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Annual Rep ort 2006
Department of Medical Oncology
VU University Medical Center
Amsterdam, The Netherlands
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Annual Rep o rt 2006
Department of Medical Oncology
VU University Medical Center
Amsterdam, The Netherlands
Editor:
Dr. H.J. Broxterman
Department of Medical Oncology
VU University Medical Center
De Boelelaan 1117
1081 HV Amsterdam
The Netherlands
www.vu-medicaloncology.org
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Contents
C ONTENT S
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DEPARTMENT STAFF
4
INTRODUCTION
8
SYMPOSIA, LECTURES AND COURSES
9
PHASE I/II CLINICAL RESEARCH
12
PSYCHO-ONCOLOGY AND PALLIATIVE CARE SECTION
14
ONCOLOGY WARD 17
OUTPATIENT CLINIC AND DAY CARE UNIT
19
ACTIVITIES OF THE RESEARCH DIVISIONS
Division of Angiogenesis
Division of Gene Therapy
Division of Immunotherapy
Division of Pharmacology
Oncoproteomics Laboratory 20
20
24
28
34
39
PhD THESES
43
SCIENTIFIC PUBLICATIONS
44
C OLO F ON
Copyright © 2007, Department of Medical Oncology, VU University
Medical Center, Amsterdam, the Netherlands. All rights reserved. No part
of this publication may be reproduced or transmitted in any form or by
means, electronic or mechanical, including photocopying, recording, or any
information storage and retrieval system, without the written permission
of the Department of Medical Oncology, VU University Medical Center.
Production: Lobbezoo Medical Communications, Hilversum, the
Netherlands.
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Department staff
D
C l i n i ca l S t aff
Prof. G. Giaccone, MD, PhD, head of department
Prof. E. Boven, MD, PhD
Dr. J. Buter, MD, PhD
Dr. A.J.M. van den Eertwegh, MD, PhD
Dr. W.R. Gerritsen, MD, PhD
Dr. C.J. van Groeningen, MD, PhD
Dr. K. Hoekman, MD, PhD
Dr. B. Kuenen, MD, PhD
Prof. H.M. Pinedo, MD, PhD
e p ar t m e n t
C l i n i ca l S t aff
T ra i n i n g
s t aff
in
Drs. A. Beeker, MD (until May 1)
Drs. H.P. van den Berg, MD
Drs. A. Goosens, MD (until March 1)
Drs. S. van den Heiligenberg, MD
Drs. A. Keijzer, MD
Dr. J.J. van der Vliet, MD, PhD
Psycho-Oncology and
P a l l i at i v e C ar e S t aff
M.S.A. Boddaert, MD, palliative care physician
Dr. M.H.M. van der Linden, PhD, clinical
psychologist
S u p p o r t i v e S t aff
Members of clinical, laboratory and nursing staff during
a brain-storm session. Standing from left to right:
dr. C. Jiménez, dr. A.J.M. van den Eertwegh,
dr. F.A.E. Kruyt, dr.B. Kuenen, dr. H.J. Broxterman,
prof. J. Lankelma, dr.V.W. van Beusechem,
dr. K. Hoekman, dr. J. Buter, prof. G.J. Peters,
prof. V.W.M. van Hinsbergh; Sitting from left to right:
H. Peltenbrug, dr. T.D. de Gruijl; In front from left to
right: M.S.A. Boddaert, prof. E. Boven, dr. M.H.M.
van der Linden, dr. W.R.Gerritsen..
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L. Groenewegen, IT consultant
M.R. Regtuit, financial manager
Drs. A. Carper, MD, personal assistant to prof.
H.M. Pinedo
O n c o l o g y R e s e arc h L a b o rat o r i e s
Prof. G.J. Peters, PhD, head of laboratories
Dr. F.A.E. Kruyt, PhD, molecular biologist,
deputy head of laboratory
Division of Angiogenesis
Prof. V.W.M. van Hinsbergh, head of division
Prof. E. Boven, MD, PhD, laboratory head
Dr. K. Hoekman, MD, PhD, project leader
Prof. J. Lankelma, PhD, chemist, project leader
Dr. H.J. Broxterman, PhD, biochemist, project leader
M.N.A. Bijman, MSc, medical biologist (until
September 1)
Dr. W.P.H. de Boer, PhD, theoretical physicist (guest)
Dr. R. Fernández Luque, PhD, physicist (guest)
Dr. R. Fijneman, PhD, molecular biologist
Dr. A. Greijer, PhD, biologist (until February 1)
Dr. M.L. Janmaat, PhD, biologist (until
November 1)
H. van Cruijsen, MD
A.A.M. van der Veldt, MD
Dr. B.C. Kuenen, MD, PhD
Dr. R.R. de Haas, PhD, senior research
technician
Ing. M.P.A. van Berkel, technician
Ing. H. Dekker, technician
Ing. P.M. Delis-van Diemen, technician
Ing. Y. Yuana, Msc, technician (until August 1)
L. Vroling, MSc, medical biologist
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Division of Gene Therapy
Dr. W.R. Gerritsen, MD, PhD, head of division
Dr. V.W. van Beusechem, PhD, medical biologist,
laboratory head
Prof. C. Dirven, MD, neurosurgeon
Ing. A. Huizenga, technician
Dr. F.H.E. Schagen, PhD, biochemist
(until August 1)
Dr. M.L.M. Lamfers, PhD, medical biologist
(until August 1)
S. Idema, MSc, MD
Ing. I.H. van der Meulen-Muileman, technician
Ing. S. Moeniralm, technician
Division
Immunotherapy (in
c o o p e r at i o n w i t h t h e D e pa r t m e n t o f
P at h o l o g y )
Dr. A.J.M. van den Eertwegh, MD, PhD,
oncologist/immunologist, head of division
Prof. R.J. Scheper, PhD, tumor immunologist,
laboratory head
Dr. T.D. de Gruijl, PhD, tumor immunologist,
group leader
Dr. H.J. Bontkes, PhD, tumor immmunologist
Dr. B. Hangalapura, PhD biologist (as of May 1)
Dr. D. Oosterhoff, PhD medical biologist
S.J.A.M. Santegoets, MSc, medical biologist
J. Lindenberg, Msc, medical biologist (as of 10
August 1)
J. Molling, MSc, medical biologist
M. Moreno, Msc, medical biologist
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R. van de Ven, MSc, medical biologist
H. van Cruijsen, MD
B.J.R. Sluijter, MD
S.M. Lougheed, MSc, medical biologist,
technician
Ing. N. Ravenhorst, technician
Ing. A.W. Reurs, technician
Ing. A. Stam, technician
P.G.J.T.B. Wijnands, MSc, medical biologist,
technician
D i v i s i o n o f P h a r m ac o l o g y
Prof. G. Giaccone, MD, PhD, head of division
Prof. G.J. Peters, PhD, biochemist, head of
laboratory
Dr. F.A.E. Kruyt, PhD, molecular biologist,
deputy head of laboratory
Dr. J.A. Rodriguez, PhD, molecular biologist,
project leader
I.V. Bijnsdorp, MSc, med. biologist
A. Checinszka, MSc, biochemist (until February 1)
C. Lemos, MSc, biochemist
J. Sigmond, MSc, medical biologist (until April 1)
J. Voortman, MSc, MD
A. Watts, Msc, biologist (as of June 1)
Dr. R.J. Honeywell, PhD, senior research
technician
Ing. K. Smid, senior research technician
Ing. S. Span, senior research technician
A. Adema, MSc, technician
Ing. K. Floor, technician
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Ing. M. Gallegos-Ruiz, Msc
Ing. E. Hoebe, technician
Ing. G.A.M. Kathmann, technician
Ing. A.C. Laan, technician
Ing. P. Noordhuis, MSc (as of October 1)
Ing. E. Torun, technician (until July 1)
O n c o p r o t e o m i c s L a b o r at o r y
Dr. C. Jiménez, PhD, chemist, laboratory head
Dr. J.C. Knol, PhD, chemist (as of April 1)
Dr. S. Piersma, PhD, chemist (as of May 1)
Z. El Filali, MSc, technician (as of May 1)
Dr. T. Pham, PhD, bioinformatics
(as of October 1)
P s y c h o - o n c o l o g y a n d P a l l i at i v e
Care Section
Dr. W.R. Gerritsen, MD, PhD, chairman
M.S.A. Boddaert, MD
E.J. Douwes Jr., lay expert
F. Eskens, medical social worker,
Department of Medical Social Work
Dr. K. Hoekman, MD, PhD
Dr. M.H.M. van der Linden, PhD,
clinical psychologist
J. van Ooijen, nurse oncology day care unit
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O n c o l o g y W ar d
N u rs i n g S t aff
R e s e arc h N u rs e s
H. Peltenburg, head oncology ward & outpatient
clinic
C. Stouthart, deputy head nurse
T. Blankwater
A. Bos
H. Bos
J. Brandenburgh
A. Dekkers
M. Douwes
A. Frantzen
A. Hellebrekers-Bos Eijssen
L. Jansma
B. Jorritsma
S. Kooiman
J. van de Lans
J. Oosterling
E. Pauw
H. Refat
T. Rensink
R. Schopenhauer
J. Schouten
J. van Staveren
A. Stapert
L. Tuinman
J. ter Veen
B. Verdegaal
M. IJmker-Westeneng
H.E. Gall
E. Doeleman
I. van der Horst
R. Ruijter
C. Tillier
W ar d A ss i s t a n t s
D. van Giffen
C. Hogers
C. de Reus
M. Verhoef
T. van Os
A d m i n i s t rat i v e S t aff
O n c o l o g y W ar d
I. Charan
B. Guman
M. van den Haak
J. Commandeur
M.L.C. Latupeirissa
M. Secici
D at a M a n a g e rs
K. Groot
V. Hartog
F. Terpoorten
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N u rs i n g S t aff
D ay C ar e U n i t
N. Boere
K. van der Jagt
T. Graas
I. Kamping-Steenkist
B. Krolis
M. Koeree
L. Lenten-Cetinel
J. van Loo
J. van Ooijen
A. Paulus
M. Piet-Krijnen
C. Vree-Bergh
M. Nunez Sandez, departmental assistant
A d m i n i s t rat i v e S t aff
O u t pat i e n t C l i n i c
J. Stolker, head secretary
S. van der Feen
J. van der Heide
A. Kuipers
I. de Ruyter
L. van Straten
J. Versteeg
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V o l u n t e e rs
A. Bisterbosch
J. Lantink
C. Manshanden
H. de Winter
S e cr e t ar i a l S t aff
M.H. Plieger, head secretary
C. Clarinda-Overmeer
S. van Geloven
M. Klaassen
P. Voogd
Secretarial and support staff. From left to right:
S. van Geloven, P. Voogd, M. Regtuit, M. Klaassen,
C. Clarinda-Overmeer. Front: M.H. Plieger.
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In t r o d u c t i o n
I
The year 2006 has seen an increase in the
number of clinical studies started by the
department as well as in the number of
patients included in clinical studies. A strong
program focussing on phase I-II studies and on
translational studies has been firmly established.
In general, a shift towards targeted therapies and
molecular research related to the mechanisms of
action of those therapies has been seen. In fact,
within the phase I unit emphasis has been on the
development of agents targeting the Epidermal
Growth Factor (EGF) receptor, angiogenesis
inhibitors, as well as novel immunotherapies.
A major development in 2006 was moving the
laboratory facilities of the department into the
new building of the Cancer Center Amsterdam
(CCA), located adjacent to the outpatient clinic
of the hospital. CCA opened its doors officially
on June 19, 2006. Also, the new core facility for
proteomics , established in collaboration with
other departments interested in using this novel
technology platform, got its place in the new
CCA building. The new CCA laboratory facility
aims at bringing together under one roof all
research laboratories of the major departments
of the VU University Medical Center conducting
cancer research (i.e., medical oncology, pathology,
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haematology, head and neck surgery, dermatology
and pediatric oncology).
New cancer research laboratory of the Cancer Center
Amsterdam
up a student exchange program with both the
University of Bologna, Italy, and the University of
Turin, Italy, with students from both universities
spending a number of months in our laboratory
to gain research experience.
A selection from the diversity of activities of
the Department of Medical Oncology has been
highlighted here. These and other activities are
described in more detail in this Annual Report.
We are confident that ongoing activities, new
developments and initiatives in our research,
organization (phase I/II unit) and education will
lead to further improvements in cancer patient
care in the years to come.
In the year 2006, our international collaboration
with foreign institutions has continued, in
particular with Johns Hopkins Cancer Center
in Baltimore (HIF-1 research), Institut GustaveRoussy in Paris (gene therapy of malignant
gliomas), University of Alabama in Birmingham,
Alabama (novel viral vectors for gene therapy),
and Harvard University (angiogenesis). Our
collaboration with Johns Hopkins is fostered
by an international grant from Aegon. Prof. Van
Hinsbergh coordinates this line of research for
the department. The department has also set
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Symposia, lectures and courses
S
y m p o s i a
TAT 2 0 0 6, M arc h 1 6 - 1 8 ,
Amste rdam ,
The Netherlands
In March 2006, the 4th International Symposium
on Targeted Anticancer Therapies (TAT 2006) was
held at the VU University in Amsterdam. This
meeting was organised by the NDDO Research
Foundation (congress director: Dr. M.W.
Lobbezoo; www.nddo.org ) together with the
European Society for Medical Oncology (www.
esmo.org) and the Department of Medical
Oncology. The department head, prof. G.
Giaccone, served as Symposium President. The
TAT series builds on a tradition of symposia
organised at the VU on new anticancer drug
development. TAT 2006 demonstrated that
this annual meeting series has come of age. It
provides an excellent forum for the presentation
and discussion of virtually all aspects of targeted
anticancer therapies in (pre)clinical development.
Compared to the meeting in 2005, some
important adjustments in the focus and the
format of the meeting have been implemented
successfully in 2006. These included the
focussing on early-phase clinical development,
,
l e c t u r e s
a n d
resulting in more original presentations on phase
I and II studies, and stricter abstract review,
resulting in a higher rejection rate and better
overall quality of the scientific presentations.
Overall, well over 400 people representing 42
different countries attended TAT 2006, a clear
increase in attendance over TAT 2005. About
70% of delegates were from European countries,
20% from the USA and Canada, and 10% from
the rest of the world. Compared to TAT 2005,
the relative size of the delegations from Asia and
the rest of the world has increased at the expense
of European delegations, illustrating the further
globalization of the TAT series. Delegates’
evaluations of various aspects of TAT 2006
ranged between 3.2 and 4.2 on a scale from 1 to 5.
TAT 2006 witnessed the start of a special task
force on ‘Methodology for the Development of
Innovative Cancer Therapies’ (MDICT). This
forum of experts from leading academic cancer
research institutes, supplement with observers
from, industry and regulatory agencies, attempts
to develop practical guidance on the optimal
development of innovative anticancer agents.
Dr. Elizabeth Eisenhauer presented an extensive
summary of MDICT’s first session on important
issues in the design of phase I studies of targeted
c o u rs e s
agents in the main symposium. A separate report
on the first MDICT meeting has been offered
for publication in a scientific journal.
One of the highlights was the NDDO Award
Lecture on ‘Adventures in targeted cancer
therapy: From tumour hypoxia to the cancer
genome’, presented by professor Paul Workman
from the Institute of Cancer Research in Sutton,
UK. Professor Workman and his co-workers have
an impressive track record in drug design and
discovery as well as in early-phase clinical and
translational research.
Professor Paul Workman (left) receives the NDDO
Award 2006 from Dr. Coenraad van Kalken, Director
NDDO Research Foundation (right) during the TAT
2006 Opening Ceremony.
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Much attention has recently been drawn by
drugs showing promising clinical activity in
advanced renal cell cancer. In his Keynote
Lecture, professor Martin Gore reviewed the
most advanced drugs in clinical development,
sorafenib (Nexavar®) and sunitinib (Sutent®),
together with several promising drug candidates
with less mature data. Sorafenib and sunitinib
have recently obtained regulatory approval for
this indication in the USA, the European Union,
and several other countries.
Among the many agents in active preclinical
and clinical development and discussed during
TAT 2006, AMN107 stood out by its remarkable
clinical activity in a phase I study in imatinibresistant CML. AMN107 is a novel inhibitor
of Bcr-Abl, the target of imatinib (Glivec®) in
chronic myeloid leukaemia.
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O p e n d ay f o r pat i e n t s :
“ K a n k e r a n d e rs b e l i c h t ”
M ay 3 , 2 0 0 6
On May 3, 2006, we celebrated the fifth
anniversary of the VU University Medical Center,
which is a combination of the former VU Hospital
and the VU School of Medicine. For our patients
we organized a memorable program “Kanker
anders belicht”, (“a different light on cancer”),
loaded with live music and interesting information
for cancer patients. Mary-Lou van Steenis
elegantly presented the program. Among others,
she introduced the choir “Zingen voor je leven”
(“Singing for life”) directed by Harry de Beer .
The choir makes use of different songs to express
their emotions related to (previous) illness, such
as “This little light of mine”, “What a wonderful
world” and “With a little help from my friends”.
The next meetings, TAT 2007 and TAT 2008,
have been scheduled for March 8-10, 2007 and
March 20-22, 2008, respectively.
The choir “Zingen voor je leven” directed by
Harry de Beer
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Harry de Beer
Jacobine Donker being interviewed by
Mary-Lou van Steenis
Mr. Frits J. Duparc, MSc, director of the
Mauritshuis Museum, The Hague, showed
old paintings expressing loyalty and care. True
friendship, fidelity in marriage, and care for the
sick have often been subjects of imaginations on
canvas. Jacobine Donker directed a short movie
in which she followed two patients in their daily
life. Both have to cope with limitations caused
by their treatment, but can live a happy life. A
short piano intermezzo was given by Mark van
der Feen. Prof. Paul van der Valk demonstrated
the possibilities the modern pathologist has to
discover tumor characteristics at the molecular
level. Laboratory research aimed at improving
cancer treatment was illustrated by a short movie
directed by Andrew Fallon. He followed a patient
who participated in a clinical immunotherapy
trial and showed the laboratory activities required
in preparation of his treatment. Prof. Giuseppe
Giaccone demonstrated the possibilities we
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develop in the clinic to further improve therapies
directed against specific targets in tumors.
Participants were very enthusiastic about the
market place in the Amstelzaal Foyer, which could
be visited at lunch time. Many organizations were
present, either with focus on a particular tumor
type, on handling symptoms of disease or on
special clothing.
Attentive public in the Amstelzaal
A few written reactions from patients, apart
from the many words of thanks:
- ‘we had a wonderful day and learned a lot’
-‘a very impressive and emotional program,
which enabled us to manage disease in a better
way’
-‘fantastic atmosphere and an excellent day;
why is the next one planned only in 5 years?’
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T e ac h i n g
The department contributed to teaching, both
to the new curriculum for medical students and
to the Master in Oncology program. The Master
in Oncology program was established in 2002
and was recognized by the Minister of Education
as one of two top-masters of the VU University.
In 2006, the master program performed a self
evaluation in order to renew its recognition. Prof.
Peters is chair of the examination committee and
one of the organizers of the course on Innovative
Therapies, in which students are taught on both
standard and novel therapies. This also includes
an intensive study of the literature to formulate,
present and defend a research project. In 2006,
twenty-five students passed their master. Some
of the new masters in oncology continued their
career by doing a Ph.D. thesis in the Department
of Medical Oncology.
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Phase 1/11 clinical research
P
O r g a n i z at i o n
h as e
and
fac i l i t i e s
As in 2004 and 2005, the phase I/II research
unit has been used very intensively in 2006.
The unit consists of a three-bed patient room
on the oncology ward with beds equipped with
2 monitoring devices, offering the possibility
to monitor heart rate, blood pressure, oxygen
saturation and body temperature of patients
participating in trials. With the devices, also
EKGs and analyses of heart rhythm (72 hour
full disclosure) can be performed. In 2005, the
laboratory for processing and storage of blood
samples moved to a larger room near the phase
I/II unit. It is equipped with centrifuges, flow
chamber, and -20°C and -80°C freezers with a 24
hour temperature alarm system.
The aim of the research unit is to improve care
for patients participating in trials and professional
competence in adhering to study protocols. Due
to the increasing complexity of drug development
in recent years, a dedicated team is required.
The research unit is currently managed by five
research nurses: Rita Ruijter, Helen Gall, Inge van
der Horst, Corinne Tillier and Ellen Doeleman.
The unit is supervised by Dr. B. Kuenen, medical
oncologist, who also chairs all the phase I/II
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c l i n i ca l
r e s e arc h
meetings at the department. The phase I/II team
covers the full period from protocol development
to completion of the trials. Their tasks consist
of starting up studies, co-ordination of all trial
related procedures, extra checks of in- and
exclusion criteria, final enrolment of patients,
pharmacokinetic sampling and work-up, planning
visits and assessments according to the protocol,
follow-up of patients, including scoring of toxicity,
all logistics, sample shipment, etc.
Staff of the phase I/II research unit (from left to right):
Ellen Doeleman, Inge van der Horst, Rita Ruijter,
Corinne Tillier, Bart Kuenen, Helen Gall
C l i n i ca l
studies
Pharmacokinetic/pharmacodynamic evaluations
were performed in all phase I studies and some
of the phase II/III studies. A major focus of our
department is, besides participating in phase
II and III trials, performing phase I trials.
The various phase I, II and III studies can be
subdivided into three main categories: those
investigating classical chemotherapy compounds,
those investigating targeted compounds and
those investigating combinations of targeted
compounds or combinations of a targeted
compound and chemotherapy. The targeted
compounds under study in our department are
mainly directed at two targets: the epidermal
growth factor receptor (EGFR) pathway, which
is involved in tumor growth in several tumor
types, and the vascular endothelial growth factor
(VEGF) pathway, which is the main player in the
formation of new blood vessels (angiogenesis).
Examples of trials with EGFR inhibitors are
• a phase I dose escalation study of the fully
humanised monoclonal antibody IMC-11F8;
• a phase I/II study of erlotinib in combination
with radiotherapy in rectal cancer;
• a feasibility phase I/II study of erlotinib added
to radiotherapy plus gemcitabine in locally
advanced irresectable pancreatic cancer;
• a metabolism study of erlotinib in smokers
versus non-smokers;
• a nationwide Dutch trial randomising
patients with colorectal cancer to
capecitabine, oxaliplatin and bevacizumab
with or without cetuximab (CAIRO2)
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Angiogenesis inhibitors under study in our
department include sunitinib, SU014813,
AZD2171, BMS582664, VEGF-trap, and
bevacizumab. Other targets investigated in the
clinic are the apoptotic machinery (anti-survivin
and AMG951), which is also extensively studied
in our preclinical department, and m-TOR
(RAD001). Another main focus of our clinical
research program is immunotherapy with a
special interest in vaccinations combined with an
anti-CTLA4 monoclonal antibody (MDX010)
or chemotherapy. A number of trials with these
combinations are open for accrual in prostate
cancer and melanoma.
The department aims at performing translational
research and therefore many side-studies
related to the clinical trial protocols are being
performed. Examples are an extensive analysis
of the EGFR expression patterns, mutational
status and downstream effects in tumors of
patients treated with EGFR-inhibitors, analysis
of maturation of dendritic cells during treatment
with angiogenesis inhibitors, the mechanism
of hypertension in patients treated with
angiogenesis inhibitors, detection of circulating
endothelial cells and precursors in patients
treated with angiogenesis inhibitors, extensive
immunological monitoring of patients in the
vaccination/anti-CTLA4 trials and performing
proteomics of blood samples.
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Psycho-oncology
and palliative care section
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s y c h o
O r g a n i z at i o n
o n c o l o g y
The section of Psycho-oncology and Palliative
Care continued its efforts to improve the quality
of care within the department*. In 2005, the
section started a scientific research network in
collaboration with the Department of Medical
Psychology, the Department of Otolaryngology,
Head and Neck Surgery, the Department of
Social Medicine and the Ingeborg Douwes
Center for Psycho-Oncology to investigate
quality of life and palliative care. The focus in
2006 was on developing new scientific projects.
This led to grant applications for several
projects concerning an individualised quality of
life intervention for hospitalised patients with
advanced cancer, the selection and validation of
instruments for measuring outcome of care in
oncology patients without curative treatment
options and screening of ‘high risk’ patients
with the Emotional Distress Thermometer (J.
Holland, 2005). We plan starting these projects
in 2007.
In 2006, psychological and palliative care has
been extended to patients of several departments
in the VU University Medical Center. Besides
providing patient care, both dr.M.H.M. van der
Linden and drs. M.S.A Boddaert, continued
educating health practitioners in psychosocial
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a n d
pa l l i a t i v e
and palliative care to enhance professional
expertise in the field of care for cancer patients.
In addition, both the medical faculty and the
hospital education centre extended psychooncology and palliative care education in their
curriculum for medical students and oncology
nurses.
In collaboration with the regional Comprehensive
Cancer Center Amsterdam (IKA) and Leiden
University Medical Center, master classes on
psycho-oncology for medical students and
psychology students were organized.
P a l l i at i v e
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s e c t i o n
double rooms on the ward were transformed
to single rooms and fitted with rooming-in
capacity**. Official opening of these rooms took
place in October 2005 and 115 patients were
admitted to these rooms for symptom control
throughout 2006.
car e
A comprehensive business plan to provide
multidisciplinary palliative care to the
Department of Medical Oncology gained
hospital-wide support and was implemented in
2004 and 2005. The Board of Directors of the
VU Univeristy Medical Center acknowledged
palliative care as an important area of expertise
to be developed within the center and therefore,
the implementation of our plans took place in
close collaboration with all departments involved
in setting up the VUmc Expertise Centre for
Palliative Care. As a preliminary result, four fully
operational palliative care beds were opened on
the oncology ward in 2005. For this purpose, four
View of palliative care room on the oncology ward
As a result of increased hospital-wide awareness
of palliative care, referrals to the department’s
consultant in palliative medicine increased
steadily to 125 new consultations throughout
2006. Close collaboration within the hospital’s
Palliative & Supportive Care Team between prof.
dr. W.W.A. Zuurmond, MD, anaesthesiologist,
drs. P. Doornaert, MD, radiotherapist, drs.
M.S.A. Boddaert, MD, dr. M.H.M. van der
Linden and B.Tuit, specialist nurse, has led
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to the availibility of 24-hour palliative and
supportive care for patients admitted to our
hospital. In addition, as a courtesy to the regional
Comprehensive Cancer Center Amsterdam
(IKA) the hospital’s Palliative and Supportive
Care Team is available for regional consultation
by phone outside office hours and during
weekends.
Through these interdepartmental collaborations
the Department of Medical Oncology is now an
active participant in the VUmc Expertise Centre
for Palliative Care that was launched in October
2006.
Psycho-oncology
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Oncology hostesses
The department’s oncology hostesses work as
volunteers. In 2006 again they did a great job in
accompanying and caring for 637 out of the 782
new patients of our outpatient clinic.
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The referral system between oncologists and the
department’s clinical psychologist and medical
social worker has been working swiftly and
effectively. The number of referrals is stable:
more than one third of all new patients (329
out of 782 new patients in 2006) consulted
the psychosocial team (medical social worker,
clinical psychologist and consultant in palliative
medicine). In conformity with the ‘National
Program of KWF Kankerbestrijding 2005-2010’,
the psycho-oncology team started introducing a
validated screening instrument to improve the
referral system by which high-risk patients will
be identified.
Dr. M.H.M. van der Linden started a
collaboration with dr. I. Verdonck-de Leeuw
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(psychologist of the Department of
Otolaryngology, Head and Neck Surgery) to
investigate distress in head and neck cancer
patients and their spouses. In 2006, a study was
completed in which group therapy was offered to
these patients and their spouses in collaboration
with the Ingeborg Douwes Center.
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S p e c i a l n u r s e c o n s u ltat i o n
In the outpatient clinic, specialist nurses
continued to provide follow-up care by phone
for patients starting a new line of therapy. The
frequency of weekly phone-in hours for patients
and their families was increased to a daily
frequency, such that they may contact a specialist
nurse about specific problems or situations at
hand.
T r a n s m u r a l P a l l i at i v e C a r e G u i d e lines
Supported by a grant from AGIS healthcare
insurance company, eight transmural Palliative
Care Guidelines for a variety of symptoms (bowel
obstruction, pain, dyspnoea, confusion, nausea
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and vomiting, ascites) were compiled by one of
the department’s senior nurses and the specialist
nurse on the Palliative and Supportive Care
Team. The aim of these guidelines is to inform
healthcare professionals coming to the homes of
our patients on the care that was initiated during
admission. By doing so, we aim at improving
the continuity of care and possibly prevent readmission.
F a m i ly S u p p o r t P r o j e c t
The Family Support Project was initiated in 2001
by prof.dr. H.M. Pinedo. It aims at improving
communication between cancer patients, their
family members and their doctors. Following
tremendous fundraising activities, the first
“Time-Out” family room on the oncology ward
was opened on March 24, 2005***. Here, family
members and patients have the opportunity
to take a break. It is a quiet and comfortable
place in close vicinity to their loved ones, where
an experienced oncology nurse is available for
support and for relieving concerns. So far, the
Family Support Project has been a great succes.
Between March 2005 and December 2006,
300 families used thr ‘Time-Out’ familiy room
to celebrate birthdays and marriages, to do
homework and share joyous and sad moments
together in private.
In 2006, the “TimeOutkamers VU Medisch
Centrum” Foundation initiated new efforts to
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finance family rooms in the outpatient clinic, the
intensive care unit and on the surgical ward to
expand support for the family of cancer patients
throughout the center. The official opening of
the second and third family room is planned to
take place in 2007.
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lso a note of thanks is due to the Ingeborg
A
Douwes Stichting for organizing a benefit
dinner and theatre show at “Theater Carre” in
June 2006, to financially support psychosocial
care of patients with cancer.
** A special note of thanks is due to dr. K.
Hoekman and the community of Urk, the
Roparun Foundation, several individual
sponsors, the Ingeborg Douwes Stichting,
the Telegraaf Puzzelactie and individuals who
joined a benefit dinner at the Mauritshuis
Museum in April 2005 for their generous
financial support.
View of the “Time-Out” family room on the oncology
ward
S pecial notes of thanks
* A special note of thanks is due to prof. H.M.
Pinedo and dr. W.R. Gerritsen of the VUmc
Cancer Center Amsterdam. Their continued
efforts in obtaining financial support for
palliative medicine and psychosocial care have
enabled drs. M.S.A. Boddaert and dr. M.H.M.
van der Linden to continue their work within
the department as well as hospital-wide.
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*** A special note of thanks is due to the
‘TimeOutkamers VU Medisch Centrum’
Foundation for recruiting sponsors for the
Family Support Project. Through its generous
donation to the VUmc Cancer Center
Amsterdam the Department of Medical
Oncology will be able to realise two additonal
family rooms.
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O n c o l o g y Wa r d
O
A number of innovations aimed at increasing
the quality of patient care as well as improving
the quality control process on the oncology
ward were started in 2005 and were continued
enthusiastically this year. Among the activities
are the assignment of a nurse specialist providing
specialist support to family only. This means
that one nurse works 20 hours per week as the
coordinating family room nurse, while at the
same time the ward’s team of nurses work on a
rotation of two days each in the family room.
An important development was the opening of
four palliative care rooms on the ward in late
2005, which are now fully operative. Patients
are admitted to these rooms for a maximum of
11 days per admission. During this period, they
receive multidisciplinary treatment for multiple
symptomatic palliative problems. After the
patient’s symptoms have been regulated, the
patient is discharged to the home situation with
specialist home care, or to a palliative unit of
either a nursing or residential home or one of the
hospices in the Netherlands.
The annual training for nurses in the fall of 2006
covered the following subjects: treatment path
for colon cancer and ongoing studies, depression
and coping with cancer, prevention and
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treatment of stomatitis during chemotherapy,
cachexia and nutrition with cancer and
information about cetuximab. The new
informative movie: “Chemotherapy, what is it?”
for patients who will undergo cancer treatment,
was shown. Both the movie and the training
were well received. This special movie is the final
project of Lisette Saveur and Marleen Westeneng
who were trained as oncology nurses. It took a
while before the plan could be carried out, to
get consent, financing and the question whether
to ask professionals or patient-actors, but
once everything was cleared, the director, Bob
Aardewerk, started with a lot of commitment,
effort and enthusiasm. In addition to the Dutch
version several foreign language versions will be
prepared (Turkish, Moroccon-Arabic) and the
final result will be available by May 1, 2007 for
use in other institutions as well.
In November 2006, the day care unit received a
cheque of €30.000 from “Stichting Roparun”,
which enabled the purchase of two Paxman
equipments for cooling the scalp of patients. This
is a highly valuable extension of our possibility to
improve treatment and in particular the quality
of life of certain patients, who will now suffer less
hair loss caused by chemotherapy treatment. As
of early 2007, this treatment will be offered to
patients.
S af e t y
management
Two projects, which are in fact the forerunners
of an integrated safety management system, are
ongoing. The first project concerns the safety
of managing and administering chemotherapy.
Three SOPs (Standard Operating Procedures)
were developed at the end of this project. In
doing so, stepwise checkpoints were introduced
and everyone’s responsibility in the process from
arranging of cystostatics, to administration to
the correct patient at the correct time and in the
correct dosage has been laid down in writing.
The second project concerns the ‘Safe Reporting
of Incidents” (SRI; blamefree reporting) project.
The objective of this project is to achieve a
decentralised safety management system.This
is a project embedded in the national action
plan from the Ministry of Health, Welfare
and Sport in which 24 leading hospitals have
been designated to improve care and efficiency
in health care. Safety is an important part of
this project. The SRI project on the ward is
multidisciplinary in nature. Nurses and doctors
can report (near) errors and (near) accidents in a
very accessible and easy fashion. ‘Incidents’ is a
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neutral, non-accusing term. Within this system,
the barriers in medical and nursing healthcare
procedures are tested and analysed with regard
to their functionality. Every incident reported is
analysed by a multidisciplinary working group,
which meets weekly. Reporting trends are
addressed by means of improvement trajectories,
which will either improve the barriers or place
them earlier in the process such that incidents
either do not reach the patient at all or do not
affect the patient in a significant way.
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Out-patient
clinic and day care unit
u t p a t i e n t c l i n i c a n d d a y car e u n i t
In 2006, 803 new patients were referred to
our outpatient clinic. Thirty-six percent of
our patients are from Amsterdam, 20% from
the province of Noord-Holland outside of
Amsterdam, and the remainder from other
provinces, indicating that a considerable number
of patients were willing to travel (sometimes for
more than an hour) to get access to our care. The
distribution of tumor types among the patients
attending our outpatient clinic and day care unit
is depicted in Figure 1. About one-third of the
patients had a tumor of the digestive tract and
another one-third of the urogenital tract.
Ovarian/cervix
7%
Rest
19%
Lung
6%
ENT
10%
GU tract
18%
Breast
13%
GI tract
27%
Since most of our patients had never visited the
VU University Medical Center before being
referred, all new patients visiting our outpatient
clinic are escorted by a hostess, who will contact
them before travelling to the hospital. They
also explain our procedures to them. After
consultation with a medical oncologist, the
hostess arranges all further appointments for the
patient. The hostess also alerts the patient to the
possibility of counselling by the psycho-oncology
team whenever necessary.
Almost all patients were seen by our staff
within one week of registration. Apart from
patients seeking a second opinion, most patients
preferred to be treated at the VU University
Medical Center. When possible, patients often
consented to participating in one of our clinical
trials. The flexible attitude of all involved in
the day care unit, dealing with complicated
treatments, is very important for the high-quality
care we offer our patients. The continuous
excellent collaboration with the pharmacy
department, blood transfusion service and
clinical laboratories has been instrumental in
achieving this.
Figure 1. Distribution of tumor types among patients
attending the outpatient clinic and day care unit.
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Activities
of the research divisions
c t i v i t i e s o f t h e r e s e arc h d i v i s i o n s
Division
of
Angiogenesis
The Division of Angiogenesis aims at
improving the understanding of tumor-induced
angiogenesis and establishing the clinical value
of anti-angiogenic therapy for cancer patients.
The growth of tumors and their metastases
depends on tumor-induced neo-vascularization
(angiogenesis). The process of angiogenesis
is initiated and maintained by tumor-derived
growth factors, which are generated due to
genetic abnormalities of tumor cells and by local
hypoxia. Hypoxia stabilizes the HIF-1α protein,
which is a transcription factor for many genes
involved in angiogenesis. Among tumor-induced
pro-angiogenic factors, vascular endothelial
growth factor (VEGF) has been recognized as
one of the most important stimulatory factors
of angiogenesis. Consequently, VEGF and its
receptors (mainly VEGFR2 and VEGFR1) are
primary targets for anticancer therapy.
Evidence that inhibition of the VEGFVEGFR pathway is of clinical benefit in cancer
patients is growing. It has been shown that the
monoclonal antibody bevacizumab (Avastin®),
which is directed against VEGF, has activity as
a single agent in renal cell cancer. Moreover, the
20
combination of bevacizumab and chemotherapy
was associated with an increased number of
tumor responses, prolonged time to progression
and prolonged overall survival in colorectal
cancer and non-small cell lung cancer.
Members of the Division of Angiogenesis of Medical
Oncology together with the members of the Department
of Physiology after their weekly common Thursday
morning meeting
In addition, the combination of bevacizumab and
an anti-EGFR agent appeared to be synergistic
in patients with advanced lung cancer. Finally,
several tyrosine kinase (TK) inhibitors of
VEGFR-2 have shown impressive activity in renal
cell cancer (i.e. sunitinib). VEGF is a dominant
factor in the biology of this tumor type. These
encouraging data have led to a sharp increase
in the number of clinical trials evaluating antiVEGF and anti-VEGFR agents with or without
other biological agents, chemotherapy and/or
radiotherapy.
Clinical trials with second- and third-generation
VEGFR and VEGF inhibitors are being
performed. These clinical trials are accompanied
by side studies to learn more about the effects
of anti-angiogenic agents on their target, the
tumor vasculature’s endothelium as well as on
host tissue (e.g. the normal vessel endothelium).
These side-studies include investigations
of circulating endothelial progenitor cells,
immunological parameters, endothelium-specific
marker proteins and complete protein profiles
(proteomics). In addition, effects of antiangiogenic therapy on the normal physiology
of vessels (regulation of the blood pressure) are
being studied and new imaging techniques are
being used to monitor anti-tumor effects.
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The specific research projects of the division are:
• Clinical trials of anti-angiogenic agents
•Circulating endothelial cells (CECs) and
endothelial progenitor cells (CEPs)
• Role of HIF-1 in tumor-induced angiogenesis
• Angiogenesis in ovarian cancer
•Integrative tumor cell biology for
improvement of cancer treatment
Clinical trials of
a n t i - a n g i o g e n i c ag e n t s
In 2006, all data collected in the phase I clinical
trial of SU014813, carried out by prof. dr. E.
Boven in collaboration with the Hamburg
University Hospital, Germany, were evaluated
for toxicity and efficacy. SU014813 is an oral
TK inhibitor of the VEGF receptors 1 and 2,
PDGFRβ, c-Kit and Flt-3. A total of 77 patients
have been included in the trial, 38 at a schedule
of 4 weeks on treatment and one week off, and
39 at a schedule of continuous daily dosing.
Most side effects were, in general, well tolerable.
Few grade 3-4 toxicities occurred and the ones
that did occur were mainly obeserved with the
continuous dose schedule: diarrhea and fatigue
were most prominent. Twelve patients responded
to SU014813, while 32 patients experienced
stabilization of disease. Thirteen patients were
on treatment for a period > 12 months. Presently,
an international phase II trial is ongoing with
SU014813 being administered at 100 mg per day
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continuously in patients with advanced breast
cancer, who were previously treated with a
taxane and an anthracycline. Antitumor activity
has been noticed in several patients, including
patients treated by our department.
An extended access program of SU011248
(sunitinib; Sutent®) in advanced renal cell cancer
was coordinated by prof. dr. E. Boven and dr.
A.J.M. van den Eertwegh and was carried out
by A.A.M. van der Veldt, MSc,) in collaboration
with the group of dr. J. Haanen, Netherlands
Cancer Institute, Amsterdam. A total of 83
patients were treated, including elderly patients
age and patients with poor performance status.
Several patients, especially those with clear cell
carcinoma histology, responded to the drug for
an extended period of time. Of interest, some
patients with their primary tumor still in situ,
known to be resistant to cytokine therapy,
showed massive tumor necrosis upon treatment
with sunitinib. Several side-studies were done
to obtain more insight in the mechanism of
action of sunitinib, in particular a possible role
of circulating endothelial cells and circulating
dendritic cells as well as the mechanism of
hypertension. The data are currently being
analysed.
Presently, a phase I clinical trial of the
combination of sunitinib and irinotecan in
patients with advanced solid malignancies is
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ongoing to determine the doses that can be safely
combined (prof. E. Boven in collaboration with
Institut Gustave Roussy, Paris, France).
H. van Cruijsen, MD (under the supervision
of dr. K. Hoekman and dr. T. de Gruijl) has
performed a phase I/II study with escalating
doses of AZD2171, an oral TK inhibitor of
VEGFR-2, PGDFR-β and c-kit, in combination
with 250 or 500 mg of an oral TK inhibitor of
EGFR (gefitinib). This study is conducted in
cooperation with the Department of Medical
Oncology, University Medical Center Utrecht
and the Department of Medical Oncology,
Radboud University Nijmegen Medical Center.
Side-studies in this trial included dynamic CT
imaging of tumors in a subset of patients and
measuring effects of the therapeutic agents
on circulating dendritic cell subpopulations
(for details of the results, see Division of
Immunotherapy).
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C i r c u l at i n g e n d o t h e l i a l c e l l s
( CEC s ) a n d e n d o t h e l i a l p r o g e n i t o r
c e l l s ( CEP s )
Testing novel anti-angiogenic drug combinations
in the clinic is hampered by the lack of suitable
methods for target validation and efficacy. Strong
evidence from preclinical studies suggests that
the number of circulating endothelial cells
originating from the (tumor) vasculature (CECs)
may serve as an early indicator of anti-angiogenic
and/or antivascular effects. In addition, in
preclinical models, circulating endothelial
progenitor cells (CEPs), which originate from the
bone marrow, have been shown to contribute to
neo-vascularisation. Therefore the quantification
of pre-treatment numbers of CEPs and CECs
and changes in these number during therapy
might be markers of efficacy of anti-angiogenic
therapy.
Currently, there is no validated protocol to
measure these cell populations longitudinally
in cancer patients. Supported by a grant from
the EU-FP6 program “Angiotargeting” (dr.
H.J. Broxterman, dr. K.Hoekman, prof. dr.
V.W.M. van Hinsbergh), we have designed a
flow cytometric method for the quantification
of populations of CECs and CEPs in peripheral
blood of cancer patients in clinical trials.
Since almost all anti-angiogenic agents and
combinations presently under study in the
clinic target VEGF or its receptor VEGFR-2,
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we have in particular focussed on measuring
cell populations expressing VEGFR-2. A study
protocol has been set up to establish the
reliability and reproducibility of a 4-colour
flow cytometry procedure to measure the
frequencies of subpopulations of cells with
endothelial and progenitor markers, including
CD34, AC133, CD146, CD31, and CD105, in
combination with VEGFR-2 as well as CD45 (to
exclude hematopoetic cells). L. Vroling, MSc,
found, when comparing 20 healthy volunteers
and 14 patients with advanced cancer, that
the population of CD45negCD34brightCD133neg
CD146posVEGFR-2pos cells was increased in the
cancer patients. This novel “small EC-like” cell
population is currently being investigated further
in vitro and in clinical trials in patients with
non-small cell lung cancer (NSCLC) treated with
bevacizumab and the EGFR blocker erlotinib
and patients with renal cell cancer treated with
sunitinib for its potential as a biomarker of
anti-angiogenic, antitumor effects. Another
population of circulating cells, previously
identified in a number of studies as CECs, has
been shown to exist mostly of large platelets.
Other studies in colorectal and non-small cell
lung cancer patients are being planned.
Emerging evidence suggests that human blood
contains bone marrow (BM)-derived endothelial
progenitor cells (EPCs) that contribute to
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postnatal neovascularization. Clinical trials
demonstrated that administration of BM-cells
may enhance neovascularization. Most studies,
however, used crude BM cell populations.
Identifying the role of different cell populations
is important for developing improved cellular
therapies. L. Vroling, MSc, and M. Weij have
set up a culture technique for so-called “lateoutgrowth endothelial progenitor cells”, from
human cord blood. These cells are characterised
by a typical cobblestone-like appearance, and
have a marker profile consistent with endothelial
cell lineage (CD45neg CD14neg CD31bright CD146pos
CD144pos). These cells also express EGFR and
will therefore be used to investigate the effects
of combined EGFR and VEGFR inhibition (e.g.
bevazicumab with ertolitinib) on EPCs as models
for tumor endothelial cells.
Role
of
HIF - 1
in tumor-induced
angiogenesis
H ypoxia I nducible F actor -1 (HIF-1)
in
breast and colon tumors
HIF-1 is an important transcription factor that
stimulates tumor growth and metastasis via
several pathways, including the induction of
angiogenesis-regulating growth factors VEGF
and SDF-1. Activation of HIF-1 depends on
the presence of its α-subunit, which in the
presence of oxygen is rapidly degraded after
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proline hydroxylation and subsequent interaction
with von Hippel Lindau (VHL) protein in the
proteasome. Hypoxia increases HIF-1α levels
by inhibiting prolyl-hydroxylase-mediated
hydroxylation and thereby preventing HIF-1α
degradation. In line with such a mechanism
we found that silencing prolyl-hydroxylase-2 in
endothelial cells by siRNA resulted in sustained
activation of HIF-1α.
The expression of HIF-1α-induced genes was
also investigated in biopsies of colon carcinomas,
adenomas and non-malignant colon biopsy
specimens. This was done in collaboration with
prof. G.A. Meijer (Department of Pathology,
VU University Medical Center), supported by
an AEGON fellowship. HIF-1α was clearly
demonstrated in colon tissue and its correlation
with hypoxia, GLUT-1, CAIX and other proteins
was further investigated. Special emphasis
was given to tumor vascularization and the
occurrence of genetic instability in the patients’
biopsies. A mouse colon tumor model was set up
to investigate the role of angiogenesis, hypoxiaassociated factors (including HIF-1α) and
monocytes in the progression of colon tumor (dr.
R. Fijneman).
E xperimental studies on
hypoxia and HIF-1α
In collaboration with dr. M.A. Engelse
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(Department of Physiology, VU University
Medical Center), in vitro parameters of
angiogenesis were studied for prolonged periods
under various oxygen concentrations. mRNA
expression profiles of human macro- and
microvascular endothelial cells kept at hypoxic,
normoxic and ambient air atmosphere for
several months were analyzed. A selection of
the differentially expressed genes is currently
investigated with respect to their possible role in
angiogenesis.
A n g i o g e n e s i s i n o va r i a n c a n c e r In a project carried out by M. Bijman, MSc,
the anti-angiogenic properties of microtubuletargeting agents have been studied. Earlier,
it was shown that docetaxel, epothilone B
and vinblastine at non-toxic concentrations
efficiently inhibited functional properties of
endothelial cells, while cisplatin and doxorubicin
were not effective. It now appeared that
docetaxel also interferes with actin dynamics
of human ovarian cancer cells, but does not
significantly inhibit microtubule dynamics.
Again, cisplatin and doxorubicin were not
effective. Combining docetaxel with cetuximab
(an inhibitor of EGFR) and trastuzumab (an
inhibitor of HER2) or pertuzumab (an inhibitor
of dimerization of HER2 with EGFR or HER2
with HER3) resulted in increased antitumor
effects in ovarian cancer cells.
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In a project carried out by dr. M. Janmaat,
the focus was on the role of platelet-derived
growth factor (PDGF) signalling in ovarian
cancer. PDGF signalling is not only recognized
as an important process in angiogenesis, as
it is a growth factor for the development of
the supporting cells in the vasculature, the
pericytes, but also as a factor involved in the
tumor-associated fibroblast (stroma) response.
Our data were consistent with such a paracrine
role, while autocrine PDGF signalling in ovarian
cancer cells appeared of no importance for tumor
proliferation and migration.
I n t e g r at i v e
tumor cell biology for
i m p r o v e m e n t o f c a n c e r t r e at m e n t
Starting from the moment of anticancer drug
administration to a patient, many processes
are involved in generating the overall tumor
response to the drug, such as pharmacokinetics,
drug metabolism, drug transport to the target
and interactions of the drug with the cellular
biochemical machinery, including cellular defense
(such as damage repair). Advanced knowledge
on the integration of the dynamic responses of
all these processes will lead to better treatment
and better individual tailoring of anti-cancer
treatments. Parts of these processes have been
mimicked and modelled in vitro using breast
cancer cells in tissue culture and in silico (prof. dr.
J. Lankelma) in a way that simulates treatment of
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breast cancer with doxorubicin.
Another project aims at the design of methods
for monitoring tumor responses dynamically
in patients, because sensitive and selective
biomarkers for this purpose are currently
lacking. To search for new biomarkers in urine,
we are developing a micro-HPLC system
with post-column derivatization, rendering
peptides fluorescent, followed by laser-induced
fluorescence detection. The method has been
applied in five patients with colorectal cancer,
treated with bevacizumab and chemotherapy.
We have collected urine samples two days before
and three weeks after the start of treatment
and we have observed dynamically changing
chromatographic peaks during this time period.
More information on the identity of these peaks
will be obtained by tandem mass spectrometry.
Division
of
G e n e T h e ra p y
Research by the Division of Gene Therapy
focuses on the use of recombinant viruses
as platforms for new anti-cancer therapies.
Currently, the main research aims are to
selectively target cancer cells and to enhance
cancer cell killing by selectively replicating
viruses. The final goal is to translate new
gene-based treatment modalities into clinical
applications in cancer patients. In addition,
the Gene Therapy Laboratory houses the
newly established CCA/V-ICI core facility for
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functional oncogenomics research by RNA
interference library screening (RIFOL).
anti-tumor activity was obtained from bone scan
improvement, measurable tumor regression and
improvement in bone pain. Interestingly, clinical
responses were associated with endocrine-related
immune breakthrough events that were treatable
with standard hormone replacement therapy.
Members of the Division of Gene Therapy.
Gene therapy clinical studies
Currently, one clinical study is ongoing as a
combined effort of the Divisions of Gene Therapy
and Immunotherapy and the Departments of
Urology of the VU University Medical Center and
the Antonie van Leeuwenhoekhuis/Netherlands
Cancer Institute, Amsterdam. This phase I study,
sponsored by Cell Genesys and Medarex, involves
multiple vaccinations with a fixed dose of lethally
irradiated GM-CSF-expressing prostate cancer
cells and dose escalation of a human antibody
against CTLA-4 of patients witt metastatic,
hormone-refractory prostate cancer. This
year, enrolment of the dose escalation cohorts
was completed. Dr. W.R. Gerritsen presented
preliminary results at the 2006 ASCO Annual
Meeting. At higher dose levels, partial PSA
responses were observed. Additional evidence for
x t
In our preclinical studies, we found that the
CRAd Ad5-∆24RGD can be successfully applied
in the treatment of glioblastoma multiforme.
Researchers from the MD Anderson Cancer
Center confirmed the potential of this virus
in an intracranial tumor model. We combined
our efforts with the MD Anderson group and
with prof. D.T. Curiel (University of Alabama,
Birmingham, AL). Jointly, we applied for a
grant from the Rapid Access to Intervention
Development (RAID) program of the U.S.
National Cancer Institute. In 2004, we received
final confirmation of approval of the grant. In
2005, a clinical-grade virus batch was prepared
under the RAID program. This year, toxicology
studies were done and the final report on these
studies is expected in Q1 of 2007. The clinical
study that will be done in Amsterdam will involve
convection-enhanced delivery of Ad5-∆24RGD
into the tumor bed after surgical resection of
recurrent glioma. In the phase I dose escalation
part of the study, we will investigate possible
toxicity of this procedure. Next, we will perform a
modified phase II study to investigate the possible
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anti-tumor efficacy of the treatment. We expect to
start the clinical study in the second half of 2007.
G e n e t h e r a p y p r e c l i n i c a l r e s e a rc h
The main areas of preclinical research of the
Division of Gene Therapy are:
•Tumor-specific targeting of recombinant
viruses
•Augmentation of virus-mediated cancer cell
killing
•Preclinical evaluation of gene-based
(combination) treatments
T umor - specific
targeting of
recombinant viruses
We are exploring the utility of adenoviral vectors
(AdV), conditionally replicative adenoviruses
(CRAds), and recombinant coronaviruses as
anti-cancer agents. To make these viruses most
effective and selective for cancer cells, we target
their infection specifically to surface molecules
that are highly expressed on tumor cells.
Previously, we successfully redirected the binding
of AdV to a variety of tumor cell types by using
adapter molecules that on one side bind to the
AdV capsid and on the other side bind to a cell
surface protein of choice. We also extrapolated
this two-component targeting approach to
CRAds. Recently, we showed that these targeted
CRAds could be made very specific for cells
expressing the target molecule by introducing
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two mutations in virus capsid genes. Currently, we
are using the adapter targeting strategy together
with the Department of Otolaryngology (dr. R.H.
Brakenhoff and H.J.T. van Zeeburg; supported by
the Dutch Cancer Society) to develop targeted
CRAds for early intervention of pre-malignant
oral and oropharyngeal lesions. Potential target
molecules that are highly expressed on squamous
cells were already identified and their utility
to mediate adenovirus infection via antibody
binding was confirmed. In addition, together with
the Virology Division of Utrecht University, we
constructed targeted recombinant murine and
feline coronaviruses for selective lysis of human
cancer cells (dr. H. Verheije and dr. T. Würdinger;
supported by the Dutch Cancer Society). Several
findings provided interesting leads for further
development of recombinant coronaviruses
as anti-cancer agents. Coronaviruses could be
targeted to cell surface molecules by incorporating
genes encoding the appropriate adapter molecules
into their genome. These viruses replicated
in human cells expressing the target receptor
and killed these cells via syncytium formation.
Another interesting observation was that acute
myeloid leukemia cells could be infected via their
Fc receptor using an antibody directed against the
coronavirus spike protein. Most of these studies
are described in the thesis of dr. Würdinger, “The
development of tumor-selective coronaviruses
as anti-cancer agents”, which he successfully
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defended at Utrecht University on February 1,
2006, and for which he received the Greiner
Award of the Dutch Society of Gene Therapy.
Despite the success of targeting via bispecific
adapter molecules, it would be preferred to
develop targeted AdV and CRAds as singlecomponent genetic medicines. This requires
the physical incorporation of specific tumortargeting ligands into the viral capsid. Based on
the close resemblance of the adenovirus fiber
protein and the reovirus attachment protein σ1,
dr. F.H.E. Schagen designed a fiber/σ1 fusion
protein lacking all known native binding sites.
A specific binding ligand (His) for an artificial
receptor molecule (HisR) was incorporated
(Figure 2). Cells expressing the receptor were
transduced up to 50-fold more efficient than
parental cells lacking the receptor. Importantly,
the targeted AdV exhibited 1,000 to 10,000-fold
reduced infection of liver cells and did not bind
to human red blood cells. Moreover, following
systemic administration to mice, the virus
showed prolonged persistence in the circulation
and reduced transduction of all organs tested.
Together with the Division of Immunotherapy
(dr. T.D. de Gruijl and dr. D. Oosterhoff) we are
currently exploiting this targeting platform to
develop AdV targeted towards dendritic cells.
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A ugmentation
of adenovirus - mediated
cancer cell killing
Figure 2. Targeting AdV by genetic modification of
the AdV capsid. (A) Parental 293 cells and derivative
293-HisR cells, expressing an artificial receptor for
His-tagged molecules, were transduced with an AdV
with native tropism (AdGL) or with a similar AdV
carrying His-tagged fiber/σ1 fusion proteins (AdGLHisTar). Transduction was analyzed by fluorescence
microscopy for GFP reporter gene expression. AdGLHisTar efficiently transduced 293-HisR, but not 293
cells. (B) 293-HisR cells were transduced as in A, after
pre-incubating the AdV with monoclonal antibodies
directed against the fiber knob domain or the His-tag
as indicated. Whereas the anti-fiber knob antibody
neutralized transduction by native AdGL, the anti-His
antibody neutralized transduction by targeted AdGLHisTar. This confirmed that targeted transduction was
mediated via the His targeting-ligand.
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CRAds are promising new agents with potential
utility to treat cancer. Adenoviruses kill host cells
by exploiting multiple cell death mechanisms of
which a p53 tumor suppressor protein-dependent
pathway appears to be important for rapid lysis.
As the p53 pathway is dysfunctional in most
cancers this limits the efficacy of CRAd-based
therapies. We therefore set out to develop more
effective oncolytic agents by restoring functional
p53 expression during CRAd replication in
cancer cells (Dr. V.W. van Beusechem; supported
by the Royal Netherlands Academy of Arts and
Sciences). In previous years, we showed that
a CRAd expressing wild type p53 killed most
tested human cancer cell lines, primary tumor
specimens, and tumor xenografts in vivo more
effectively. The anti-cancer effects of CRAdp53 were augmented by chemotherapy and
radiotherapy. We also made two tailored CRAds
for more effective treatment of cancers with
high expression of the p53 inhibitors MDM2 and
HPV E6 protein, respectively. For this, we used
p53 variants that do not bind to these inhibitors.
Recently, we found that CRAd-p53 is also more
effective in combination with the specific
MDM2 antagonist Nutlin-3 (H.C.A. Graat;
Department of Orthopedic Surgery). Together,
these observations showed that oncolytic
adenovirus therapy is most effective if p53 is
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expressed and p53 inhibitors are eliminated.
The biotechnology start-up company ORCA
Therapeutics obtained an exclusive license
on our CRAd-p53 technology. Currently, we
are pursuing the further development of p53expressing CRAds for early clinical evaluation
together with ORCA Therapeutics.
An entirely new way to augment the anti-cancer
potency of CRAds could be by suppressing virus
inhibitory molecules in cancer cells. To this end,
we are exploring the utility of RNA interference
(RNAi). RNAi is a very specific mechanism to
silence gene expression by sequence-specific
mRNA degradation. We already constructed
CRAds expressing short hairpin RNAs that can
trigger RNAi. These CRAds caused efficient
and specific silencing of a target gene in cancer
cells. Currently, we are focusing our attention to
silencing inhibitors of p53. Our final goal is to
identify virus inhibitory molecules in cancer cells
by a functional genomics approach. This year, we
obtained a research grant from the Technology
Foundation STW (dr. V.W. van Beusechem and
dr. W.R. Gerritsen) for this project. This research
will benefit from the initiative to establish the
RNA Interference Functional Oncogenomics
Laboratory (see below).
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P reclinical evaluation of gene - based
( combination ) treatments
To translate new virus-based treatment
modalities into clinical applications in
cancer patients, we investigated the efficacy
of adenoviruses in preclinical models of
osteosarcoma and glioblastoma multiforme.
Dr. M.A. Witlox (Department of Orthopedic
Surgery) completed her studies on the utility of
adenovirus gene transfer vectors and oncolytic
adenoviruses for the treatment of osteosarcoma
(OS) by defending her thesis entitled “Gene
therapy of osteosarcoma. Preclinical studies with
adenoviral vectors and adenoviral oncolysis”on
December 15, 2006. An important finding of
these studies was that the primary receptor for
adenovirus, CAR, is hardly expressed by most
osteosarcomas. Therefore, effective gene therapy
or virotherapy of OS requires targeted virus
entry via alternative cell receptors. In particular,
integrin molecules were found very useful for
this purpose. Because metastatic disease, in
particular to the lungs, is the major threat to
patients with OS, H.C.A. Graat (Department
of Orthopedic Surgery) focused his studies on
treating OS metastases. In collaboration with
prof. E.S. Kleinerman (Division of Pediatrics,
M.D. Anderson Cancer Center, Houston, TX,
USA) he set up a mouse model of human OS lung
metastasis. Intravenous delivery of the integrintargeted CRAd Ad5-∆24RGD to mice with
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established OS lung metastases decreased the
number of lung metastases and total lung weight,
strongly suggesting that systemic treatment of
OS with CRAds is feasible.
Previously, we found that the CRAd Ad5∆24RGD kills glioblastoma cells effectively in
vitro and in vivo. Furthermore, the efficacy of
this treatment was enhanced by combining it
with irradiation. This year, dr. M.L.M. Lamfers
(Department of Neurosurgery) assessed the
effects of these treatments in an intracranial
glioma animal model. Surprisingly, and in
contrast to findings in cell culture and in
subcutaneous glioma tumors, combination
treatment was not significantly more effective
than treatment with virus alone. Apparently,
glioma tumors respond differently to treatments
in their natural brain environment than when
placed in an ectopic environment. Another
interesting observation made by dr. Lamfers
in collaboration with the group of prof.
E.A. Chiocca (Molecular Neuro-oncology
Laboratories, Massachusetts General Hospital,
Charlestown, MA, USA) was that the immune
system appears to reject oncolytic adenovirusinfected glioma cells in the brain. She monitored
virus replication in an intracranial mouse model
of malignant glioma, using a tumor-selective
adenovirus expressing luciferase and in vivo
bioluminescence imaging. Luciferase expression
x t
was lost rapidly, whereas a replication defective
control vector tested in the same model
demonstrated stable luciferase expression. In
immunodeficient NOD/SCID mice, luciferase
expression by the oncolytic adenovirus was
lost less rapidly, suggesting a role for the
immune system. This was corroborated by the
observation of infiltrated inflammatory cells
in injected tumors. Treatment of mice with
the immunomodulator cyclophosphamide,
which decreased tumor infiltration by immune
cells, prolonged transgene expression. Thus,
intracranial anti-tumor efficacy of oncolytic
adenovirus may be hampered by an immune
response against the replicating virus, which
can at least partially be circumvented by
pretreatment with immunomodulating agents.
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RNA I n t e r f e r e n c e F u n c t i o n a l
O n c o g e n o m i c s L a b o r at o r y
On June 19, 2006, during the opening event
of the new research building of the VUmc
Cancer Center Amsterdam (CCA), Dr. V.W.
van Beusechem received a research award from
the Foundation SBGO to introduce a new
platform technology, i.e. RNA interference
library screening. The biological process of
RNA interference (RNAi) provides a wealth
of opportunities for functional oncogenomics
research. Using RNAi, the genetic cause
of a particular function can be assessed by
individually suppressing gene products in cancer
cells and studying changes in the behavior of
these cells. Pinpointing gene products that are
functionally associated with aberrant processes in
cancer cells contributes to a better understanding
of cancer cell biology as well as to identifying
targets for therapy.
To provide researchers at CCA/V-ICI with
facilities to conduct RNAi library screens, a core
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facility was established in the Gene Therapy
laboratories of the Department of Medical
Oncology. This RNA Interference Functional
Oncogenomics Laboratory (RIFOL) provides
researchers access to the Dharmacon siARRAY
Human Genome Library. This library includes
siRNA reagents that target over 21,000 unique
genes of the human genome. RIFOL maintains
equipment to support mammalian cell-based
medium-throughput screening efforts. The
RIFOL user group, with members from several
CCA/V-ICI departments, provides an internal
forum for researchers to share protocols and
exchange screening data. Furthermore, RIFOL
joined the Genome-Wide RNAi Global
Initiative, an alliance of Dharmacon, Inc. and
leading international research centers pioneering
the use of whole-genome RNAi screening.
Interactions with this community of researchers
exploiting this exiting new technology should be
very helpful to advance the productivity of our
research.
Division of
I m m u n o t h e ra p y
The Division of Immunotherapy is a close
collaboration between the Departments of
Medical Oncology and Pathology. Under the
supervision of dr. A.J.M. van den Eertwegh
and prof. dr. R.J. Scheper, the division aims
at translating preclinical studies into new
x t
immunological approaches for the treatment
of cancer. In 2006 the division experienced
considerable growth, both in terms of number of
people and diversity of research subjects.
Members of the Division of Immunotherapy paying
tribute to prof. H.M. Pinedo.
In 2006, dendritic cell (DC)-targeted tumor
vaccination remained a major focus of research,
in which we have a long-standing collaboration
with the division of Gene Therapy (dr. W.R.
Gerritsen and dr. V.W. van Beusechem) and
the Division of Human Gene Therapy of the
University of Alabama at Birmingham (UAB;
dr. D.T. Curiel). Further topics of research
were the suppressive effects of tumors on DC
differentiation and maturation, potentiation of
DC functions in vivo, chemo-immunotherapeutic
approaches, the role of ABC-transporters in DC
physiology, the development of DC lines for
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semi-allogeneic vaccination, and identification
of novel tumor-associated antigens for vaccine
development. The division was also involved
in monitoring of DC and tumor-specific T cell
function in several clinical immunotherapeutic
studies.
Lines of research within the division of
immunotherapy:
• Tumor cell vaccination
• Immunopotentiation
• Dendritic cell-based therapies
• NKT cell-based therapies
Tumor
c e l l va c c i n at i o n
A utologous
tumor cell
vaccination in renal cell carcinoma
In 2004 we started a phase II study in metastatic
renal cell carcinoma, vaccinating patients with
an autologous whole tumor cell vaccine in
combination with Granulocyte-Macrophage
Colony Stimulating Factor (GM-CSF) and PF3512676 (formerly known as CPG 7909). GMCSF recruits and activates myeloid dendritic cells
(MDC), while bacterially derived un-methylated
CpG sequences like PF-3512676 activate
plasmacytoid DC (PDC) and further boost both
natural killer (NK) and cytotoxic T lymphocyte
(CTL) responses. This study was coordinated
by dr. A.J.M. van den Eertwegh. Vaccine
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preparation was performed according to current
Good Manufacturing Practice (cGMP) in our
vaccination facility headed by dr. E. Hooijberg.
The first three induction vaccinations were given
weekly with all subsequent booster vaccinations
given every three months. After completing
tumor cell vaccinations, patients were treated
with subcutaneous injections of PF-3512676
once every two weeks and IFN-α subcutaneous
three times per week with the aim of further
enhancing innate immunity and the effector
phase of the adaptive immune response. Primary
endpoints of this study were response rate and
feasibility, and secondary endpoints were toxicity,
efficacy of immunization and survival. This
year we included the 12th patient in the study,
a patient with metastatic renal cell carcinoma.
All patients tolerated the vaccines very well (no
ulceration) and after 3 vaccinations a significant
DTH response against the autologous tumor
cells was observed in all patients, indicating
that a specific tumor response was induced. In
addition, we observed a partial remission in 25%
of patients. Clinical follow-up is ongoing.
A llogeneic tumor cell vaccination and
anti -CTLA-4 in prostate cancer
A phase I/II clinical trial started last year (led
by dr. W.R. Gerritsen and dr. A.J.M. van den
Eertwegh), studies the anti-tumor activity of a
prostate specific immunotherapy based on two
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Granulocyte/Macrophage-Colony Stimulating
Factor (GM-CSF)-transduced allogeneic cell
lines (Prostate GVAX®) combined with the
immunomodulatory anti-CTLA4 antibody
ipilimumab in men with metastatic, hormonerefractory prostate cancer. The anticipated
synergy of these novel agents may lead to
augmented T cell-mediated anti-tumor immunity
via improved dendritic cell function, and
blockade of inhibitory feedback loops in tumorspecific T cells. Preliminary results showed PSA
responses in 5/6 patients at the two highest
ipilimumab dose levels, as well as regressing
of bone and lymph node metastases, and
improvement of bone pain in three of these five
PSA responders.
S. Lougheed and A. Stam receive a shipment of cell
lines from Cell Genesys Inc. for the GVAX and
ipilimumab immunomonitoring studies.
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Under supervision of dr. T.D. Gruijl and S.J.A.M.
Santegoets, MSc, we are monitoring dendritic
and T-cell function in these patients to identify
changes that correlate with clinical efficacy. The
first results convincingly showed generalized
T cell activation and tumor-specific serological
responses following therapy, in conjunction
with profound decreases in PSA serum levels.
To increase the chances of tumor-specific T
cell detection, both blood and injection sites
are monitored for T cell reactivity. These
analyses are expected to yield valuable data
concerning immune effects achieved by the
GVAX/ipilimumab combination. The first
promising clinical data from the phase I study
were presented by dr. W.R. Gerritsen at the 2006
annual meeting of ASCO in Atlanta. The accrual
of 16 additional patients for the phase II trial is
currently ongoing.
I m m u n o p o t e n t i at i o n
L ocoregional treatment with
GM-CSF and PF 3512676 (CPG7909)
of
melanoma patients
Early melanoma development is accompanied
by impaired immune effector functions in the
initial tumor-draining lymph node, the so-called
sentinel lymph node (SLN). Most notably, a
reduced frequency and activation state of DC
may well interfere with the activation of anti30
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tumor effector T cells. In collaboration with the
Department of Oncological Surgery (prof. dr.
P.A.M. van Leeuwen and dr. S. Meijer), we have
performed two phase II studies of clinically stage
I/II melanoma patients who received either GMCSF or the CpG–B oligodinucleotide (ODN) PF3512676 intradermally around the excision site of
the primary melanoma. Both treatments resulted
in increased frequencies of tumor antigenspecific CD8+ T cells in the corresponding
SLN. The observed rise in melanoma-reactive
CD8+T cells upon GM-CSF administration
was associated with an increased frequency of
(most likely skin-derived) CD1a+CD83+ myeloid
DC in the SLN. In contrast, rises in frequency
of melanoma-reactive CD8+ T cells, observed
upon CpG-B ODN administration in both SLN
and blood, were associated with the activation
of SLN-resident plasmacytoid DC rather than
of CD1a+ myeloid DC. In addition, CpG-B
administration was associated with significantly
lower Treg frequencies as well as the presence of
a newly identified CD11chiCD123+CD83+ mature
myeloid SLN-DC subset, further characterized
by the expression of TRAIL, CCR7, and high
levels of co-stimulatory molecules. These studies
thus provide insight into the different DC
subsets that are present in melanoma-draining
lymph nodes and how these may be targeted in
order to activate tumor-reactive CD8+ T cells.
The latter appear to have been recruited to the
x t
SLN in early stages of melanoma development
but have remained functionally “dormant”.
Clinical strategies aiming at immune modulation
of the SLN may help to (re)activate these tumorreactive T cells and thus afford local as well as
systemic control of metastatic outgrowth.
On September 29, 2006, dr. R.J.C.L.M. Vuylsteke
successfully defended his PhD thesis on the GMCSF studies described above. By the end of 2007,
B.G. Molenkamp is expected to defend a thesis
on the PF-3512676 studies. Meanwhile, B.J.R.
Sluijter, MSc, has just concluded the accrual of
patients in our third phase II trial, comparing
the effects of combined local administration of
PF-3512676 and GM-CSF with the effects of PF3512676 alone.
N eoadjuvant chemotherapy and
GM-CSF in breast cancer
Putative beneficial effects of GM-CSF
administration were also studied in conjunction
with neoadjuvant chemotherapy in patients
with locally advanced breast cancer (LABC)
in an international multicenter phase III trial
(Spinoza Trial, initiated by prof.dr. H.M. Pinedo).
We hypothesized that improved survival might
result from chemotherapy-induced tumor
antigen release in combination with long-term
preservation of the tumor-draining lymph
nodes and GM-CSF-induced DC mobilization
and activation. GM-CSF may correct the
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hampered development and functioning of DC
in these patients while chemotherapy-induced
tumor cell death may decrease the levels of
tumor-derived immunosuppressive cytokines.
These circumstances should facilitate optimal
activation of tumor-specific T cells with the
capacity to eradicate both primary tumor cells
and micrometastases. Phenotypic and functional
DC and T cell studies were performed in
peripheral blood and TDLN biopsies over the
course of treatment by dr. T.D. de Gruijl, S.M.
Lougheed, MSc, and A.M. Stam. We found
significant decreases in myeloid DC frequencies
in the peripheral blood of LABC patients at
the start of treatment, as compared to sex- and
age-matched healthy controls. During treatment
these frequencies increased in LABC patients,
finally even exceeding levels observed in
healthy controls. Myeloid DC frequencies were
significantly increased in TDLN from GM-CSFtreated patients in comparison to G-CSF-treated
patients. Interestingly, CEA, Her-2/neu, and/or
survivin-specific CTL responses were detected
in TDLN in two out of four tested patients, but
not in the blood of the same patients. This is an
indication of local specific CTL priming. The
long-term clinical follow-up analysis is ongoing.
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VEGFR
inhibition :
DC differentiation
Defective systemic DC differentiation,
maturation and function is one of the
mechanisms underlying impaired anti-tumor
immunity in cancer patients. Tumor-derived,
vascular endothelial growth factor (VEGF) has
been implicated as a major DC-suppressive
factor. In a collaborative effort with the Division
of Angiogenesis (dr. K. Hoekman and prof. dr.
E. Boven), H. van Cruijsen, MD, is performing
clinical and preclinical studies to investigate the
potential beneficial effect of VEGF receptor
(VEGFR) tyrosine kinase (TK) inhibitors on
defective DC differentiation in cancer patients.
Peripheral blood DC (PBDC) precursor and
subset frequencies were measured in patients
with advanced renal cell cancer before and after
treatment with the anti-angiogenic, VEGFR-TK
inhibitors AZD2171 or sunitinib. It was found that
a population of immature myeloid cells (ImC) and
a population of myeloid suppressor cells (MSC)
was significantly increased, while more mature
myeloid DC precursors (pMDC) and specifically
the myeloid DC subsets-1 (BDCA-1+/CD1c+) and
-2 (BDCA-3+) were significantly reduced in the
blood of cancer patients as compared to healthy
subjects. Remarkably, plasmacytoid DC (BDCA2+) were also significantly lower. After treatment
with AZD2171, pMDC frequencies did not
increase, while sunitinib treatment did normalize
effects on
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pMDC values in the blood of patients with
advanced renal cell cancer.
ABC transporters and DC functioning
Looking for new ways to optimize DC-based
cancer immunotherapy, we are investigating
possible roles of ATP-binding cassette (ABC)
transporters in physiological DC processes
such as differentiation and migration. These
ABC transporters are involved in multidrug
resistance (MDR) in patients undergoing
chemotherapy. Knowledge of the possible role of
these transporters in immune effector functions
may aid in the rational design of future chemoimmunotherapeutic approaches. R. van de Ven,
MSc, works on a NKB-funded project. From
April to August 2006, she visited the laboratory
of prof. dr. D.T. Curiel to construct adenoviral
vectors encoding siRNAs for various ABC
transporters. These studies have so far uncovered
roles for MRP1 in DC differentiation, for MRP4
in DC migration and for BCRP in Langerhans
cell (LC) differentiation. Modulation of the
expression of these ABC transporters to enhance
DC development and function (e.g. by cytostatic
drugs) may be a novel way to support DC-based
tumor immunotherapy approaches. R. van de
Ven received a Young Investigator’s Award for
this work at the 2006 Annual Meeting of the
American Association for Cancer Research
(AACR), held in Washington DC.
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cell-based therapies
situ targeting and modulation of
dendritic cells for immunotherapy :
reversal of suppression
Vaccination with DC generated in vitro holds
great promise, but is hampered by suboptimal
migration of DC to the draining lymph nodes.
Direct in vivo targeting of vectors carrying the
genetic code of tumor antigens to DC may thus
present an attractive alternative. Unfortunately,
in cancer patients, hampered DC differentiation
and activation has been reported. High systemic
levels of tumor-derived suppressive cytokines
like IL-10, IL-6 and VEGF have been shown
to interfere with proper DC function, resulting
in tolerance rather than immune activation.
In a Zon/MW-funded VIDI project awarded
to dr. T.D. de Gruijl, dr. D. Oosterhoff and J.J.
Lindenberg, MSc, are investigating ways to target
DC in situ, while bypassing or reversing tumorinduced immune suppression.
A human skin explant model is used to study
effects of suppressive factors on DC migration
and antigen presentation. A shift from mature
CD83+ DC to immature CD14+BDCA3+
macrophage-like cells was observed within
seven days after migration from IL-10 injected
skin. These macrophage-like cells displayed
poor T cell-stimulatory ability and lacked
32
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expression of CCR7, thus precluding their
migration to T cell areas in lymph nodes. Of
note, DC migrating from breast tumor-overlying
skin samples underwent a similar accelerated
DC-to-macrophage switch. Recent studies
have demonstrated that hyperactivation of
the Jak2/STAT3 pathway by the suppressive
cytokines IL-10 or IL-6 might be responsible for
hampered DC differentiation in cancer. In order
to prevent (or revert) abnormal differentiation
into suppressive DC in cancer patients, we have
constructed a panel of replication deficient
adenoviral (Ad) vectors expressing both GFP and
short hairpin RNAs directed against different
key proteins of the Jak/STAT pathway, such
as STAT3. These adenoviral vectors are used
to transduce suppressed DCs; effects on DC
phenotype and function are being studied.
In order to identify useful targeting motifs
on DC in human skin explants and tumordraining lymph node samples, we will use a
screening system based on Ad vectors in which
a Staphylococcus aureus protein A domain is
incorporated, that can bind the Fc domain
of immunoglobulins. This system allows the
utilisation of antibodies directed against DC
surface markers to test their efficacy as targeting
molecules for high-efficiency transduction and
simultaneous activation of the targeted DC. In
collaboration with Crucell (dr. M. Havenga), we
x t
found Ad35 to efficiently transduce maturing
DC in the dermal tissue environment through
binding to CD46. In a collaborative study with
UAB (dr. D.T. Curiel), R. van de Ven and J.J.
Lindenberg, MSc, are similarly investigating DC
targeting potential of Ad5/3 vectors that bind
CD80 and CD86.
In a related NKB-funded project, dr. B.
Hangalapura and P.G.J.T.B. Wijnands, MSc, are
investigating the utility of CD40-targeted Ad
vectors for in vivo DC-based tumor vaccination.
The interaction of CD40, a cell surface marker
expressed by DC, and its ligand is critical for the
generation of protective cell-mediated tumor
immunity. We used CFm40L, a bispecific adaptor
molecule with the ectodomain of CAR on one
end linked to the extracellular domain of mouse
CD40 ligand on the other end by a trimerization
motif (provided by dr. A. Pereboev, UAB). This
adaptor molecule bridges the fiber knob domain
of Ad to CD40 on both mouse (m) and human
(hu) DC. Re-targeting of Ad to CD40 using
CFm40L enhances adenoviral infection and also
induces phenotypic and functional maturation
of DC. Melanoma antigen recognized by T
cell-1 (MART1) and tyrosinase related protein2 (TRP2) are recognized by melanoma specific
CTLs. Using CFm40L-targeted Ad encoding
huMART1 to infect human DC in vitro, superior
priming ability of fully functional CTL was
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established. We are currently using CFm40Ltargeted Ad encoding mTRP2 to target DC in
vivo and examine the effects in terms of T cell
activation and tumor protection in the B16
melanoma model.
D evelopment
of the human
MUTZ-3
dendritic cell line model for
semi - allogeneic tumor vaccination
In a separate line of research the human acute
myeloid leukemia (AML) cell line MUTZ-3
and newly generated AML lines are developed
to obtain DC in vitro. These DC will be
used as allogeneic, standardized off-the-shelf
alternatives to autologous DC for DC-based
tumor vaccination. Together with the VUmc
spin-off DCPrime (prof. dr. A.M. Kruisbeek
and prof. dr. R.J. Scheper), we are currently
planning phase I clinical trials, in which patients
with AML, glioblastoma or prostate cancer
will be vaccinated with MUTZ-3 derived DC
(either unloaded, fused to glioma cell lines
or pulsed with prostate tumor RNA), as an
HLA-A2/A3-matched allogeneic vaccine. In
preclinical proof of principle studies, we have
recently shown that DC can be generated from
the CD34+ MUTZ-3 cells. CD14+ MUTZ-3
precursors can either differentiate to Langerhans
cells (LC; displaying typical Birbeck granules;
Figure 3), or to interstitial DC. To assess the
utility of these MUTZ-3-derived DC subsets
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for immunotherapeutic applications, we tested
their potential to migrate to paracortical lymph
node areas, their ability to prime tumor-specific
CTL, and their cytokine release profile. From
these studies, we concluded that the MUTZ-3
cell line may provide a continuous supply of DC
and LC precursors for tumor immunotherapy
approaches. Before the end of 2007, S.J.A.M.
Santegoets is expected to finish her PhD thesis
based on these studies.
Figure 3. Birbeck granules provide evidence that
MUTZ-3 cells can differentiate into bona fide
Langerhans cells.
NKT cell-based therapies
Key features of invariant natural killer (iNKT)
cells are an invariant T cell receptor gene
usage, CD1d restriction, high levels of cytokine
production, particularly interleukin-4 (IL-4) and
interferon-γ (IFN-γ), and cytotoxic potential.
x t
Animal studies showed that iNKT cells,
depending on their predominant cytokine profile,
are important regulatory cells in autoimmunity
(Th2, IL-4) and tumor immunity (Th1, IFN-γ).
iNKT cells recognize hydrophobic ligands like
α-galactosyl-ceramide (α-GalCer), in conjunction
with the antigen presenting molecule CD1d.
Under supervision of dr. B.M.E. von Blomberg,
head of the Laboratory for Medical Immunology,
the size of the circulating iNKT cell population
in various types of cancer was compared with
matched healthy volunteers. This population
was found to be decreased in cancer patients.
Whereas the number of T cells or NK cells had
no prognostic value, patients with a strongly
diminished iNKT cell fraction prior to therapy
had significantly decreased disease-specific
survival. We are currently developing strategies to
increase the frequency of iNKT1 cells in cancer
patients, which will hopefully enhance their antitumor immunity. One such approach involves
treatment with the ligand α-GalCer, systemically
or locally by intradermal administration. Adoptive
transfer of iNKT cells expanded in vitro is an
alternative approach, the validity of which was
tested in a murine tumor model. In a NKBfunded study, dr. H.J. Bontkes and J.W. Molling,
MSc, developed a culture method for the
expansion of spleen-derived murine polyclonal
iNKT cell lines. These iNKT cells could elicit
anti-tumor responses in vivo; tumor outgrowth
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was inhibited after intradermal co-injection
with B16.F10 melanoma tumor cells and the
formation of lung metastases was inhibited after
intravenous co-injection with B16.F10. These
data show that cultured murine iNKT cells retain
their function and that adoptive transfer of iNKT
cells may indeed enhance anti-tumor responses.
J.W. Molling is currently finishing his PhD thesis
based on these studies. Dr. H.J. Bontkes and M.
Moreno, MSc, are currently investigating the
influence of iNKT1 cells on tumor-reactive NK
and CTL activation and expansion to define the
mechanism by which metastasis formation was
inhibited.
F u r t h e r c o l l a b o r at i v e p r o j e c t s
•Development of tissue engineered, autologous
human skin for studying sensitization
and immuno-vaccination approaches: in
collaboration with the Department of
Dermatology (dr. S. Gibbs) human, fully
defined, immuno-competent in vitro skin
models (containing Langerhans cells) are
developed by K. Ouwehand, MSc, and
employed to study immunogenicity of vaccine
formulations and allergens.
•Optimisation of strategies for DC vaccination
in AML: in collaboration with the Department
of Hematology (dr. A. van de Loosdrecht and
prof. dr. G. Ossenkoppele) novel approaches to
DC-based AML vaccination are explored.
34
•CTL activation markers as prognostic factors
in melanoma: in a collaboration with the
Department of Pathology (dr. J.J. Oudejans)
intratumoral or intranodal CTL activation
and apoptosis (resistance)-related markers
are studied by I. van Houdt, MSc, for their
possible significance in melanoma progression.
Division
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of
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her PhD studies in the group of prof. Peters
funded by an exchange grant.
P h ar m ac o l o g y
In the Division of Pharmacology research is
mainly focused on the mechanism of action
of, and resistance to, cytotoxic and novel
targeted anticancer agents in relation to
pharmacogenomic, pharmacodynamic and
pharmacokinetic parameters. This research
was coordinated in the laboratory by prof.
dr. G.J. Peters and in the clinic by prof.dr. G.
Giaccone and dr. C. van Groeningen. Dr. F.A.E.
Kruyt supervised the research on cell death
mechanisms, dr. J. Rodriguez on targeted agents
in the laboratory and prof.dr. Giaccone in the
clinic. Following current clinical practice in
medical oncology, research on several targeted
agents is becoming more and more integrated
in all research lines. In this year, the division
hosted a number of foreign scientists working in
the various research groups, in particular Elisa
Giovannetti, PhD, from Italy, Cecilia Ceresa,
MSc, from Italy, Yerigeri Mayur, PhD, from
India, and Christina Tekle, MSc, from Norway,
while Clara Lemos, MSc, from Portugal performs
Members of the group of prof. Peters
Clinical
p h a r m ac o k i n e t i c s a n d
p h a r m ac o d y na m i c s
The flavonoid monoHER has been investigated
for its protective effects against doxorubicininduced cardiotoxicity. Scheduling of monoHER
was investigated in an animal model by A.
Bruynzeel, MD, while in a clinical study the
potential protective effects of monoHer in
humans were investigated by taking biopsies
after repeated administration of doxorubicin
with monoHER. Differences in metabolism
of the flavonoid between humans and rats
due to conjugation and sulfatation have been
investigated.
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Application of liquid chromatography (LC)
coupled to mass-spectrometry (LC-MS-MS)
continued to increase. A sensitive LC-MS-MS
detection method for gemcitabine, developed
by dr. R. Honeywell, was used to measure the
low plasma concentration of gemcitabine (50150 nM) after administration as a hepatic artery
infusion, and (phosphorylated) gemcitabine in
brain tumors. These data demonstrated for the
first time that gemcitabine passes the bloodbrain/tumor barrier. The same method was
applied in the quantification of deoxycytidine
(CdR) in various clinical schedules. These
studies demonstrated that gemcitabine
treatment resulted in an increase of CdR.
Other applications of LC-MS-MS include the
measurement of folates in patients enrolled
in a randomized phase II study, comparing
gemcitabine-cisplatin with or without folate
supplementation. This study was initiated in
2005 and patients are still being entered. The
LC-MS-MS method also enabled measurement
of the accumulation of deoxyuridine after
administration of the thymidylate synthase (TS)
inhibitor pemetrexed (Alimta®). These data are
being related to the clinical effects of treatment.
A small adaptation of this assay allowed
the analysis of DNA methylation patterns.
Methylation of deoxycytidine in the promoter
region of individual genes will effectively turn off
gene function. The degree of methylation was
35
investigated in several cell lines and in paired
cells of colon tumors and normal mucosa.
T r a n s l at i o n a l
Ne
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r e s e a rc h ,
p h a r m ac o g e n o m i c s
Pharmacogenomic features of various
populations are considered increasingly
important as a tool for selecting the most
optimal treatment for patients, aiming at
reducing toxicity and/or increasing antitumor
activity of an anticancer agent. The study in
which optimal treatment was based on the
tumoral expression of thymidylate synthase (TS)
and dihydropyrimidine dehydrogenase (DPD),
the TS/DPD study, is now being followed by a
retrospective study of historic samples using
micro-array analysis of the whole genome, and
array-CGH analysis of the DNA, in collaboration
with dr. B. Ylstra and prof. G.A. Meijer of the
Department of Pathology. In a small cohort of
15 patients, non-supervised clustering of the
array-CGH data revealed a significant difference
between responding and non-responding
patients.
Further pharmacogenomics research focuses on
enzymes that play an important role in folate
homeostasis. Since it is known that a 677-TT
homozygosity of methylene tetrahydrofolate
reductase (MTHFR) is associated with lower
enzyme activity, this polymorphism will increase
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the concentration of the substrate of TS and may
affect the efficacy of drugs targeting TS, such
as 5-fluorouracil and pemetrexed. Therefore,
polymorphism of MTHFR together with
that of other folate-related enzymes has been
determined in normal and tumor tissues of
patients being treated with antifolates. In colon
cancer patients, an association between MTHFR
polymorphism and response was observed.
A ntimetabolites , DNA
repair ,
cell cycle antagonists and
multidrug resistance proteins
Antimetabolites interfere with cellular
functioning at many levels. Not only do
they compete with normal metabolites for
incorporation into RNA or DNA, affecting
cellular proliferation, they also directly affect
other processes such as cell cycle regulation
and angiogenesis. Thus, the antimetabolites,
methotrexate and 5-fluorouracil (5-FU) appear
to have additional mechanisms of action, which
involve interference with signalling. Vice versa,
alterations in cellular signalling processes can
profoundly affect the antitumor activity of these
agents. For instance, the activity and expression
of thymidine phosphorylase (TP), also known
as platelet-derived endothelial growth factor
(PD-ECGF), is controlled by cytokines such as
IL-8. TP plays a major role in cellular thymidine
homeostasis; one of the products of this
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reaction, deoxyribose-1-phosphate (dRib-1-P), is
thought to play a major role in angiogenesis. TP
also catalyses the last and rate-limiting step of
the activation of capecitabine (Xeloda®) to 5-FU,
namely that of 5’deoxy-5-fluorouridine (5’DFUR)
to 5-FU. Inhibition of TP may prevent tumorspecific angiogenesis. Irene Bijnsdorp, MSc,
therefore investigated the downstream effects
of TP inhibition both in tumor cells and human
umbilical vein edothelial cells (HUVECs). The
first data indicate differential effects in these
cell types, opening up possibilities for selective
treatment with combinations of TP inhibitor
(TPI) such as the formulation TAS-102, which
consists of trifluorothymidine (TFT) and TPI.
Overexpression of drug transporters may confer
resistance to a number of unrelated drugs.
Previously it was demonstrated that folates are
substrates of MRPs, but also that the expression
of MRPs is affected by folate supplementation
or depletion. Clara Lemos, MSc, demonstrated
that folate homeostasis modulates breast cancer
resistance protein (BCRP or ABCG1) and
multidrug resistance protein 1 (MRP1) expression
differently in cell lines from various origins. The
change in BCRP expression was accompanied
by loss of resistance to mitoxantrone in MCF7/MR LF cells and induction of mitoxantrone
resistance in Caco-2 LF cells. Interestingly, it
was found that BCRP localisation was either
36
cytoplasmic (Caco-2 cells) or in the plasma
membrane (MCF-7/MR cells). Only in the
latter cells, folate deprivation induced loss of
BCRP expression. A visiting scientist from
India, Y. Mayur, PhD, investigated whether N10substituted fluoro-acridone derivatives might
be a substrate for one of the efflux pumps Pglycoprotein, MRP1-5 or BCRP.
I nteraction
Ne
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of cytotoxic
drugs with targeted agents
In order to increase selectivity of cancer
treatment, convential cytotoxic treatment is
increasingly being combined with newer drugs
interfering with cellular signal transduction
pathways. Cellular proliferation and functioning
is controlled by growth factors which activate
membrane proteins by phosphorylation, followed
by a cascade of phosphorylation reactions inside
the cell.
Erlotinib (Tarceva®), a potent small molecule
inhibitor of the tyrosine kinase domain of
epidermal growth factor receptor (EGFR)
was combined with both the platinum analog
satraplatin (C. Ceresa , MSc) and the antifolate
pemetrexed (E. Giovannetti, MD). Pemetrexed
was also combined with the protein kinase C-β
(PKC-β) inhibitor enzastaurin (C. Tekle, PhD).
The active metabolite of satraplatin, JM118,
and erlotinib appeared to be synergistic in
non- small cell lung cancer (NSCLC), colon
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and ovarian cancer cell lines by interfering with
p-ERK and p-AKT signalling downstream of
EGFR, while p38 signalling was not modulated.
The interaction between erlotinib and
pemetrexed was investigated in six NSCLC
cell lines, characterized by a large variation
in thymidylate synthase (TS) expression, the
target of pemetrexed, and by the presence of
mutations of EGFR, K-ras, and PTEN, all
determinants of erlotinib sensitivity. Synergistic
cytotoxicity was detected in all cell lines,
mostly with the pemetrexed (24h) → erlotinib
(72h) sequence, associated with a significant
induction of apoptosis. Pemetrexed increased
EGFR phosphorylation and reduced AKT
phosphorylation, while erlotinib significantly
reduced TS activity, which was additionally
reduced by the combination. In two of the
cell lines, the interaction of pemetrexed with
enzastaurin was investigated, showing enhanced
pemetrexed- induced cell death by enzaustarin.
Enzastaurin reduced the phosphorylation of
targets in cell signalling pathways such as AKT
and GSK3b as well as TS expression, thus
potentially enhancing pemetrexed activity.
T argeting
apoptosis and
kinase pathways for cancer therapy
Research by the group of dr. F.A.E. Kruyt and
prof. G. Giaccone ranges from fundamental
to clinical research on the mechanisms that
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determine life-and-death decisions in tumor
cells. The decisive step in the action of standard
anticancer drugs and novel targeted agents
is their ability to induce molecular pathways
in tumor cells that lead to cell death by
activation of apoptosis. For this reason, the
gain in fundamental knowledge of molecular
mechanisms underlying cell death regulation is
key for the rational design of novel treatment
strategies and the development of targeted
medicines. In addition, molecular pathways
that regulate apoptosis are often defective in
tumor cells and are likely to contribute to drug
resistance and poor patient survival. Hence,
apoptotic regulators may have diagnostic and
prognostic value. Parallel to the preclinical work,
clinical studies with novel targeted agents are
conducted.
Members of the group of prof. Giaccone and dr. Kruyt
37
C onventional
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chemotherapy
and apoptosis
We have discovered that in chemotherapyresistant non-small cell lung cancer (NSCLC)
cells, the apoptotic pathway headed by caspase-9
is blocked. Caspases are a group of proteases that
execute the apoptotic program. A. Checinska,
MSc, supported by a grant of the Dutch Cancer
Society (NKB), is exploring the mechanism
underlying caspase-9 inhibition. She will defend
her thesis in March 2007. In this context, the
involvement of members of the inhibitor-ofapoptosis protein family (IAPs) that are able
to bind to, and inhibit caspases, is evaluated.
In her studies, a potential small molecule drug
that is able to mimic the activity of Smac (a
kind gift from dr. X. Wang, Dallas) is used.
Smac is a cellular protein that can inactivate
IAPs. Also mass spectrometry-based proteomic
strategies are employed to identify caspase-9
binding proteins in NSCLC cells, studies that are
conducted in collaboration with dr. C. Jiménez,
VU University, Amsterdam).
L. Bröker, MSc, has finished her studies on the
mechanisms underlying apoptosis activation by
microtubule-interacting agents. She will defend
her thesis in 2007. One of the main findings in
her studies was that these potent anticancer
drugs induce apoptosis via caspase-independent
pathways. In particular, the lysosomal protease,
cathepsin B, was found to mediate cell killing by
x t
the microtubule-stabilizing agents, paclitaxel and
epothilone B.
T argeted medicines
Based on our mechanistic studies, we
hypothesize that agents that can restore caspase9 activation or that bypass the blockade will
have therapeutic benefit in NSCLC. Results by
J. Voortman, MD, MSc, supported by a NWOAGIKO grant and A. Checinska showed that
the proteasome inhibitor, bortezomib, (PS-341,
Velcade®) is able to restore caspase-9 activation
in NSCLC cells. Apart from the preclinical work,
J. Voortman also coordinated a clinical study in
which the efficacy of this agent, in combination
with other anticancer agents, in the treatment
of solid tumors is examined. As a means to
bypassing caspase-9, current research is focusing
on tumor necrosis factor (TNF)-related apoptosis
inducing ligand (TRAIL) that activates caspase8-dependent apoptosis. Two PhD students
supported by a Dutch Cancer Society grant
(in collaboration with dr. S. de Jong and prof.
dr. E.G.E. de Vries, University Medical Centre
Groningen) are currently working on the efficacy
and mechanism of action of combinations of
TRAIL and bortezomib in NSCLC models (A.
Watts, MSc, in Amsterdam and J. Stegehuis in
Groningen).
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Dr. F.A.E. Kruyt obtained a grant from the
Top Institute Pharma (TIPharma), a national
governmental initiative to stimulate the
interaction between industry and academic
centres. TIPharma aims at achieving leadership
in research and education in areas critical
for the international competitiveness of the
pharmaceutical industry in the Netherlands.
Together with the academic groups of dr. S. de
Jong and prof. dr. E.G.E. de Vries, and prof. dr.
W. Quax (University Medical Centre Groningen),
prof. dr. J. Borst (Netherlands Cancer Institute,
Amsterdam), prof. dr. J.P. Medema (Academic
Medical Center, Amsterdam), a collaboration
has been established with Organon (Oss) and
Pepscan (Lelystad) to study TNF ligands in
cancer. At the VU University Medical Center, we
will in particular study newly developed agonistic
TRAIL receptor-directed monoclonal antibodies
for anticancer activity, and determine the
involvement of kinase pathways in modulating
the response.
Signaling induced by the epidermal growth factor
receptor (EGFR) is known to affect survival
of NSCLC cells. Agents targeting EGFR have
provided novel promising therapeutic strategies.
M. Janmaat, MSc, studied the mechanism and
efficacy of EGFR inhibitors, including the
monoclonal antibody C225 (cetuximab) and the
small molecule EGFR-tyrosine kinase inhibitors
38
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gefitinib (Iressa®) and erlotinib (Tarceva®). He
found that their efficacy was related to the
activation status of the MEK/Erk (MAPK) and
PI3K/Akt kinase pathways. In October 2006, M.
Janmaat successfully defended his thesis entitled
“Targeting Erb receptors as anticancer therapy:
factors of sensitivity and resistance”.
The recent discovery that mutations in the
ATP binding site of EGFR confer sensitivity to
small-molecule EGFR inhibitors in advanced
NSCLC patients has led us to the sequencing of
tumor samples of patients enrolled in phase II
studies of EGFR inhibitors in esophageal cancer
and NSCLC. A grant from the Dutch Cancer
Society (NKB) has been obtained for this work.
M. Gallegos-Ruiz, MSc, is spearheading this
work. In addition to setting up the techniques
to detect mutations in EGFR and K-Ras, we are
performing an extensive analysis of a large series
of NSCLC samples. Using tissue microarrays,
the expression of several proteins in the EGFR
pathway and related signaling networks are being
examined. Furthermore, expression microarrays
and array comparative genomic hybridization
(aCGH), in collaboration with dr. B. Ylstra, are
being carried out to gain a better understanding
of the genetic changes that occur during NSCLC
development.
Dr. Maarten Janmaat successfully defended his thesis; Laudatio by prof. Giaccone
R egulation
of inhibitors of
(IAP s )
Research by dr. J. Rodriguez, supported by the
Walter Bruckerhoff Stiftung, focuses on the
functional implications of regulated nuclearcytoplasm shuttling of proteins that regulate
key cellular processes, such as apoptosis and
cell cycle progression. Current research focuses
on survivin, a member of the IAP family with
a dual role in apoptosis and mitosis. Survivin
shuttles between the nucleus and the cytoplasm.
Several splice variants of survivin, which undergo
differential transport and localize to different
cellular compartments have been identified. The
role of each variant in tumor development and
apoptosis proteins
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progression is being investigated. Using RNA
interference and mutagenesis, we are exploring
several aspects of survivin biology and its relation
with other proteins, such as the kinase aurora
B, both during mitosis and interphase. On the
other hand, using novel imaging techniques, such
as bimolecular fluorescence complementation
(BiFC), the dimerization ability of survivin in
vivo is being explored as a potential target for
anticancer therapies. In this regard, we have
recently identified a functional role for survivin
dimerization: regulation of its nuclear export.
These studies are carried out in collaboration
with dr. R. Medema and dr. S. Lens (University
Medical Center Utrecht) and with the group of
dr. M. Fornerod (Netherlands Cancer Institute,
Amsterdam).
B. Vischioni, MSc, obtained her PhD degree
on November 14, 2006 after defending her
thesis entitled “Expression and subcellular
localization of inhibitor of apoptosis (IAP)
proteins in human tissues: mechanisms and
clinical implications”. She studied the regulation
of the nucleo-cytoplasmic localization of the
survivin relatives cIAP1, cIAP2 and XIAP.
She has found that cIAP1 and cIAP2, but not
XIAP are nucleocytoplasmic shuttling proteins.
She mapped several CRM1-dependent nuclear
export signals (NES) in these IAPs. Extending
these mechanistic studies, B. Vischioni has also
39
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investigated the distribution of IAPs in normal
tissues and tumor samples. She has also explored
the expression of the kinase aurora B, a partner
of the IAP survivin in NSCLC tumor samples
and the effect of novel aurora B inhibitors on the
growth of NSCLC cells in vitro. These studies
are essential to predict side effects in patients
undergoing treatment with IAP inhibitors, drugs
that are presently entering clinical trials.
P r o g n o s t i c m a r k e r s i n NSCLC
The prognostic value of expression of the
apoptotic regulator TUCAN has been studied
by A. Checinska. Although expression did
not correlate with treatment outcome in
NSCLC patients, cytoplasmic localization of
TUCAN predicted shorter survival. Dr. F.
Barlesi, a guest from the Assistance Publique
Hopiteaux de Marseille, France, studied global
histone modifications as prognostic factor for
resected NSCLC patients. His data indicate
that methylation and acetylation of histone H3
are valuable prognostic markers for a subset of
early-stage NSCLC patients. Another visitor
in our department, dr. P Zucali from Italy,
studied the relationship between activation
of the c-Met receptor pathway and response
to EGFR inhibitors in NSCLC. Using tissue
microarrays and in vitro experiments, he
found that activation of c-Met may be marker
of intrinsic resistance to gefitinib and that
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combined inhibition of the c-Met and EGFR
pathway may be a suitable therapeutic approach
for some NSCLC patients. These studies were
performed in collaboration with prof. G. Meijer,
prof. P. van der Valk and dr. J. Oudejans from the
Department of Pathology.
O n c o P r o t e o m i cs
L a b o rat o r y
CCA
O
ONCO
VUmc
PL
Proteomics Laboratory
The OncoProteomics Laboratory (OPL) has
been founded in April 2006 together with the
establishment of the cancer research building of
the VUmc Cancer Center Amsterdam (CCA).
OPL was created to provide a state-of-the-art
proteomics infrastructure and knowledge center
for CCA/V-ICI researchers.
Proteomics is a relatively new field. It creates a
link between genomic information and biological
function through global studies of protein
expression, protein modification and proteinprotein interactions. Until now, proteomics has
been a technology-driven science. Emphasis in
the coming years will be on applying proteomics
to the understanding of biological function in
V U m c M e d i c a l O n c o l o g y – A n n u a l R e p or t 2 0 0 6
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healthy organisms and in disease (Figure 4).
to high-throughput MALDI-TOF/TOF mass
spectrometry, and 2) label-free (nano)-LC-LTQFT mass spectrometry of sub-fractionated
biofluids and tumor samples for in-depth
profiling.
Code
Readout
DN A
~22.000
R NA
~100.000
R e s e a rc h
x t
s t r at e g y
A. D evelopment
of robust automated
mass spectrometry - based methods for
biomarker discovery , with a focus
on subproteomes
P R OT E IN
> 500.000
W ork ing m a c hine ry
Figure 4. Approximation of the number of genes,
messenger RNA molecules and proteins in humans,
illustrating the complexity of the proteome
M i s s i o n o f OPL
The mission of OPL is to develop and implement
innovative proteomics technologies and data
analysis methods to improve diagnosis and
treatment of cancer. To this end, the major
focus is on developing and implementing robust
strategies for biomarker discovery in tumor
tissue, if available, and in biofluids such as
blood-serum/plasma that can be collected noninvasively as well as on proximal fluid such as
CSF and nipple aspirate fluid. In addition, cancer
cell conditioned media and tumor secretomes
hold great promise for discovery of candidate
biomarkers. Samples are profiled using two
complementary platforms for the discovery
of diagnostic, predictive and drug response
patterns and biomarkers: 1) an automated
magnetic bead peptide capture method coupled
40
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O r g a n i s at i o n o f OPL
OPL is a facilitating center where most
projects are shaped in close interaction with
the collaborators. Moreover, a substantial part
of the activities is core research to develop
and implement proteomics methods of general
interest for cancer researchers and clinicians.
Head of the unit is a scientist/coordinator
who is leading a team of scientists (a mass
spectrometrist, a computer scientist and
a biochemical technician as well as some
technicians on collaborative temporary projects).
OPL is housed by the Department of
Medical Oncology. The head of OPL is
assisted by a ‘Program Advisory Committee’
with representatives from the major V-ICI
departments (Medical Oncology, Hematology,
Pathology, ENT, Molecular, Cellular Biology and
Immunology, and Epidemiology and Biostatistics)
who meet every two months with the head of
OPL to discuss and assist in prioritizing projects.
The lack of reliable, robust and easily assessable
biomarkers greatly hampers cancer management.
In recent years, proteomics has raised the hope
of identifying novel biomarkers for cancer
diagnosis and detection. This hope is based
on the ability of proteomic technologies, such
as mass spectrometry, to identify hundreds of
proteins in complex biofluids.
OPL focuses on two complementary approaches
for the discovery of diagnostic and predictive
peptide patterns and identification of candidate
biomarkers in body fluids and tumors (Figure 5).
The first goal was to implement robust, scalable
methods for patient sample preparation that can
be performed in parallel and in an automated
manner.
Pre-analytical variables of serum sample handling
and analytical aspects of the high through-put
(HTP) profiling workflow have been investigated
(Jiménez et al., work submitted and in
preparation). Therefore, the automated method
can now be applied to cancer profiling studies.
In several phase I studies of the Department of
Medical Oncology, serum for proteomics analysis
has been collected.
V U m c M e d i c a l O n c o l o g y – A n n u a l R e p or t 2 0 0 6
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OPL focuses on two complementary approaches for the discovery of diagnostic and predictive peptide
patterns and identification of candidate biomarkers in body fluids and tumors (Figure 5). The first goal
was to implement robust, scalable methods for patient sample preparation that can be performed in
P parallel
r e v and
i ou
s automated manner.
Contents
in an
HIGH-THROUGHPUT
POTENTIAL
DIAGNOSTIC TOOL
IN-DEPTH
HIGH RESOLUTION
& DYNAMIC RANGE,
IDENTIFIED
BIOMARKERS
Patients
M AS S
SP E CT RO M ET R Y
Peptide
mass
profile
Multi-dimensional
Proteomic
profile
HTP
In-depth
Pattern recognition
M AL D I-T OF- M S
100
Diseas
e
% intensity
165
100
5
182
6
199
7
216
8
165
5
182
6
199
7
216
Disease protein
Signature
Retention
time (min)
233
9
control
Mass (m/z)
8
233
9
Nano- LC -L TQF TM S
Novel biomarkers
DIAGNOSIS
PROGNOSIS
DRUG RESPONSE
Diseas
e
Contro
l
Mass (m/z)
Figure 5. Complementary mass spectrometry-based proteomics approaches for discovery of cancer signatures and
Figure
5. Complementary
mass spectrometry-based
approaches formethod
discovery
of cancer
signatures
biomarker
at OPL.
LEFT: High-throughput
MALDI-TOFproteomics
mass spectrometry-based
coupled
to spectral
data
and biomarker at OPL. LEFT: High-throughput MALDI-TOF mass spectrometry-based method coupled to
mining of blood-serum peptides and proteins for the discovery of predictive, diagnostic and drug-response signatures.
spectral data mining of blood-serum peptides and proteins for the discovery of predictive, diagnostic and drugTissue response
homogenates
can also Tissue
be subjected
to this type
analysis.
RIGHT:
sub-fractionation
method
coupled
signatures.
homogenates
canofalso
be subjected
toProteome
this type of
analysis. RIGHT:
Proteome
subto (nano)Liquid-Chromatography-LTQ-FT mass spectrometry for in-depth profiling of peptides and proteins and
discovery/identification of predictive and early detection biomarkers in patient samples. Subproteomes of interest are
37 the nucleus and secreted proteins. In blood, the low
various subcellular tissue fractions including the plasma membrane,
molecular weight proteome as well as the glycoproteome are of particular interest.
41
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Robust methods for the capture of subproteomes
from tissue and blood for in-depth profiling
by nanoLCFTMS and discovery of identified
candidate biomarkers are currently being
developed and explored in several (collaborative)
projects (among others with dr. F. Kruyt , dr.
H.J. Broxterman, dr. K. Hoekman and with
dr. R.Fijneman, prof. dr. G. Meijer and dr. B.
Carvalho of the Department of Pathology).
B. D ata analysis
We are developing robust data analysis methods
for label-free quantitation of peptide ion
abundance, because this approach represents
a promising (and economic) avenue for
quantitation at high sensitivity. The additional
dimension of separation allows for profiling of
a much larger number of peptides in complex
mixtures. We are implementing open-source
tools as well as home-made algorithms (Horizon
Breakthrough project granted to Drs. Marchiori
and Jiménez) for the analysis of LC-MS datasets.
The sensitivity of LC-MS profiling is higher, but
the throughput of this method is considerably
less than the MALDI-TOF-MS approach (~ 10
samples per day), so we will use high quality,
well-characterized samples for this approach.
Statistically significant differential peptides
will be followed up in targeted tandem MS
experiments to determine the sequence and thus
peptide/protein identities of candidate markers.
V U m c M e d i c a l O n c o l o g y – A n n u a l R e p or t 2 0 0 6
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Because this method yields identified proteins,
it can be more easily coupled to future antibodybased screening.
C. R esearch proteomics
Proteomics applied to model systems (in cell
culture or animal studies) is well established
and can solve a range of questions related
to protein expression levels, composition of
protein complexes, proteolytic processing,
post-translational modifications as well as (subcellular) localization and secretion into the
medium. Therefore these types of projects have
been started in 2006 immediately after the
required instruments were up and running.
42
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PhD theses 2006
PhD
Würdinger T. – The development of tumorselective Coronaviruses as anti-cancer agents.
Universiteit Utrecht. Promotor: prof.dr. P.J.M.
Rottier. Co-promotores: dr. W.R. Gerritsen,
dr. V.W. van Beusechem, dr. M.H.Verheije.
PrintPartners Ipskamp B.V., Enschede, 2006: 186
pp. Date: 01-02-2006.
Vuylsteke R.J.C.L.M. – Clinical and
immunological aspects of the sentinel lymph
node procedure in melanoma. Vrije Universiteit
Amsterdam. Promotores: prof.dr. P.A.M. van
Leeuwen, prof.dr. S.Meijer, prof.dr.R.J. Scheper.
Co-promotor: dr. T.D. de Gruijl. Febodruk BV,
Enschede, 2006; ISBN 90 8659 0144 4: 159 pp.
Date: 29-09-2006.
theses
2006
Vischioni B. – Expression and subcellular
localization of inhibitor of apoptosis (IAP)
proteins in human tissues: mechanisms
and clinical implications. Vrije Universiteit
Amsterdam. Promoto­r: prof. dr. G. Giaccone.
Co-promotor: dr. JA. Rodriguez. PrintPartners
Ipskamp B.V., Enschede, 2006: 192 pp. Date:
14-11-2006.
Witlox M.A. – Gene therapy of osteosarcoma.
Preclinical studies with adeoviral vectors
and adenoviral oncolysis. Vrije Universiteit
Amsterdam. Promotor: prof. dr. P.I.J.M.
Wuisman. Co-promotores: dr. V.W. van
Beusechem, dr. W.R. Gerritsen. Haveka B.V.,
2006. 161 pp. Date: 15-12-2006.
Dr. Barbara Vischioni with her paranymphs; Laudatio
by prof. Giaccone
Janmaat M.L. – Targeting ErbB receptors
as anticancer therapy: Factors of sensitivity
and resistance. Vrije Universiteit Amsterdam.
Promoto­r: prof. dr. G. Giaccone. Co-promotor:
dr. JA. Rodriguez. PrintPartners Ipskamp B.V.,
Enschede, 2006: 155 pp. Date: 11-10-2006.
43
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Scientific publicatons
Scientific
Abou El Hassan MA, Braam SR, Kruyt FA. A
real-time RT-PCR assay for the quantitative
determination of adenoviral gene expression in
tumor cells. J Virol Methods 2006; 133: 53-61.
Abou El Hassan MA, Braam SR, Kruyt FA.
Paclitaxel and vincristine potentiate adenoviral
oncolysis that is associated with cell cycle and
apoptosis modulation, whereas they differentially
affect the viral life cycle in non-small-cell lung
cancer cells. Cancer Gene Ther 2006; 13:1105-1114.
Achille M, Gallegos-Ruiz M, Giaccone G, Soria
JC. Response to erlotinib in first-line treatment
of non-small-cell lung cancer in a white male
smoker with squamous-cell histology. Clin Lung
Cancer 2006; 8: 214-216.
Ackland SP, Clarke SJ, Beale P, Peters GJ.
Thymidylate synthase inhibitors. Update Cancer
Ther. 2006; 1: 403-427.
Adema AD, Hubeek I, Zuurbier L, Floor K,
Albertioni F, Kaspers GJL, Peters GJ. Cellular
resistance against troxacitabine in human
cell lines and pediatric patient acute myeloid
leukemia blast cells. Nucleosides Nucleotides
Nucleic Acids 2006; 25: 981-986.
44
p u b l i ca t o n s
Bergman AM, Peters GJ. Gemcitabine.
mechanism of action and resistance. From:
Cancer drug discovery and development:
deoxynucleoside analogs in cancer therapy.
Edited by: G.J. Peters. Humana Press Inc.,
Totowa, NJ, USA. 2006; pp. 225-251.
Biesma B, Manegold C, Smit HJ, Willems L,
Legrand C, Passioukov A, van Meerbeeck JP,
Giaccone G; for the EORTC Lung Cancer
Group. Docetaxel and cisplatin as induction
chemotherapy in patients with pathologicallyproven stage IIIA N2 non-small cell lung cancer:
a phase II study of the European organization
for research and treatment of cancer (EORTC
08984). Eur J Cancer 2006; 42: 1399-1406.
Bijman MNA, Van Nieuw Amerongen GP,
Laurens N, Van Hinsbergh VWM, Boven
E. Microtubule-targeting agents inhibit
angiogenesis at subtoxic concentrations, a
process associated with inhibition of Rac1 and
Cdc42 activity and changes in the endothelial
cytoskeleton. Clin Cancer Res 2006; 5:2348-2357.
Boddaert MS, Gerritsen WR, Pinedo HM.
On our way to targeted therapy for cachexia in
cancer? Curr Opin Oncol 2006; 18: 335-340.
Bontkes HJ, Ruizendaal JJ, Kramer D, Santegoets
SJ, Scheper RJ, de Gruijl TD, Meijer CJ,
Hooijberg E. Constitutively active STAT5b
induces cytokine-independent growth of the
acute myeloid leukemia-derived MUTZ-3 cell line
and accelerates its differentiation into mature
dendritic cells. J Immunother 2006; 29: 188-200.
Bottomley A, Gaafar R, Manegold C, Burgers
S, Coens C, Legrand C, Vincent M, Giaccone
G, van Meerbeeck J; EORTC Lung-Cancer
Group; Natl Cancer Inst, Canada. Short-term
treatment-related symptoms and quality of life:
results from an international randomized phase
III study of cisplatin with or without raltitrexed
in patients with malignant pleural mesothelioma:
an EORTC Lung-Cancer Group and National
Cancer Institute, Canada, Intergroup Study. J
Clin Oncol 2006; 24: 1435-1442 (Erratum in: J
Clin Oncol 2006; 24: 2601).
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Bröker LE, de Vos FY, van Groeningen CJ,
Kuenen BC, Gall HE, Woo MH, Voi M, Gietema
JA, de Vries EG, Giaccone G. Phase I trial with
BMS-275183, a novel oral taxane with promising
antitumor activity. Clin Cancer Res 2006; 12:
1760-1767.
Broxterman HJ. Circulerende endotheelcellen bij
de kankerpatiënt. Angiogenese Journaal 2006; 1,
no. 2: 1-5.
Checinska A, Giaccone G, Hoogeland B,
Ferreira CG, Rodriguez JA, Kruyt FA. TUCAN
/CARDINAL/CARD8 and apoptosis resistance
in non-small cell lung cancer cells. BMC Cancer
2006; 6: 166.
Checinska A, Oudejans JJ, Span SW, Rodriguez
JA, Kruyt FA, Giaccone G. The expression of
TUCAN, an inhibitor of apoptosis protein, in
patients with advanced non-small cell lung cancer
treated with chemotherapy. Anticancer Res
2006; 26: 3819-3824.
De Bree R, van der Valk P, Kuik DJ, van Diest PJ,
Doornaert P, Buter J, Eerenstein SE, Langendijk
JA, van der Waal I, Leemans CR. Prognostic
factors in adult soft tissue sarcomas of the head
and neck: a single-centre experience. Oral Oncol
2006; 42: 703-709.
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De Bruin M, Temmink OH, Hoekman K,
Pinedo HM, Peters GJ. Role of platelet derived
endothelial cell growth factor / thymidine
phosphorylase in health and disease. Cancer
Ther 2006; 4: 99-124.
De Gruijl TD, Ophorst OJ, Goudsmit J, Verhaagh S,
Lougheed SM, Radosevic K, Havenga MJ, Scheper
RJ. Intradermal delivery of adenoviral type-35
vectors leads to high efficiency transduction of
mature, CD8+ T cell-stimulating skin-emigrated
dendritic cells. J Immunol 2006; 177: 2208-2215.
De Gruijl TD, Sombroek CC, Lougheed SM,
Oosterhoff D, Buter J, van den Eertwegh AJ,
Scheper RJ, Pinedo HM. A postmigrational
switch among skin-derived dendritic cells to a
macrophage-like phenotype is predetermined by
the intracutaneous cytokine balance. J Immunol
2006; 176: 7232-7242.
Efficace F, Bottomley A, Smit EF, Lianes P,
Legrand C, Debruyne C, Schramel F, Smit HJ,
Gaafar R, Biesma B, Manegold C, Coens C,
Giaccone G, van Meerbeeck J; the EORTC
Lung Cancer Group and Quality of Lif Unit. Is
a patient’s self-reported health-related quality of
life a prognostic factor for survival in non-smallcell lung cancer patients? A multivariate analysis
of prognostic factors of EORTC study 08975.
Ann Oncol 2006; 17: 1698-1704.
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Fijneman, RJ. Paving the road towards noninvasive molecular imaging. Cell Oncol 2006; 28:
125-126.
Fontijn D, Duyndam MC, van Berkel MP,
Yuana Y, Shapiro LH, Pinedo HM, Broxterman
HJ, Boven E. CD13/aminopeptidase N
overexpression by basic fibroblast growth factor
enhanced invasiveness of 1F6 human melanoma
cells. Br J Cancer 2006; 94: 1627-1636.
Giaccone G, Gallegos-Ruiz M. Le Chevalier T,
Thatcher N, Smit EF, Rodriguez JA, Janne P,
Oulid-Aissa D, Soria J-C. Erlotinib for frontline
treatment of advanced non-small cell lung
cancer: a phase II study. Clin Cancer Res 2006;
12: 6049-6055.
Giaccone G, Wilmink H, Paul MA, van der Valk
P. Systemic treatment of malignant thymoma: a
decade experience at a single institution. Am J
Clin Oncol 2006; 29: 336-344.
Graat HCA, Witlox MA, Schagen FHE, Kaspers
GJL, Helder MN, Bras J, Schaap GR, Gerritsen
WR, Wuisman PIJM, Van Beusechem VW.
Different susceptibility of osteosarcoma cell lines
and primary cells to treatment with oncolytic
adenovirus and doxorubicin or cisplatin. Br J
Cancer 2006; 94: 1837-1844.
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Haanen JB, Baars A, Gomez R, Weder P, Smits
M, de Gruijl TD, von Blomberg BM, Bloemena
E, Scheper RJ, van Ham SM, Pinedo HM, van
den Eertwegh AJ. Melanoma-specific tumorinfiltrating lymphocytes but not circulating
melanoma-specific T cells may predict survival
in resected advanced-stage melanoma patients.
Cancer Immunol Immunother 2006; 55: 451-458.
Haveman J, Sigmond J, Van Bree C, Franken
NA, Koedooder C, Peters GJ. Time course of
enhanced activity of deoxycytidine kinase and
thymidine kinase 1 and 2 in cultured human
squamous lung carcinoma cells, SW-1573, induced
by gamma-irradiation. Oncol Rep 2006; 16: 901905.
Honeywell R, van Groeningen CJ, Laan AC,
Strocchi E, Ruiter R, Giaccone G, Peters GJ.
Analysis of deoxycytidine accumulation in
gemcitabine treated patients. Nucleosides
Nucleotides Nucleic Acids 2006; 25: 1225-1232.
Hooijberg JH, De Vries NA, Kaspers GJL,
Pieters R, Jansen G, Peters GJ. Multidrug
resistance proteins and folate supplementation:
therapeutic implications for antifolates and other
classes of drugs in cancer treatment. Cancer
Chemother Pharmacol 2006; 58: 1-12.
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Hornberg JJ, Bruggeman FJ, Westerhoff HV,
Lankelma J. Cancer: a Systems Biology disease.
Biosystems 2006; 83: 81-90.
Hornberg JJ, Dekker H, Peters PH, Langerak P,
Lankelma J, van Zoelen EJ. Epidermal growth
factor receptor-induced activator protein 1
activity controls density-dependent growth
inhibition in normal rat kidney fibroblasts. Mol
Biotechnol 2006; 34: 101-108.
Hubeek I, Kaspers GJL, Ossenkoppele GJ,
Peters GJ. Cytosine arabinoside. Metabolism,
mechanisms of resistance and clinical
pharmacology. From: Cancer drug discovery and
development: deoxy-nucleoside analogs in cancer
therapy. Edited by: G.J. Peters. Humana Press
Inc., Totowa, NJ, USA. 2006; pp. 119-151.
Hubeek I, Peters GJ, Broekhuizen AJF, Zwaan
ChM,Kaaijk P, Van Wering ER, Gibson BES,
Creutzig U, Janka-Schaub GE, Den Boer ML,
Pieters R, Kaspers GJL. In vitro sensitivity and
cross-resistance to deoxynucleoside analogs in
childhood acute leukemia. Haematologica 2006;
91: 17-23.
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Janmaat ML, Gallegos-Ruiz M, Rodriguez
JA, Meijer GA, Vervenne WL, Richel DJ, Van
Groeningen C, Giaccone G. Predictive factors
for outcome in a phase II study of gefitinib in
second-line treatment of advanced esophageal
cancer patients. J Clin Oncol 2006; 24: 1612-1619.
Janmaat ML, Rodriguez JA, Gallegos-Ruiz M,
Kruyt FAE, Giaccone G. Enhanced cytotoxicity
induced by gefitinib and specific inhibitors of the
Ras or phosphatidyl inositol-3-kinase pathways in
non-small cell lung cancer cells. Int J Cancer 2006;
118: 209-214.
Janmaat ML, Rodriguez JA, Giaccone G.
Correspondence in reply to: Personeni N.,
Epidermal growth factor receptor gene copy
number in esophageal cancer and outcome
prediction to gefitinib: does intratumoral
heterogeneity matter? J Clin Oncol 2006; 24:
5466-5467.
Jellema AP, Slotman BJ, Muller MJ, Leemans
CR, Smeele LE, Hoekman K, Aaronson NK,
Langendijk JA. Radiotherapy alone, versus
radiotherapy with amifostine 3 times weekly,
versus radiotherapy with amifostine 5 times
weekly: A prospective randomized study in
squamous cell head and neck cancer. Cancer
2006; 107: 544-553.
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Jiménez CR. Proteomics in de Oncologie.
In: Kanker, tijdschrift van de Nederlandse
Vereniging voor Oncologie; april 2006.
Kramer GW, Legrand CL, van Schil P, Uitterhove
L, Smit EF, Schramel F, Biesma B, Tjan-Heijnen
V, van Zandwijk N, Splinter T, Giaccone G,
van Meerbeeck JP; on behalf of EORTC-Lung
Cancer Group. Quality assurance of thoracic
radiotherapy in EORTC 08941: a randomised
trial of surgery versus thoracic radiotherapy in
patients with stage IIIA non-small-cell lung
cancer (NSCLC) after response to induction
chemotherapy. Eur J Cancer 2006; 42: 1391-1398.
Kroep JR, Peters, GJ, Nagourney RA. Clinical
activity of gemcitabine as a single agent and in
combination. From: Cancer drug discovery and
development: deoxynucleoside analogs in cancer
therapy. Edited by: G.J. Peters. Humana Press
Inc., Totowa, NJ, USA. 2006; pp. 253-287.
Kroep JR, Smit EF, Giaccone G, van den Born
K, Beijnen JH, van Groeningen CJ, van der
Vijgh WJ, Postmus PE, Pinedo HM, Peters
GJ. Pharmacology of the paclitaxel-cisplatin,
gemcitabine-cisplatin, and paclitaxel-gemcitabine
combinations in patients with advanced nonsmall cell lung cancer. Cancer Chemother
Pharmacol 2006; 58: 509-516.
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Lamfers MLM, Fulci G, Gianni D, Tang Y,
Kurozumi K, Kaur B, Moeniralm S, Saeki Y,
Carette JE, Weissleder R, Vandertop WP, Van
Beusechem VW, Dirven CMF, Chiocca EA.
Cyclophosphamide increases transgene expression
mediated by an oncolytic adenovirus in gliomabearing mice monitored by bioluminescence
imaging. Mol Ther 2006; 14: 779-788.
Lammertsma AA, Hoekstra CJ, Giaccone G,
Hoekstra OS. How should we analyse FDG PET
studies for monitoring tumour response? Eur J
Nucl Med Mol Imaging 2006; 33 (Suppl 13): 16-21.
Lens SMA, Rodriguez JA, Vader G, Span SW,
Giaccone G, Medema RH. Uncoupling the
central spindle-associated function of the
Chromosome Passenger Complex from its role at
centromeres. Mol Biol Cell 2006; 17: 1897-1909.
Mauritz R, Peters GJ. Pharmacogenetics of
colon cancer and potential implications for
5-fluorouracil-based chemotherapy. Current
Pharmacogenomics 2006; 4: 57-67.
Molenkamp BG, van Leeuwen PA, van den
Eertwegh AJ, Sluijter BJ, Scheper RJ, Meijer
S, de Gruijl TD. Immunomodulation of the
melanoma sentinel lymph node: a novel adjuvant
therapeutic option. Immunobiology 2006; 211:
651-661.
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Padrón JM, Peters GJ. Cytotoxicity of sphingoid
marine compound analogs in mono- and
multilayered solid tumor cell cultures. Invest
New Drugs 2006; 24: 195-202.
Pauwels B, Korst AEC, Pattyn GGO,
Lambrechts HAJ, Kamphuis JAE, De Pooter
CMJ, Peters GJ, Lardon F, Vermorken JB.
The relation between deoxycytidine kinase
activity and the radiosensitizing effect of
gemcitabine in eight different human tumor cell
lines. BMC Cancer 2006; 6: 142 (doi: 10.1186/­
1471-2407-6-142).
Peltenburg H, Schopenhauer R, van der Linden
MHM. The Family Support project: a unique
concept of family care within the hospital
setting. Psychooncology 2006; 15: S1-S478.
Peters GJ. Preface. From: Cancer drug discovery
and development: deoxynucleoside analogs in
cancer therapy. Edited by: G.J. Peters. Humana
Press Inc., Totowa, NJ, USA. 2006; pp. V-VII.
Peters GJ, Carrey EA, Sebésta I. Purine and
pyrimidine metabolism, a firm base for a
transformed society. Nucleosides Nucleotides
Nucleic Acids 2006; 25: 971-974.
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Peters GJ, van Moorsel CJ, Lakerveld B, Smid K,
Noordhuis P, Comijn EC, Weaver D, Willey JC,
Voorn D, van der Vijgh WJ, Pinedo HM. Effects
of gemcitabine on cis-platinum-DNA adduct
formation and repair in a panel of gemcitabine
and cisplatin-sensitive or -resistant human
ovarian cancer cell lines. Int J Oncol 2006;
28: 237-244.
Santegoets SJ, Schreurs MW, Masterson AJ,
Liu YP, Goletz S, Baumeister H, Kueter EW,
Lougheed SM, van den Eertwegh AJ, Scheper
RJ, Hooijberg E, de Gruijl TD. In vitro priming
of tumor-specific cytotoxic T lymphocytes using
allogeneic dendritic cells derived from the human
MUTZ-3 cell line. Cancer Immunol Immunother
2006; 55: 1480-1490.
Rodriguez JA, Giaccone G. Keynote comment:
Are large-scale cancer-genomics projects ready to
use? Lancet Oncol 2006; 7: 190-191.
Savonije JH, van Groeningen CJ, Wormhoudt
LW, Giaccone G. Early Intervention with
epoetin alfa during platinum-based
chemotherapy: an analysis of quality-of-life
results of a multicenter, randomized, controlled
trial compared with population normative data.
Oncologist 2006; 11: 197-205.
Rodriguez JA, Lens SMA, Span SW, Vader
G, Medema RH, Kruyt FAE, Giaccone G.
Subcellular localization and nucleocytoplasmic
transport of the chromosomal passenger proteins
before nuclear envelope breakdown. Oncogene
2006; 25: 4867-4879.
Santegoets SJ, Masterson AJ, van der Sluis PC,
Lougheed SM, Fluitsma DM, van den Eertwegh
AJ, Pinedo HM, Scheper RJ, de Gruijl TD.
A CD34(+) human cell line model of myeloid
dendritic cell differentiation: evidence for a
CD14(+)CD11b(+) Langerhans cell precursor. J
Leukoc Biol 2006; 80: 1337-1344.
48
Ne
n t e n t s
Savonije JH, van Groeningen CJ, Wormhoudt
LW, Giaccone G. Early intervention with epoetin
alfa during platinum-based chemotherapy:
an analysis of the results of a multicenter,
randomized, controlled trial based on initial
hemoglobin level. Oncologist 2006; 11: 206-216.
Savonije JH, van Groeningen CJ, van den Broek
WJ, Rentinck ME, Corstens FH, Gundy C,
Giaccone G, Marx JJ. Iron absorption during
epoetin alfa therapy for chemotherapy-associated
anaemia. Cancer Invest 2006; 24: 562-566.
x t
Schagen FHE, Wensveen FM, Carette JE,
Dermody TS, Gerritsen WR, Van Beusechem
VW. Genetic targeting of adenovirus vectors
using a reovirus σ1-based attachment protein.
Mol Ther 2006; 13: 997-1005.
Sigmond J, Haveman J, Castro Kreder N,
Loves WJ, Van Bree C, Franken NA, Peters GJ.
Enhanced activity of deoxycytidine kinase after
pulsed low dose rate and single dose gammairradiation. Nucleosides Nucleotides Nucleic
Acids 2006; 25: 1177-1180.
Smid K, Bergman AM, Eijk PP, Veerman G,
Ruiz van Haperen VWT, Van den IJssel P, Ylstra
B, Peters GJ. Micro-array analysis of resistance
for gemcitabine results in increased expression of
ribonucleotide reductase subunits. Nucleosides
Nucleotides Nucleic Acids, 2006; 25: 1001-1007.
Smorenburg CH, Peters GJ, van Groeningen
CJ, Noordhuis P, Smid K, van Riel AM,
Dercksen W, Pinedo HM, Giaccone G. Phase
II study of tailored chemotherapy for advanced
colorectal cancer with either 5-fluouracil and
leucovorin or oxaliplatin and irinotecan based
on the expression of thymidylate synthase and
dihydropyrimidine dehydrogenase. Ann Oncol
2006; 17: 35-42.
V U m c M e d i c a l O n c o l o g y – A n n u a l R e p or t 2 0 0 6
Close
Pr
Co
e v i ou s
Temmink OH, De Bruin M, Laan AC, Turksma
AW, Cricca S, Masterson AJ, Noordhuis P, Peters
GJ. The role of thymidine phosphorylase and
uridine phosphorylase in (fluoro) pyrimidine
metabolism in peripheral blood mononuclear
cells. Int J Biochem Cell Biol 2006; 38: 1759-1765.
Temmink OH, Hoogeland MFM, Fukushima M,
Peters GJ.Low folate conditions may enhance the
interaction of trifluorothymidine with antifolates
in colon cancer cells. Cancer Chemother
Pharmacol 2006; 57: 171-179.
Tijink BM, Buter J, de Bree R, Giaccone G, Lang
MS, Staab A, Leemans CR, van Dongen GA. A
phase I dose escalation study with anti-CD44v6
bivatuzumab mertansine in patients with
incurable squamous cell carcinoma of the head
and neck or esophagus. Clin Cancer Res 2006;
12(20 Pt 1): 6064-6072.
Van der Linden MHM, Ruijter S, Verdonck-de
Leeuw IM, Kleverlaan N. Children of parents
with cancer, an exploratory study of the Dutch
website ‘Cancer Phantoms’. Psychooncology
2006; 15: S46-47.
Van der Vliet HJ, van Vonderen MG, Molling JW,
Bontkes HJ, Reijm M, Reiss P, van Agtmael MA,
Danner SA, van den Eertwegh AJ, von Blomberg
BM, Scheper RJ. Cutting edge: Rapid recovery
49
Ne
n t e n t s
of NKT cells upon institution of highly active
antiretroviral therapy for HIV-1 infection.
J Immunol 2006; 177: 5775-5778.
Van de Ven R, de Jong MC, Reurs AW,
Schoonderwoerd AJ, Jansen G, Hooijberg JH,
Scheffer GL, de Gruijl TD, Scheper RJ. Dendritic
cells require multidrug resistance protein 1
(ABCC1) transporter activity for differentiation.
J Immunol 2006; 176: 5191-5198.
Van Heijst JW, Niessen HW, Musters RJ, van
Hinsbergh VW, Hoekman K, Schalkwijk CG.
Argpyrimidine-modified Heat shock protein 27
in human non-small cell lung cancer: a possible
mechanism for evasion of apoptosis. Cancer Lett
2006; 241: 309-319.
Van Hinsbergh VWM, Engelse MA, Quax
PHA. Pericellular proteases in angiogenesis and
vasculogenesis. Arterioscler Thromb Vasc Biol
2006; 26: 716-728.
Verdonck-de Leeuw IM, Borggreven PA,
Eerenstein SEJ, van der Linden MHM, Rinkel
RNPM, de Bree R, Leemans CR. Psychosociale
gevolgen en functionele gevolgen van hoofdhalskanker. Nederlands Tijdschrift voor
Oncologie 2006; 3: 185-191.
x t
Verdonck-de Leeuw IM, Eerenstein SEJ,
van der Linden MHM, Kuik DJ, de Bree R,
Leemans CR. Distress in spouses and patients
with head and neck cancer. Psychooncology
2006; 15: S1-S478.
Vergeer MR, Doornaert P, Leemans CR,
Buter J, Slotman BJ, Langendijk JA. Control
of nodal metastases in squamous cell head and
neck cancer treated by radiation therapy or
chemoradiation. Radiother Oncol 2006; 79: 3944.
Verheije MH, Würdinger T, Van Beusechem VW,
De Haan CAM, Gerritsen WR, Rottier PJM.
Redirecting coronavirus to a non-native receptor
through a virus-encoded targeting adapter. J Virol
2006; 80: 1250-1260.
Vischioni B, Oudejans JJ, Vos W, Rodriguez JA,
Giaccone G. Frequent overexpression of aurora
B kinase, a novel drug target, in non-small cell
lung carcinoma patients. Mol Cancer Ther 2006;
5: 2905-2913.
Vischioni B, van der Valk P, Span SW, Kruyt
FAE, Rodriguez JA, Giaccone G. Expression and
localization of inhibitor of apoptosis proteins in
normal human tissues. Human Pathol 2006; 37:
78-86.
V U m c M e d i c a l O n c o l o g y – A n n u a l R e p or t 2 0 0 6
Close
Pr
Co
e v i ou s
n t e n t s
Voortman J, Giaccone G. Severe reversible
cardiac failure after bortezomib treatment
combined with chemotherapy in a non-small cell
lung cancer patient: a case report. BMC Cancer
2006; 6: 129.
Vuylsteke RJ, Molenkamp BG, van Leeuwen
PA, Meijer S, Wijnands PG, Haanen JB, Scheper
RJ, de Gruijl TD. Tumor-specific CD8+ T
cell reactivity in the sentinel lymph node of
GM-CSF-treated stage I melanoma patients
is associated with high myeloid dendritic cell
content. Clin Cancer Res 2006; 12: 2826-2833.
Würdinger T, Verheije MH., Van der Aa LM,
Bosch BJ, De Haan CAM, Van Beusechem
VW, Gerritsen WR, Rottier PJM. Antibodymediated targeting of coronaviral vectors to the
Fc receptor on acute myeloid leukemia cells.
Leukemia 2006; 20: 2182-2184.
Zucali PA, Giaccone G. Biology and management
of malignant pleural mesothelioma. Eur J Cancer
2006; 42: 2706-2714.
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V U m c M e d i c a l O n c o l o g y – A n n u a l R e p or t 2 0 0 6
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