärztin Katharina Podolewski, Die Bedeutung der ProteinaseActivated-Rezeptors (PAR) für die Melanomzellinvasion, 2004-2008 Arzt Felix Kleinrüschkamp, Mechanismen der Extravasation von Melanomzellen, Bedeutung des endothelialen von-Willebrand Faktors als Adhäsionsmolekül, 2005-2011 Dr. med. Nina Kerk, Melanom-Endothel Kommunikation: Aktivierung des endothelialen Weges und dessen Folge für die Expression von Adhäsionsmolekülen, 2007-2011 101 nFkB APPENDIX II Dipl.-biol. Karin Pappelbaum, Staphylokokken-Endothel Adhäsion und Endothelzellaktivierung: Bedeutung der Chitotriosidase, 2009-andauernd cand. med. Karin Roters, Wirkung von Pimecrolimus auf die Keratinozytenfunktion und – morphologie, 2007-andauernd cand. med. Margit Esser, Rasterkraftmikroskopische Untersuchungen nativer Alters- und Steroidgeschädigter Haut, 2007-andauernd cand. med. Verena Niemeyer, Flußbedingungen, 2009-andauernd Melanomzelladhäsion am Endothel unter definierten cand. med. Jan Suckau, ULVWF in Tumorgefäßen der Maus und Mensch, 2010-andauernd cand. Med. Sarah Schober; Rutin und Vitamin C als effective Therapie der Purpura Pigmentosa Progressiva, 2011-andauernd cand. med. Lukas Görtz, Melanom-Endothelzell Kommunikation im lymphatischen System, 2011andauernd Current extramural funding SFB/TR 23 (TP A9) Vascular differentiation and remodelling “Mechanism controlling the transition from quiescent to activated endothelium cells mediated by tumor cells”, 07.2013-06.2017 BMBF „Anwendung der Intravitaltomographie (Woundoptomizer) an chronischen Wunden”, 10.2010-12.2013 DFG Wissenschaftliche Forscherguppe „Shear flow regulated hemostasis“, (SCHN 474/5-1); 10.2011 bis 09.2014 EU Verbundprojekt „Nano3Bio (Nr. 616931)“, 10.2013-09.2017 APPENDIX II 102 PROF. DR. JONATHAN P. SLEEMAN Franz-Volhard-Stiftungsprofessur für Mikrovaskuläre Biologie und Pathobiologie (W3) University Medical Center Mannheim, Heidelberg University Ludolf-Krehl-Str 13-17 68167 Mannheim 0621 383 9955 (Fon) 0621 383 9961 (Fax) [email protected] Curriculum vitae 2008–to date: W3-Franz-Volhard-Stiftungsprofessur für Mikrovaskuläre Biologie und Pathobiologie, Universität Heidelberg (Medizinische Fakultät Mannheim) 2008–to date: Group leader, Karlruhe Institute of Technology, Institut für Toxikologie und Genetik (secondary employment basis) 2004 - 2007: Acting Chair of Genetics (W3), Institut für Genetik, University of Karlsruhe 2002 – 2007 Deputy Director, Forschungszentrum Karlsruhe, Institut für Toxikologie und Genetik 2000 Habilitation (Fach: Genetik), University of Karlsruhe 1997 - 2007: Group leader, Forschungszentrum Karlsruhe, Institut für Toxikologie und Genetik 1992-1996: Postdoctoral studies, Forschungszentrum Karlsruhe, Institut für Genetik EMBO long-term fellowship (1992-1993) Marie Curie Fellow of the European Union (1994-1996) 1987-1991: PhD degree, Trinity College, Cambridge University, England 1984-1987: 1st Class Honours Degree in Natural Sciences, specialising in Biochememistry. Trinity College, Cambridge University, England 1983-1984: „Break Year Student“, Department of Cancer Studies, University of Birmingham, England 5 selected (most important) publications Sleeman, J.P., Rudy, W., Hofmann, M., Moll, J., Ponta, P. and Herrlich, P. 1996. Regulated clustering of variant CD44 proteins increases their hyaluronate binding capacity. J. Cell Biol., 135: 1139-1150. (IF 10.82) Thiele, W., Krishnan, J., Rothley, M., Weih, D., Plaumann, D., Kuch, V., Quagliata, L., Weich, H., Pytowski, B. and Sleeman, J. P. 2012. VEGFR-3 is expressed on megakaryocyte precursors in the murine bone marrow and plays a regulatory role in megakaryopoiesis. Blood, 120: 1899-1907. (IF 9.06) Nestl, A., Von Stein, O., Zatloukal, K., Thies, W.-G., Herrlich, P., Hofmann, M and Sleeman, J. P. 2001. Gene expression patterns associated with the metastatic phenotype in rodent and human tumors. Cancer Research, 61: 1569-1577. (IF 8.65) Krishnan, J., Kirkin, V., Steffen, A., Hegen, M., Weih, D., Tomarev, S., Wilting, J. and Sleeman, J. P. 2003. Differential in vivo and in vitro expression of VEGF-C and VEGF-D in tumors and its relationship to lymphatic metatasis in immunocompetent rats. Cancer Research, 63: 713-722 (IF 8.65) Baumann, P., Cremers, N., Kroese, F., Orend, G., Chiquet-Ehrismann, R., Uede, T., Yagita, H. and Sleeman, J. P. 2005. CD24 expression causes the acquisition of multiple cellular properties associated with tumor growth and metastasis. Cancer Research, 65: 10783-10793 (IF 8.65) 103 APPENDIX II Areas of research expertise Metastasis, lymphangiogenesis PhD students (last 5 years) and titles of their theses 2010 Nicole Grau. Zur Rolle von Lymphknotenmetastasen in der Metastasierung von Tumoren in lebenswichtige Organe. Note: 1,0 (sehr gut) 2011 Anja Schmaus. Regulation der Lymphangiogenese: Moleküle, Mechanismen und die Rolle in pathologischen Prozessen. Note: 1,0 (sehr gut), Summa cum lauda 2011 Luca Quagliata. Local, regional and systemic roles of VEGF-C in tumor metastasis. Note: Magna cum lauda 2012 Jochen Bauer. Die Rolle niedermolekularer Hyaluronsäurefragmente und LYVE-1 bei der Lymphangiogenese. Note: Magna cum lauda 2012 Vanessa Kuch. Evaluierung und Identifizierung zuverlässiger Methoden zur Isolierung von Krebsstammzellen. Note: Summa cum lauda 2013 Anna Poletti. Protein interaction partners of ASAP1 and their role in metastasis. Note: Magna cum lauda Current PhD students: Haniyeh Sabouri, Justyna Krachulec, Sandra Klusmeier, Supriya Saraswati, Lisa Jerabek Current extramural funding HGF Biointerfaces Joint/Twinning Programme. Cell-free metabolic cascades for the enzymatic synthesis of tailor-made oligosaccharides using microfluidic devices (together with PD Dr. Ute Schepers and Prof. Dr. Matthias Franzreb) Baden-Württemberg Stiftung Forschungsprogramm „Glykobiologie / Glykomik“ Consortium: „Glykobiologie von Hyaluronsäure-Oligosacchariden: biologische und klinische Relevanz“. Coordinator: Jonathan Sleeman EU Interreg Consortium „Nanomatrix“ Nanoparticles for imaging the lymphatics. Coordinator: Genevieve Pourroy (France) APPENDIX II DAAD PhD Stipendium Wilhelm Sander-Stiftung The functional interplay between CD24, Src, miR-21, Pdcd4 and u-PAR, and its impact on invasion and metastasis (together with Prof. Dr. Heike Allgayer). German-Israeli Foundation (GIF) The role of VEGF-C in the mobilization of immune cells following anti-cancer drug therapy, and their impact on tumor re-growth and metastasis (together with Yuval Shaked) 104 PROF. DR. VIKTOR UMANSKY Group leader, Clinical Cooperation Unit Dermato-Oncology, German Cancer Research Center (DKFZ) and Department of Dermatology, Venereology, and Allergy, University Medical Center Mannheim, Ruprecht-Karl University of Heidelberg Theodor-Kutzer-Ufer 1-3 68135 Mannheim 0621 383 3373 (Fon) 0621 383 2163 (Fax) [email protected] 1972-1978 Study of medicine at the State Medical Institute, Kiev, Ukraine 1982 Ph.D. degree in “Experimental Oncology and Immunology”, Institute for Oncology, Kiev 1991 Doctor of Science degree in “Experimental Oncology and Immunology”, Institute for Oncology, Kiev 2001 Habilitation (Venia Legendi) in “Pharmacological Biology” and the acquisition of the Title “Privatdozent”, Faculty for Phramacology, University Heidelberg, Heidelberg, Germany 2008 Acquisition of the Title “Professor”, Faculty for Biosciences, University Heidelberg 1979-1982 PhD Student. Department of Molecular Immunology, Institute of Biochemistry, Kiev, Ukraine 1983-1986 Research associate. Department of Tumor Metastasis, Institute for Oncology, Kiev 1986-1988 Senior research associate. Department of Tumor Metastasis, Institute for Oncology, Kiev 1988-1991 Leader of the group “Immune Antitumor Resistance”. Department of Tumor Metastasis, Institute for Oncology, Kiev. 1991-1992 Postdoc. Laboratory of Immunology, INSERM U.252, Dijon, France. 1992-1994 Postdoc. Department of Cellular Immunology, Tumor Immunology Program, German Cancer Research Center (DKFZ), Heidelberg, Germany 1994-1996 Research associate. Department of Cellular Immunology, DKFZ. 1996-1998 Senior research associate. Department of Cellular Immunology, DKFZ. 1998-2001 Leader of the group “Tumor Immunotherapy” Department of Cellular Immunology, DKFZ 2001-2002 Leader of the laboratory of Immunology. Company Virofem Diagnostica, Wiesbaden, Germany 2002-present Leader of the group “Mouse Models of Spontaneous Melanoma for Immunotherapy”. Skin Cancer Unit, DKFZ and University Hospital Mannheim, Germany 2002 Sir Hans Krebs Prize 2001 for the outstanding work in the biomedical science entitled “Therapy of human tumors in NOD/SCID mice with patient derived reactivated memory T cells from bone marrow“ and published in Nature Medicine 2001, 7: 452-458 105 APPENDIX II Curriculum vitae Areas of research expertise Tumor immunotherapy, immunosuppression, suppressor cells, effector and regulatory T cells cancer and inflammation, myeloid-derived 5 selected (most important) publications (Impact Factor 2012) Meyer, C., A. Sevko, M. Ramacher, A.V. Bazhin, C.S. Falk, W. Osen, I. Borrello, M. Kato, D. Schadendorf, M. Baniyash, V. Umansky. 2011. Chronic inflammation promotes myeloid derived suppressor cell activation blocking antitumor immunity in transgenic mouse melanoma model. Proc. Natl. Acad. Sci. USA, 108: 17111-17116. (IF = 9.74) Umansky, V., O. Abschuetz, W. Osen, M. Ramacher, F. Zhao, M. Kato, D. Schadendorf. 2008. Melanoma specific memory T cells are functionally active in ret transgenic mice without macroscopical tumors. Cancer Res. 68: 9451-9458. (IF = 8.65). Beckhove, P., M. Feuerer, M. Dolenc, F. Schuetz, C. Choi, N. Sommerfeldt, J. Schwendemann, K. Ehlert, P. Altevogt, G. Bastert, V. Schirrmacher, V. Umansky. 2004. Specifically activated memory T cell subsets from cancer patients recognize and reject xenotransplanted autologous tumors. J. Clin. Invest. 114: 67-76. (IF = 12.81) Feuerer, M., P. Beckhove, N. Garbi, Y. Mahnke, A. Limmer, M. Hommel, G. Hämmerling, B. Kyewski, A. Hamann, V. Umansky, V. Schirrmacher. 2003. Bone marrow as a priming site for Tcell responses to blood-borne antigen. Nature Med. 9: 1151-1157. (IF = 22.86) Feuerer, M., P. Beckhove, L. Bai, E.F. Solomayer, G. Bastert, I.J. Diehl, C. Pedain, M. Oberniedermayr, V. Schirrmacher, V. Umansky. 2001. Therapy of human tumors in NOD/SCID mice with patient derived re-activated memory T cells from bone marrow. Nature Med. 7: 452-458. (IF = 22.86) PhD/MD students (last 5 years) and titles of their theses PhD APPENDIX II Abschütz, Oliver. Melanoma antigen specific memory T cells in transgenic mouse model of spontaneous melanoma, cum laude, 2005-2008. Zhao, Fang. Tolerogenic dendritic cells in ret transgenic mouse model of spontaneous melanoma, magna cum laude, 2006-2009. Kimpfler, Silvia. Characterization of CD4+CD25+FOXP3+ regulatory T cells in ret transgenic mouse melanoma model and in melanoma patients, magna cum laude, 2006-2010. Meyer, Christiane. Effect of myeloid-derived suppressor cells on the expression of T cell receptor ζ-chain and on melanoma development in ret transgenic mouse model, magna cum laude, 20062010. Flores-Guzman, Fernando. Dormant tumor cells in ret transgenic mouse melanoma model and their interaction with T cells, magna cum laude, 2008-2012. Stemke, Anastasia. Immunotherapy with tumor antigen-specific T cells in ret transgenic mouse melanoma model, cum laude, 2009-2013. Ivan Shevchenko. Extracellular adenosine metabolism in melanoma and pancreatic cancer, magna cum laude, 2010-2013. 106 Current extramural funding In situ ablation of primary tumors to induce anti-tumor T-cell reactions and neutralize immunosuppresive tumor microenvironment. 2012-2014. German-Israeli Foundation for Scientific Research and Development (GIF). The role of CCR5 in the recruitment of myeloid-derived suppressor cells (MDSC) from the bone marrow to support melanoma progression. 2013-2016. DKFZ/MOST Cooperation Program in Cancer Research. APPENDIX II Combined immunotherapy of malignant melanoma using dendritic cell vaccination and neutralization of immunosuppression in pre-clinical mouse models. 2014-2017. German-Israeli Helmholtz Research School in Cancer Biology (DKFZ-WIS). 107 PROF. DR. MED. JOCHEN UTIKAL Head of the Clinical Cooperation Unit Dermato-Oncology, German Cancer Research Heidelberg (DKFZ) and Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, University of Heidelberg Theodor-Kutzer-Ufer 1-3 68135 Mannheim +49-(0)621-383-4461 (Fon) +49-(0)621-383-3815 (Fax) [email protected] Curriculum vitae since 3/2012 2009-2012 12/2009 2007-2009 2007 2007 2006 2002-2006 2001 2001 1994-2001 Head of the Clinical Cooperation Unit Dermato-Oncology , German Cancer Research Center (DKFZ) and Dept. of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Ruprecht-Karl University of Heidelberg Attending (Oberarzt) and Research Group Leader, Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Ruprecht-Karl University of Heidelberg Venia legendi for Dermatology and Venereology, Ruprecht-Karl University of Heidelberg Research Fellowship Massachusetts General Hospital, Cancer Center and Harvard Stem Cell Institute, Boston Medical board certification “Medical Tumor Therapy“ Medical board certification “Allergology” Medical board certification “Dermatology and Venereology” Resident in Dermatology, Department of Dermatology, Venereology and Allergology, University Medical Center Mannheim, Ruprecht-Karl University of Heidelberg Resident, Dept. of Dermatology, University of Ulm Doctoral thesis: The significance of the c-myc and bcl-2 gene in the pathogenesis of malignant melanoma “summa cum laude” Study of Medicine at the Universities of Ulm and Berne APPENDIX II Areas of research expertise Translational Dermato-Oncology, Malignant Melanoma, Stem Cells Honors and Awards (since 2008): 2011 Fleur-Hiege Memorial Award 2010 Hella Bühler-Award for Cancer Research of the Ruprecht-Karl University of Heidelberg 2010 Egon Macher Award 5 selected (most important) publications Hirata A*, Utikal J*, Yamashita S, Aoki H, Watanabe A, Yamamoto T, Okano H, Bardeesy N, Kunisada T, Ushijima T, Hara A, Jaenisch R, Hochedlinger K, Yamada Y. 2013. Dose-dependent roles for canonical Wnt signalling in de novo crypt formation and cell cycle properties of the colonic epithelium. Development. 140: 66-75. *authors contributed equally (IF 6,20) Flaherty KT, Robert C, Hersey P, Nathan P, Garbe C, Milhem M, Demidov LV, Hassel JC, Rutkowski P, Mohr P, Dummer R, Trefzer U, Larkin JM, Utikal J, Dreno B, Nyakas M, Middleton MR, Becker JC, Casey M, Sherman LJ, Wu FS, Ouellet D, Martin AM, Patel K, Schadendorf D. 2012. Improved Survival with MEK Inhibition in BRAF-Mutated Melanoma. N Engl J Med. 367:10714. (IF 51,65) 108 Utikal J, Polo JM, Stadtfeld M, Maherali N, Kulalert W, Walsh RM, Khalil A, Rheinwald JG, Hochedlinger K. 2009. Immortalization eliminates a roadblock during the reprogramming of somatic cells into iPS cells. Nature 460:1145-8. (IF 38,59) Utikal J, Maherali N, Kulalert W, Hochedlinger K. 2009. Sox2 is dispensable for the reprogramming of melanocytes and melanoma cells into induced pluripotent stem cells. J Cell Sci 122:3502-10. (IF 5,87) Stadtfeld M, Nagaya M, Utikal J, Weir G, Hochedlinger K. 2008. Induced pluripotent stem cells generated without viral integration. Science 322:945-9. (IF 31,02) PhD/MD students (last 5 years) and titles of their theses PhD Bernhardt, Mathias; Reprogramming of melanoma cells into a pluripotent state. 2011-2014 Galach, Marta; Dissection of molecular mechanisms of keratinocyte differentiation from iPS cells. 2010-2014 Korona, Danuta; Generation of tumor-antigen-specific T cells from induced pluripotent stem cells (iPS cells) for melanoma immunotherapy. 2013-2016; together with Viktor Umansky Schöler, Nathalie; Melanoma stem cell markers and epigenetic plasticity. 2012-2015 Weina, Kasia; Differentiation mechanisms of iPS cells into melanocytes. 2012-2015 MD Lichtenberger, Ramtin; Biomarker of malignant melanoma, 2012-2014 Current extramural funding German research council (DFG, SFB 873, A-8, PI 2010-2014 and SFB636, B-07, PI 2012-2015) Hella Bühler-Award 2010 Baden-Württemberg Foundation (Adult stem cells II, PI) 2010- 2015 German Cancer Aid, Max-Eder Research Group 2011-2015 BMBF, DZHK 2012-2015 109 APPENDIX II Orouji, Elias; Diagnostic markers of malignant melanoma, 2011-2014 PROF. DR. MED. FRANK WINKLER Consultant Dpt.of Neurooncology Neurology Clinic and National Center for Tumor Diseases Im Neuenheimer Feld 400 69120 Heidelberg 06221 56 37772 (phone) 06221 56 5935 (fax) [email protected] Curriculum vitae since 4/2012 W3 Professorship “Experimental Neurooncology”, University of Heidelberg since 2010 Consultant and Assistant Professor, Dpt. of Neurooncology, University of Heidelberg Establishment and PI of the research group “Experimental Neurooncology” 2010 Venia Legendi for Neurology, University of Munich (2011: University of Heidelberg) 2009 Boards in Neurology 2005-2010 Resident, Dpt. of Neurology, University of Munich, Großhadern clinic. Establishment and PI of the research group “Experimental Neurooncology”. 2003 and 2004 Research Fellow, Steele Lab, Harvard University, Boston (Rakesh Jain) 1999-2002 Resident and Postdoctoral studies, Dpt. of Neurology, University of Munich, Großhadern clinic 1998 Doctoral thesis (degree Dr. med., Dpt. of Cardiology, Univ. Freiburg), and Approbation 1994 – 1998 MD studies, University of Freiburg 1991-1994 MD studies, University of Hamburg 2002 2010 Scientific award of the Paul-Ehrlich-Gesellschaft Sibylle-Assmus Award for Neurooncology APPENDIX II Areas of research expertise Neuro-oncology; brain metastasis of melanoma and lung carcinoma; in vivo imaging, angiogenesis, invasion 5 selected (most important) publications von Baumgarten L, Brucker D, Tirniceru A, Kienast Y, Grau S, Burgold S, Herms J, Winkler F (2011). Bevacizumab has differential and dose-dependent effects on glioma blood vessels and tumor cells. Clin Cancer Res 17:6192-205. (IF 7.8) Kienast Y, von Baumgarten L, Fuhrmann M, Klinkert W, Goldbrunner R, Herms J, F Winkler. 2010. Real-time imaging reveals the single steps of brain metastasis formation. Nature Med 16: 116-122. (IF 24.3) Winkler F, Kienast Y, Fuhrmann M, von Baumgarten L, Burgold S, Mitteregger G, J Herms. 2009. Imaging glioma cell invasion in vivo reveals mechanisms of dissemination and peritumoral angiogenesis. Glia 57:1306-15. (IF 5.1) Winkler F, Kozin SV, Tong RT, Chae S, Booth MF, Garkavtsev I, Xu L, Hicklin DK, Fukumura D, di Tomaso E, Munn LL, RK Jain RK. 2004. Kinetics of vascular normalization by VEGFR2 blockade governs brain tumor response to radiation: Role of oxygenation, Angiopoietin-1, and matrix metalloproteinases. Cancer Cell 6: 553-563. (IF 24.8) 110 Garkavtsev I, Kozin SV, Chernova O, Xu L, Winkler F, Brown E, Barnett GH, RK Jain. 2004. The candidate tumour suppressor protein ING4 regulates brain tumour growth and angiogenesis. Nature 428: 328-32. (IF 38.6) PhD/MD students (last 5 years) Yvonne Kienast, PhD: “The mechanisms of brain metastasis formation: a new experimental approach via in vivo two-photon microscopy”. 2005-2009. Summa cum laude. Gergely Solecki, PhD student: “Optimization of vascular normalization by anti-VEGF-A and antiAng-2 therapy for improved chemotherapy and radiotherapy”. Started 11/2012. Mustafa Syed, MD student: “Calcium communication between glioma cells in vitro and in vivo”. Started 1/2013. Erik Jung, MD student: “Inhibiting intercellular communication in brain tumors for improved antitumor therapies”. Started 2/2014 Kianush Karimian, MD student: “Advanced fluorescent reporter systems for dynamic detection of intratumoral heterogeneity in brain tumor progression”. Started 11/2013 Current extramural funding Die Bedeutung der Tumor-Stammzellen für die einzelnen Schritte der Gehirnmetastasierung. 2011-2014. Deutsche Forschungsgemeinschaft, Projekt WI 1930/5-1 Die Wirkung der HER2-Expression, und anti-HER2/-VEGF Therapien auf die einzelnen Schritte der Hirnmetastasierung von Brustkrebszellen. 2009-2012. Deutsche Krebshilfe, Projekt 109051. APPENDIX II Glioma Angiogenesis collaboration. 2012-2014. Roche, Deutschland, Euro 300.000 111 Appendix II 2. Biographical Sketches of the Associated Researchers PROF. DR. MED. CLAUS-DETLEV KLEMKE Consultant, Dept. of Dermatology, Venereology, and Allergy, University Medical Center Mannheim, Heidelberg University Theodor-Kutzer-Ufer 1-3 68135 Mannheim 0621 383 2280 (Fon) 0621 383 3815 (Fax) [email protected] APPENDIX II Curriculum vitae 1989 „Abitur“ (high school degree) 1989-1991 Compulsory Military Service 1991-1998 Medical studies at the Universities of Würzburg, London and Berlin 1998 „Ärztliche Prüfung“ (National Medical Examination); „Approbation“ (Medical board certification as a physician) 1999-2000 Resident, Dept. of Dermatology, Free University of Berlin 2000 Doctoral Certification, University of Würzburg, „magna cum laude“ 2000-2003 Resident, Dept. of Dermatology, Venereology, and Allergology, University Medical Center Mannheim, University of Heidelberg 2003 „Facharzt für Haut- und Geschlechts-krankheiten“ (Medical board certification for dermatology and venereology) 2004-2006 Postdoc in the Division of Immunogenetics, Tumour Immunology Program, German Cancer, Research Center (DKFZ), Heidelberg, Germany 2006 „Zusatzbezeichnung für Allergologie“ (Medical board certification for allergic diseases) 2007 „Zusatzbezeichnung Medikamentöse Tumortherapie“ (Medical board certification for medical tumor therapy (chemotherapy)) Since 2007 Speaker of the German working group on cutaneous lymphomas of the ADF (Arbeitsgemeinschaft Dermatologische Forschung) 2010 Habilitation Dermatologie und Venerologie 2011 „Zusatzbezeichnung Dermatohistologie“ (Medical board certification for dermatopathology) 2013 Ernennung zum außerplanmässigen Professor der Universität Heidelberg Areas of research expertise Dermato-Oncology, Immunology, Apoptosis 5 selected (most important) publications Kießling MK, Oberholzer PA, Mondal C, Karpova MB, Zipser MC, Lin WM, Girardi M, MacConaill LE, Kehoe SM, Hatton C, French LE, Garraway LA, Polier G, Klemke CD, Krammer PH, Gülow K, Dummer R. High-throughput mutation profiling and next-generation sequencing of CTCL samples reveal KRAS and NRAS mutations which sensitize tumors towards treatment with inhibitors targeting the RAS/RAF/MEK signaling cascade. Blood. 2011;117:2433-2440. (IF 10.56) Steininger A, Möbs M, Ullmann R, Köchert K, Kreher S, Anagnostopoulos I, Hummel M, Richter J, Beyer M, Janz M, Klemke CD, Stein H, Dörken B, Sterry W, Schrock E, Mathas S, Assaf C. Genomic loss of the putative tumor suppressor gene E2A promotes lymphoma in human. J Exp Med. 2011;208:1585-93. (IF 14.78) 112 Klemke CD*, Brenner D*, Weiss EM, Schmidt M, Leverkus M, Gülow K, Krammer PH. The lack of T cell receptor induced signaling is crucial for CD95 ligand up-regulation and protects cutaneous T cell lymphoma cells from Activation-Induced Cell Death. Cancer Res. 2009;69:4175-4183, *”equal contribution author” (IF 8.23) Heid JB, Schmidt A, Oberle N, Goerdt S, Krammer PH, Suri-Payer E*, Klemke CD*. FOXP3+CD25tumor cells with regulatory function in Sézary Syndrome. J Invest Dermatol. 2009;129:2875-85, *”equal contribution author” (IF 6.27) Klemke CD*, Fritzsching B*, Franz B, Kleinmann E, Oberle N, Poenitz N, Sykora J, Banham AH, Roncador G, Goerdt S, Krammer PH, Suri-Payer E. Paucity of FOXP3+ cells in skin and peripheral blood distinguishes Sézary Syndrome from other cutaneous T cell lymphomas. Leukemia 2006;20:1123-9., *”equal contribution author” (IF 8.97) PhD/MD students (last 5 years) and titles of their theses MD Julia Heid, Charakterisierung regulatorischer T Zellen beim Kutanen T Zell Lymphom, 2006 - 2008, summa cum laude, together with Prof. Dr. P. H. Kramer Melanie Faust; Analyse der Bedeutung regulatorischer T-Zellen bei kutanen Arzneimittelreaktionen, 2006 – 2013 Joana Schmidt; Auswertung zur klinischen Relevanz von allergologischen Testungen bei Unverträglichkeitsreaktionen auf Arzneimittel, 2011 – 2013 Jasmin Hambsch, Analyse der Behandlungsparameter bei der Extrakorporalen Photopherese, 2011 – 2013 Daniela Gräf, Retrospektive Auswertung zur Strahlentherapie kutaner Lymphome, 2012-2013. 113 APPENDIX II Christine Stumpf; Analyse der Expression von Angiopoietin-2 relevanter Proteine in primär kutanen BZell Lymphomen, 2011 – 2013, together with Dr. M. Felcht PROF. DR. MED. WIEBKE KATHARINA LUDWIG-PEITSCH Consultant (“Oberärztin”) Dept. of Dermatology, Venereology and Allergy, University Medical Center Mannheim, Heidelberg University Theodor-Kutzer-Ufer 1-3 68135 Mannheim Germany Phone: +49-621-383 1054 Fax. +49-621-383 3815 Email: [email protected] Curriculum vitae Since 06/2012 07/2012 Since 07/2011 Since 10/2010 Since 09/2009 07/2008 02/2008 02/2006-08/2008 07/2005 04/2003 03/2002-01/2006 APPENDIX II 03-12/2002 09/2000-02/2002 05/2000 10/1993-05/2000 06/1993 Grants 04/2004-05/2007 04-12/1999 03/1994-07/2000 Awards 10/2007 06/2007 06/2001 06/1998 Professor of Dermatology and Venereology, Medical Faculty Mannheim, Heidelberg University Specialist in Medical Tumor Therapy Head of the Division of Allergy, Occupational and Environmental Medicine, Department of Dermatology, University Medical Center Mannheim, Heidelberg University Deputy Head of the Outpatient Clinic, Department of Dermatology, University Medical Center Mannheim Consultant ("Oberärztin"), Department of Dermatology, University Medical Center Mannheim Postdoctoral Lecture Qualification ("Habilitation") in Dermatology and Venereology, Medical Faculty Mannheim, Heidelberg University Specialist in Allergy Senior Physician ("Funktionsoberärztin"), Department of Dermatology, University Medical Center Mannheim Specialist in Dermatology and Venereology (“Fachärztin”) Dissertation in Medicine, University of Hamburg in cooperation with the Division of Cell Biology, German Cancer Research Center, Heidelberg (grade: summa cum laude) Senior Resident ("Assistenzärztin"), Department of Dermatology, University Medical Center Mannheim Research Appointment, Division of Cell Biology (Head: Prof. Dr. rer. nat. Werner W. Franke), German Cancer Research Center, Heidelberg Junior Resident ("Ärztin im Praktikum"), Department of Dermatology and Venereology, University Hospital Eppendorf, Hamburg, Germany Medical State Exam (grade: 1.5) Medical studies, Heidelberg University, Germany; Harvard Medical School, Boston, MA, USA; University of Texas, Texas, USA A-levels, Goetheschule, Essen, Germany (grade: 1.0) Olympia-Morata Scholarship, Heidelberg University Grant by the German Academic Exchange Service (DAAD) Grant by the German National Merit Foundation (“Studienstiftung des deutschen Volkes”) Award for Experimental Research in Oncology, Working Group for Oncology (“Onkologischer Arbeitskreis“, OAK), Medical Faculty Mannheim, Heidelberg University Karl-Freudenberg-Award of the Heidelberg Academy of Sciences Award for publications by junior scientists from Northern German Dermatologic Hospitals, Lübeck, Germany Student Award, Benjamin Franklin Contest, Berlin, Germany 114 Areas of research expertise Dermato-Oncology (Malignant Melanoma, Merkel Cell Carcinoma), Cell Adhesion, Actin-binding Proteins, Psoriasis 5 selected (most important) publications Schrama D, Peitsch WK, Zapatka M, Kneitz H, Houben R, Eib S, Haferkamp S, Moore PS, Shuda M, Thompson JF, Trefzer U, Pföhler C, Scolyer RA, Becker JC. 2011. Merkel cell polyomavirus status is not associated with clinical course of merkel cell carcinoma. J Invest Dermatol 131:16318. (IF: 6.19) Schmitt CJ, Franke WW, Goerdt S, Falkowska-Hansen B, Rickelt S, Peitsch WK. 2007. Homoand heterotypic cell contacts in malignant melanoma cells and desmoglein 2 as a novel solitary surface glycoprotein. J Invest Dermatol 127:2191-2206. (IF: 6.19) Peitsch WK, Hofmann I, Bulkescher J, Hergt M, Spring H, Bleyl U, Goerdt S, Franke WW. 2005. Drebrin, an actin-binding, cell-type characteristic protein: induction and localization in epithelial skin tumors and cultured keratinocytes. J Invest Dermatol 125:761-74. (IF: 6.19) Peitsch WK, Hofmann I, Endlich N, Prätzel S, Kuhn C, Spring H, Gröne HJ, Kriz W, Franke WW. 2003. Cell biological and biochemical characterization of drebrin complexes in mesangial cells and podocytes of renal glomeruli. J Am Soc Nephrol 14:1452-63. (IF: 8.98) Kontoyiannis DK, Peitsch WK, Reddy BT, Whimbey EE, Han XY, Bodey G., Rolston KV. Cryptococcosis in patients with cancer. Clin Infect Dis 2001; 32:E145-150. (IF: 9.37) MD students and titles of their theses Schmitt, Christian; Homo- and heterotypic melanoma cell contacts and desmoglein 2 as a novel solitary cell surface protein, 2005-2008, summa cum laude, together with Prof. Dr. S. Goerdt. Werling, Anna Maria; Homo- and heterotypic cell contacts of Merkel cells and Merkel cell carcinomas: surprising heterogeneity and indications for a cadherin switch, 2008-2010, magna cum laude. Warnecke, Christine; Cardiovascular and metabolic comorbidities in psoriasis: a case-control study, 2009-2012, magna cum laude. Schaarschmidt, Marthe-Lisa; Conjoint analysis: a novel method for identification of patient preferences for psoriasis treatments, 2009-2012, magna cum laude, together with Dr. A. Schmieder. Vlahova, Lyubomira; P-cadherin: a novel prognostic marker in Merkel cell carcinomas, since 2010, under review. Schober, Sarah; Antioxidants as novel treatment approach in pigmented purpuric dermatosis: a retrospective case series, since 2010, together with Prof. Dr. S. W. Schneider. Poppe, Manuel; Impact of fumaric acid esters on cardiovascular and metabolic risk factors in psoriasis, since 2011, together with Dr. A. Schmieder. Martin, Isabelle; Patient preferences for treatment of basal cell carcinomas: a conjoint analysis, since 2012, together with Dr. A. Schmieder. Glocker, Anne; Surgical and conservative treatment of basal cell carcinomas: correlation of patient preferences with subjective and objective success, since 2012, together with Dr. A. Schmieder. 115 APPENDIX II Lang, Sabine; Inpatient and outpatient treatment of psoriasis – a health economic cost analysis from the societal perspective, 2006-2009, cum laude, together with Prof. Dr. M. Goebeler. Kromer, Christian; Patient preferences for treatment of psoriasis with biologicals, since 2012, together with Dr. M.-L. Schaarschmidt and Dr. A. Schmieder. Extramural funding Desmoglein 2 in malignant melanomas: cellular functions, diagnostic and prognostic significance, 2008-2011, project 108626, German Cancer Aid (“Deutsche Krebshilfe”). Topogenic and regulatory principles of the actin-binding protein drebin, 2006-2008, project PE 896/1-3, German Research Foundation (“Deutsche Forschungsgemeinschaft“). Molecular characteristics, complexes and functions of the actin-binding protein drebrin in nonneuronal cells, 2003-2006, projects PE 896/1-1 and 1-2, German Research Foundation. APPENDIX II 116 PROF. DR. RER. NAT. KARSTEN MAHNKE University Hospital Heidelberg Department of Dermatology Ruprecht Karls University Heidelberg Voßstraße 2 69115 Heidelberg Phone: +49 6221 56 8170 Fax +49 6221 561617 [email protected] Curriculum Vitae 2004 - to date Apl. Prof. Head of Research Laboratory for Dermatology 2003 Venia Legendi in Immunology (Habilitation, PD), University of Mainz, Mentor Prof. Dr. A. H. Enk 2001 - 2004 Postdoctoral Assistant (C1), Department of Dermatology, University of Mainz, Prof. A.H. Enk 1996 - 2001 Research Assistant “Laboratory of Cellular Physiology and Immunology”, The Rockefeller University New York, USA, Prof. R.M. Steinman. 1994 - 1996 Postdoc in the Department of Dermatology, University of Münster, Prof. T. Schwarz 1990 - 1994 Doctoral thesis in Immunology, Dr. rer. nat, Institute for Dermatological Research, University of Münster, Prof. C. Sorg 1990 Diploma Thesis in Neurophysiology, University of Münster, Prof E. Speckmann 1983 - 1989 Study of Biology, at the Universities Bremen, Oldenburg, and Münster, Germany 5 selected (most important) publications Ring S, Enk AH, Mahnke K. 2011. Regulatory T cells from IL-10-deficient mice fail to suppress contact hypersensitivity reactions due to lack of adenosine production. J Invest Dermatol. 131:1494-502. (IF 6,2) Ring S, Karakhanova S, Johnson T, Enk, A.H. and Mahnke K. 2010. Gap junctions between regulatory T cells and dendritic cells prevent sensitization of CD8+ T cells. J. Allergy Clin. Immunol. 125:237-247. (IF 10,3) Ring S, Enk AH, Mahnke K. 2010. Adenosine triphosphate (ATP) activates regulatory T cells (Treg) in vivo during contact hypersensitivity reactions. J. Immunol. 184:3408-3416. (IF 5,1) Bedke T, Pretsch L, Karakhanova S, Enk AH, Mahnke K. 2010. Endothelial cells augment the suppressive function of CD4+ CD25+ Foxp3+ regulatory T cells: involvement of programmed death-1 and IL-10. J Immunol. 184:5562-70. (IF 5,1) Ring S, Oliver SJ, Cronstein BN, Enk AH, Mahnke K. 2009. CD4(+)CD25(+) regulatory T cells suppress contact hypersensitivity reactions through a CD39, adenosine-dependent mechanism. J Allergy Clin Immunol. 123:1287-96. (IF 10,3) 117 APPENDIX II Areas of research expertise Dendritic cells, Regulatory T cells, Allergy, Tumorimmunology PhD Students Michael Maas. Targeting dendritischer Zellen mittels DEC205-Rezeptor-spezifischer Single chain Fragment variable zur Inhibition von Toleranz sowie zur Induktion von Immunität 20082011. Cum laude Sabrina Schmitt. Funktion und Induktion regulatorischer T-Lymphozyten im murinen Modell der Multiplen Sklerose und während Graft-versus-Host Erkrankung nach Stammzelltherapie. 20072010. Magna cum laude. MD Students Anna Pushkarevskaya. Analysis of CHS reactions in CD73 deficient mice. 2013 ongoing. Current funding DFG Einzelantrag. Bis 3/2014 “Intrazelluläres Targeting des Antigenrezeptors DEC-205“. APPENDIX II 118 PROF. DR. MED. HUGO H. MARTI C3 Professor for Physiology University of Heidelberg Institute of Physiology and Pathophysiology Im Neuenheimer Feld 326 69120 Heidelberg 06221 54 4138 (Fon) 06221 54 4562 (Fax) [email protected] Curriculum vitae since 20032001-2003 2001 1996-2001 1992-1996 1991-1992 1991 1983-1990 C3 Professor and leader of AG Neurovascular Physiology, Institute of Physiology and Pathophysiology, University of Heidelberg, Germany Group leader, Institute of Physiology, University of Zürich, Switzerland Venia legend for Physiology (University of Giessen) Research Associate, Max-Planck-Institute for Physiological and Clinical Research (Prof. W. Risau), Bad Nauheim, Germany Postdoctoral Associate, Institute of Physiology (Prof. C. Bauer), University of Zürich Postgraduate Course in Experimental Medicine and Biology, University of Zürich Promotion to Dr. med. (University of Zürich) Medical College, University of Zürich, Switzerland Areas of research expertise Hypoxia and Ischemia, Neurovascular Biology, Blood-Brain Barrier Schoch HJ, Fischer S, Marti HH. 2002. Hypoxia-induced vascular endothelial growth factor expression causes vascular leakage in the brain. Brain 125: 2549-2557. (IF 9,92) Wang Y, Kilic E, Kilic U, Weber B, Bassetti C, Marti HH*, Hermann DM* (*equal contribution). 2005. VEGF overexpression induces post-ischemic neuroprotection, but facilitates hemodynamic steal phenomena. Brain 128: 52-62. (IF 9,92) Cvetanovic M, Patel J, Marti HH, Kini AR, Opal P. 2011. VEGF ameliorates the ataxic phenotype in spinocerebellar ataxia type 1 (SCA) mice. Nature Med 17:1445-7. (IF 22,86) Kunze R, Zhou W, Veltkamp R, Wielockx B, Breier G, Marti HH. 2012. Neuron-specific prolyl-4hydroxylase domain 2 knockout reduces brain injury after transient cerebral ischemia. Stroke 43:2748-56. (IF 6,16) 119 APPENDIX II 5 selected (most important) publications Marti HH, Risau W. 1998. Systemic hypoxia changes the organ-specific distribution of vascular endothelial growth factor and its receptors. Proc Natl Acad Sci USA 95: 15809-15914. (IF 9,74) PhD/MD students (last 5 years) and titles of their theses Philipp Barteczek, Einfluss der neurone-spezifischen FIH-1-Defizienz bei zerebraler Ischämie, MD, 2014-2015 Jieming Lin, Einfluss von Fumarsäure-Estern auf das Verhalten von Neuronen unter ischämischen Bedingungen, MD, 2013-2014 Daniel Gruneberg, Einfluss der neuronen-spezifischen PHD2-Defizienz auf das räumliche Gedächtnis bei zerebraler Oligämie, MD, 2013-2014, Li Lexiao, Role of the PHD-FIH-HIF axis for the endogenous adaptive response against ischemic stroke, PhD, 2013 – 2016, Reischl Stefan, Effekte der PHD-Inhibition auf die Integrität der Blut-Hirn-Schranke unter ischämischen Bedingnungen, MD; 2012-2014 Bauer Alexander, Molekulare Mechanismen hypoxie-induzierter Permeabilitätserhöhung an der BlutHirn-Schranke, Dr. sc. hum., 2005-2010, magna cum laude Springmann Georg, Inhibierung der hypoxiebedingten Permeabilitätserhöhung in zerebralen Gefäßen durch Blockade des VEGF /VEGF-Rezeptor Systems, MD, 2007-2010, cum laude Mühlhofer Wolfgang; Transienter neuroprotektiver Effekt von B-Vitaminen bei experimenteller Epilepsie im Mausmodell, MD, 2007-2010, magna cum laude, together with Prof. J. Schenkel Staub Janina, Effekte einer permanenten unilateralen Ligatur der arteria carotis communis auf cognitive Verhaltensparameter sowie Proteinexpression und Apoptose im Gehirn VEGF-transgener C57Bl/6 Mäuse, MD, 2006- 2010, magna cum laude, together with Prof. K. Plaschke Current extramural funding Die Rolle von PHD2 für die Regeneration nach Schlaganfall, 2013-2015, Else Kröner Fresenius Stiftung (Kunze/Marti) APPENDIX II Protektion vor ischämischem Schlaganfall durch Hemmung der Prolyl-4-Hydroxylasen, 2013-2015, B. Braun-Stiftung 120 DR. MARTIN RONALD SPRICK Junior Group Leader at HI-STEM gGmbH (Heidelberg Institute for Stem Cell Technology and Experimental Medicine) at the DKFZ Heidelberg, Germany Phone: +49-6221-423913 Fax: +49-6221-423902 E-mail: [email protected] 2006-2009 EMBO Postdoctoral Fellow Laboratory for Experimental Oncology and Radiobiology, AMC UvA, Amsterdam, The Netherlands. (Head of Group: Prof. Jan Paul Medema) 2004-2006 Postdoctoral Fellow Group for Apoptosis Regulation, DKFZ Heidelberg, Germany (Head of Group: Dr. Henning Walczak) 1999-2004 PhD Student and PhD Thesis „Biochemical analysis of the TRAIL-death inducing signalling complex“. Summa cum Laude . DKFZ Heidelberg, Germany and University of Konstanz, Germany. Supervision by Prof. Peter Krammer and Dr. Henning Walczak 1998-1999 Diploma Study and thesis Differential Signal-Transduction and Gene-Expression in Ha-Ras Transformed LiverEpithelial Cells”. Grade 1.0. University of Konstanz, Germany. Supervision by Prof. Ernesto Bade Areas of research expertise Experimental Oncology, Cancer Stem Cells, Cell Death, Cancer Models 5 selected (most important) publications (Impact Factor 2012) Hofner T, Macher-Goeppinger S, Klein C, Rigo-Watermeier T, Eisen C, Pahernik S, Hohenfellner M, Trumpp A, Sprick, M.R. 2013. Development and characteristics of preclinical experimental models for the research of rare neuroendocrine bladder cancer. J Urol. 190(6):2263-70. (IF 3.67) Borovski, T., P. Beke, O. van Tellingen, H. M. Rodermond, J. J. Verhoeff, V. Lascano, J. B. Daalhuisen, J. P. Medema and M. R. Sprick. 2012. Therapy-resistant tumor microvascular endothelial cells contribute to treatment failure in glioblastoma multiforme. Oncogene. doi:10.1038/onc.2012.172. (IF 7.38) Sprick, M. R., M. A. Weigand, E. Rieser, C. T. Rauch, P. Juo, J. Blenis, P. H. Krammer and H. Walczak. 2000. FADD/MORT1 and caspase-8 are recruited to TRAIL receptors 1 and 2 and are essential for apoptosis mediated by TRAIL receptor 2. Immunity 12(6): 599-609. IF (19.80) Vermeulen, L., E. M. F. De Sousa, M. van der Heijden, K. Cameron, J. H. de Jong, T. Borovski, J. B. Tuynman, M. Todaro, C. Merz, H. Rodermond, M. R. Sprick, K. Kemper, D. J. Richel, G. Stassi and J. P. Medema. 2010. Wnt activity defines colon cancer stem cells and is regulated by the microenvironment. Nat Cell Biol 12(5): 468-476. (IF 20.76) 121 APPENDIX II Curriculum vitae 2009-present Junior Group Leader at HI-STEM GmbH (Heidelberg Institute for Stem Cell Technology and Experimental Medicine), Vermeulen, L., M. Todaro, F. de Sousa Mello, M. R. Sprick, K. Kemper, M. Perez Alea, D. J. Richel, G. Stassi and J. P. Medema. 2008. "Single-cell cloning of colon cancer stem cells reveals a multi-lineage differentiation capacity." Proc Natl Acad Sci U S A 105(36): 13427-13432. (IF 9.74) PhD/MD students (last 5 years) and titles of their theses PhD Borovski, Tijana; Cancer Stem Cell Niche: The Place to Be. 2007-2012. Co-promoter. Center of Experimental and MolecularMedicine of the Academic Medical Center (AMC) in Amsterdam, The Netherlands. 2012, Anja Schillert: “Identification and functional analysis of slowly cycling cells in colorectal cancer” (Co-Supervision with Prof. Trumpp) 2012, Christian Eisen: “Development and investigation of a novel model system representing all three subtypes of pancreatic ductal adenocarcinoma revelas novel biomarkers and distinct drug sensitivities” (Co-Supervision with Prof. Trumpp) 2013, Steve Wagner ”Identification of Tumor Initiating Cells in a Patient-Matched Model of Serous Ovarian Carcinoma” (Co-Supervision with Prof. Trumpp) APPENDIX II 122 Appendix III APPENDIX III Declarations regarding Section 9.2 “Collaboration with other Cooperation Partners” 123 124 125 126 127 128 Deutsche Forschungsgemeinschaft Herrn Dr. Anselm Fremmer Kennedyallee 40 D-53175 BONN Germany December 18, 2013 Research Training Group „Hallmarks of Skin Cancer“ Dear Dr. Fremmer, On behalf of King’s Health Partners and the St. John’s Institute of Dermatology, I am delighted to commit faculty and facilities to support the Research Training Group “Hallmarks of Skin Cancer”. We are extremely excited by the potential for innovative developments in this highly important, but under-studied area. Prof. Goerdt and his colleagues have assembled an outstanding set of investigators across the Mannheim-Heidelberg faculties and the German Cancer Research Centre. We commit to supporting their efforts by pairing investigators with outlined projects creating opportunities for intense scientific collaboration, consultation and trainee mobility to work in the laboratories of the members of the London faculty. The London faculty of the RTG provides thorough expertise across a spectrum of relevant basic science and clinical practise. We shall also work together to establish a regular scientific exchange with our German collaborators and the students by co-organizing yearly British-German Workshops on Skin Cancer Biology that will mutually be held at the St. John’s Institute or at the Institutes of the Heidelberg-Mannheim Skin Cancer Alliance. As you may be aware, the St John’s Institute has a world-leading reputation for translational research excellence that goes back many decades. Thus, it can be immensely valuable for the students in the training programme to learn how the Institute has achieved its goals in this critically important arena. We look forward to an excellent interaction that can at the same time promote scientific and clinical progress in skin cancer as well as the development of a new cadre of researchers in this key biomedical area. Sincerely, Adrian Hayday, PhD, FMedSci. Kay Glendinning Professor of Immunobiology, King’s College London School of Medicine. 129 Co-Lead Clinical Academic Grouping in Genetics, Rheumatology, Infection, Immunity & Dermatology, King’s Health Partners. Guy’s Hospital, London SE1 9RT., UK Senior Group Leader, London Research Institute, Cancer Research UK. London, WC2A 3LY, UK 130