Download Talk Title: Regulation of dendritic cell development at steady

Talk Title: Regulation of dendritic cell development at steady-­‐state and inflammation Speaker: Chien-­‐Kuo LEE During infections and inflammation, plasmacytoid dendritic cells (pDCs) are the most potent type I interferon (IFN-­‐I)-­‐producing cells. However, the developmental origin of pDCs and the signals dictating pDC generation at steady-­‐state and inflammation remain incompletely understood. Here, we report a synergistic role for IFN-­‐I and Flt3 ligand (FL) in pDC development from common lymphoid progenitors (CLPs). Both conventional DCs (cDCs) and pDCs were generated from CLPs in response to FL, whereas pDC generation required higher concentrations of FL and concurrent IFN-­‐I signaling. An absence of IFN-­‐I receptor, impairment of IFN-­‐I signaling, or neutralization of IFNAR significantly impeded pDC development from CLPs. This is, in part, due to reduced expression of Flt3 on CLPs lacking IFNAR1. Restoration of Flt3 expression on IFNAR1KO CLPs rescued FL-­‐dependent pDC development from CLPs. Furthermore, FL induced IFN-­‐I expression in CLPs, which in turn increased Flt3 up-­‐regulation that facilitated survival and proliferation of CLPs, as well as their differentiation into pDCs. We also investigate the effect of inflammation on DC development from CLPs. Interestingly, CpG and other TLR stimulation altered the FL-­‐dependent developmental program of DCs from CLPs. The inflammation signals changed the developmental process from pro-­‐pDC to pro-­‐cDC. The effect was intrinsic to TLR signal-­‐mediated pathways. Collectively, these results define a critical role for the FL/IFN-­‐I/Flt3 axis in pDC differentiation at steady-­‐state and TLR signaling in cDC differentiation during inflammation. Talk title: Dysregulation of MDA5-­‐dependent signaling causes autoimmune disorder Speaker: Takashi FUJITA MDA5 is an essential intracellular sensor for several viruses, including picornaviruses, and elicits antiviral interferon (IFN) responses by recognizing viral dsRNAs. Once MDA5 senses replicating viruses, it triggers signal to activate antiviral genes including those of type I and III IFN. Activation of IFN system is critical as antiviral innate immunity and promotes activation of acquired immunity. These immune responses orchestrate eradication of infecting viruses. On the other hand, MDA5 has been implicated in autoimmunity. The mechanisms of how MDA5 contribute to autoimmunity remain unclear. Here we provide direct evidence that dysregulation of MDA5 caused autoimmune disorders. We established a mutant mouse line bearing MDA5 mutation by ENU mutagenesis, which spontaneously developed lupus-­‐like autoimmune symptoms without viral infection. Inflammation was dependent on an adaptor molecule, IPS-­‐1, indicating the importance of MDA5-­‐signaling. In addition, intercrossing the mutant mice with type I IFN receptor-­‐deficient mice ameliorated clinical manifestations. This MDA5 mutant could activate signaling in the absence of its ligand, but was paradoxically defective for ligand-­‐ and virus-­‐induced signaling, suggesting that the mutation induces a conformational change in MDA5. These findings provide insight into the association between disorders of the innate immune system and autoimmunity. Talk title: Translational studies on clinical immunological diseases Speaker: Bor-­‐Luen CHIANG In the past 10 years, our team has focused on developing diagnostic and therapeutic approaches for immunological diseases. To further explore the possible therapeutic approaches, understanding the immune regulatory mechanisms become extremely critical. Numerous studies performed with asthma animal models have shown increased T helper 2 (TH2) cytokine levels and decreased T helper 1 (TH1) cytokine levels within the affected airways. Moreover, TH2 cells together with other inflammatory cells, such as eosinophils, mast cells and B cells play a major role in the initiation and pathological development of asthma. In the past several years, our team has applied cytokine gene and also siRNA for the regulation of immune balance in allergic immune responses. In addition, we have also applied siRNA for the treatment of the tumor. To apply the approaches of immune regulation for immunologic diseases, we have investigated the mechanisms of mucosal tolerance. Our previous study with mite Derp2 allergen transgenic plant demonstrated that proteins derived from transgenic plant could decrease Derp2-­‐specific IgE level and also alleviate airway inflammation. Further studies suggested that mucosal B cells play a critical role in the oral tolerance of immunological diseases. Our recent studies also showed regulatory T cells (Treg/B cells) induced by mucosal B cells could exert suppressive effect on T cells function and also alleviate the airway inflammation in animal model of asthma. We have also applied these Treg/B cells for the treatment of animal model of collagen-­‐induced arthritis. In addition, we have also explored the novel diagnostic method to define the allergic shiner and also the biomarkers for Henoch-­‐Scholein purpura, a pediatric vasculitic disease. All these results might be used for the future application of immunologic diseases. In summary, our team aim to develop the novel both diagnostic and therapeutic approaches for immunologic diseases. Talk title: Exposure of phosphatidylserine, and engulfment of apoptotic cells by macrophages Speaker: Shigekazu NAGATA Apoptotic cells are swiftly engulfed and digested by macrophages. If this process does not occurs properly, materials released from dead cells activate the immune system, leading to systemic lupus erythematosus (SLE)-­‐type autoimmune disease. Phospholipids are most abundant lipids in plasma membranes, and are asymmetrically distributed between inner and outer leaflets. That is, phosphatidylserine (PtdSer) and phosphatidylethanolamine (PtdEtn) are exclusively localized in the inner leaflet, while phosphatidylcholine (PtdCho) and sphingomyelin (SM) are mainly on the outer leaflet. The asymmetrical distribution of phospholipids is maintained by an ATP-­‐dependent phospholipid translocase or flippase. When cells undergo apoptosis, or platelets are activated, the asymmetrical distribution of phospholipids is disrupted by scramblase, leading to PtdSer-­‐exposure. The PtdSer exposed on the dead cell surface is recognized by macrophages as an “eat me” signal, while PtdSer on the activated plates provide the scaffold for blood clotting factors. The molecular identity of flippase and scramblase had been elusive for a long time, but we recently identified two membrane proteins (TMEM16F and Xkr8) as phospholipid scramblases, and a pair of membrane proteins (ATP11C and CDC50A) as a flippase. TMEM16F carries 8 transmembrane regions, and requires Ca2+ to mediate phospholipid scrambling. It plays an essential role in the PtdSer-­‐exposure in activated platelets, and patients of Scott Syndrome who suffer bleeding disorder carry a loss-­‐of function mutation in TMEM16F gene. Xkr8 is a protein carrying 6 transmembrane regions, and caspase 3 and 7, cysteine proteases that are activated during apoptosis, cleave off the C-­‐terminal tail of Xkr8 to promote its scramblase activity. ATP11C is a P4-­‐type ATPases at plasma membrane, and CDC50A works as a chaperone to transport ATP11C from endoplasmic reticulum to plasma membranes. ATP11C has the ability to translocate PtdSer from outer to inner leaflets of plasma membranes in an ATP-­‐dependent manner. When cells undergo apoptosis, ATP11C is inactivated by caspase-­‐mediated cleavage. The cells that completely lack the flippase activity (i.e. CDC50A-­‐null cells) constitutively expose PtdSer, and are engulfed by macrophages. These results indicate that during apoptosis, caspases activate phospholipid scramblase, and inactivate flippase, and PtdSer is necessary and sufficient as an “eat me” signal to be recognized by macrophages. References Nagata, S., Hanayama, R., and Kawane, K. (2010). Autoimmunity and the Clearance of Dead Cells. Cell 140, 619-­‐630. Suzuki, J., Umeda, M., Sims, P.J., and Nagata, S. (2010). Calcium-­‐dependent phospholipid scrambling by TMEM16F. Nature 468, 834-­‐838. Suzuki, J., Denning, D.P., Imanishi, E., Horvitz, H.R., and Nagata, S. (2013). Xk-­‐related protein 8 and CED-­‐8 promote phosphatidylserine exposure in apoptotic cells. Science 341, 403-­‐406. Segawa, K., Kurata, S., Yanagihashi, Y., Brummelkamp, T., Matsuda, F., and Nagata, S. (2014) Caspase-­‐mediated cleavage of phospholipid flippase for apoptotic phosphatidylserine exposure. Science 344, 1164-­‐1168. Talk Title: Activation of topoisomerase II-­‐mediated DNA cleavage and mutagenesis during
inflammation underlies cancer development Speaker: Tsai-­‐Kun LI Tissue inflammation such as gastritis, hepatitis and colitis (e.g. induced by infection of bacteria or by chemicals) is recognized a risk factor for human cancers at various sites, where reactive nitrogen and oxygen species (RNOS, e.g. nitric oxide NO) and RNOS-­‐mediated macromolecule damages contribute to the carcinogenic process in inflamed tissues. Recently, chronic inflammation and genomic instability have been defined as the two cancer enabling characteristics during tumorigenesis. However, the mechanism(s) underlying the carcinogenic action of RNOS and the exact nature of this relationship remain unknown. In this regard, two mouse models, namely the chemical-­‐induced ulcerative colitis in colon (mimicking inflammatory bowl diseases, IBD) and the 2-­‐stage skin carcinogenesis model, were employed. We showed here, for the 1st time, that expression of inducible NO synthase (iNOS/NOS2) is responsible for activation of DNA damage response (DDR, e.g. γ-­‐H2AX expression), but minimally contributes to the induction of inflammatory response, in colon ulcerative colitis. We also showed that NO produced during either chronic inflammation or treatment of a NO releaser GSNO induced DNA breaks and mutagenesis in cells, and skin melanoma formation on mice. Pharmacological inhibition of topoisomerase II (TOP2α and TOP2β) isozymes by ICRF-­‐193 abolished DDR activation in colitis tissues. Generation of DNA damage, mutagenesis, and melanoma was mediated primarily by the TOP2β isozyme, which could be also inhibited by ICRF-­‐193. Similarly, we have showed that TOP2β-­‐mediated breaks and mutagenesis contribute to skin melanoma formation upon exposure of carcinogenic metal cadmium. Our study thus provides a novel insight into the mechanism by which the activated TOP2β-­‐mediated breakage links pro-­‐cancer characteristics to the genesis of genetic abnormalities that subsequently expediting the acquisition of cancer propensities and fostering their pathological functions. Our results also suggest that NO scavengers, iNOS and TOP2β inhibitors are potentially cancer-­‐preventive, especially for those related to inflammatory bowl diseases and/or due to exposure of the carcinogenic metals. Interestingly, out recent structure-­‐based and rationally designed compounds have started to reveal isozyme specificity against TOP2β. Talk title: AID-­‐regulated Topoisomerase1 is the DNA cleaving enzyme during immunoglobulin diversification Speaker: Maki KOBAYASHI The genetic alterations of the immunoglobulin (Ig) gene including class switch recombination (CSR), somatic hypermutation (SHM) and gene conversion (GC) amplify antibody diversity so that the immune system protects individuals from infection of the micro-­‐organisms. CSR, SHM and GC are dependent on activation-­‐induced cytidine deaminase (AID) and initiated by single-­‐strand DNA cleavage at the repeated sequences which are prone to form non-­‐B DNA structure when actively transcribed. Abberant AID activation causes genomic instability by multiple mutations and/or translocation in cancer-­‐related genes and eventually induces malignancy. Although AID's enzymatic activity is deamination of cytidine, one of the essential AID’s function is triggering of multiple DNA breaks in the relatively limited genomic region. The enzyme which actually cuts DNA, working under the control of AID, was not clear for many years after AID discovery. We had found that AID suppresses topoisomerase I (Top1) translation to decrease Top1 protein amount, which in turn induces non-­‐B DNA structure of the repeated DNA sequences. Top1 nicks non-­‐B DNA structure in Ig and the other target loci of AID. We obtained the following evidence, 1) Top1 specific inhibitor camptothecin suppressed CSR and SHM 2) Top1 knockdown or haploinsufficiency in the heterozygotes have augmented CSR and SHM. Conversely, Top1 overexpression reduces SHM. Interestingly, this Top1 involvement in transcription-­‐dependent DNA recombination at repeat-­‐rich DNA is shared by transcription-­‐associated mutagenesis (TAM) in yeast and triplet-­‐repeat contraction/expansion in triplet repeat diseases like Huntington’s chorea or Freidreich’s ataxia. We have investigated how AID regulates translation of Top1. We provide evidence to support that AID might edits some miRNA's precursor to promote the recruitment of RISC complex to Top1 mRNA in cooperation with an RNA-­‐binding protein, HuR. Recent progress in understanding the mechanism of DNA cleavage by Top1 will be addressed in this talk. Talk Title: Telomerase activation in stem cells and in yeast Speaker: Shu-­‐Chun TENG Telomeres are dynamic DNA-­‐protein complexes that protect the ends of linear chromosomes. Most telomeric DNA is synthesized by the enzyme telomerase. While most somatic cells do not express telomerase and therefore have limited life span, cancer cells can bypass the crisis either through telomerase reactivation or through an alternative recombination pathway for telomere lengthening (ALT). We will discuss our recent findings of telomere replication on telomerase expression, telomerase activation, and ALT progression. The Krüppel-­‐like transcription factor 4 (KLF4) has been implicated in cancer formation and stem cell regulation. However, the function of KLF4 in tumorigenesis and stem cell regulation are poorly understood due to limited knowledge of its targets in these cells. We have revealed a surprising link between KLF4 and regulation of telomerase, which offers important insight into how KLF4 contributes to cancer formation and stem cell regulation. KLF4 directly activate expression of the human telomerase catalytic subunit, hTERT. Our findings demonstrate that hTERT is one of the major targets of KLF4 in cancer and stem cells to maintain long-­‐term proliferation potential. Moreover, we demonstrated that telomerase is mainly activated by Cdk1/Tel1/Mec1 on telomeric binding protein Cdc13 from late S to G2 phase of the cell cycle. Phenotypic analysis in vivo revealed that the mutations in the Cdc13 S/TP motifs phosphorylated by Cdk1/Mec1/Tel1 caused cell cycle delay and telomere shortening and these phenotypes could be partially restored by the replacement with a negative charge residue. Furthermore, the Cdk1-­‐mediated phosphorylation was required to promote the regular turnover of Cdc13. Hypernegatively charged domain of Cdc13 contributed by Cdk1, Tel1 and Mec1 may provide an optimal interface to recruit the potential positively charged domain near the amino acid 444 lysine residue of Est1 in the telomerase complex. Together these results demonstrate that Cdk1/Mec1/Tel1 phosphorylate the telomerase recruitment domain of Cdc13, thereby preserves optimal function and expression level of Cdc13 for precise telomere replication and cell cycle progression. Talk title: How telomerase is recruited to telomeres in human cells Speaker: Fuyuki ISHIKAWA Telomerase is a specialized reverse transcriptase that de novo synthesizes telomere DNA repeats. The reaction compensates the gradual telomere erosion caused by the end-­‐replication problem, thus enabling the germ cells and cancer cells to proliferate unlimitedly. As such, it has been expected that telomerase inactivation in cancer cells would limit the tumor growth. However, efforts to develop drugs targeting the catalysis of telomerase have been not successful. Another possibility to limit the telomere extension by telomerase is inhibiting the recruitment of telomerase to its substrate telomeres. It has been reported that the telomere protein TPP1 recruits telomerase in mammalian cells. We have found that TPP1 undergoes a cell cycle-­‐dependent phosphorylation. Abrogating the phosphorylation leads to faster telomere shortening. These results indicate that the phosphorylation reaction is a candidate target to prohibit telomerase reaction in cancer cells. Talk Title: Structure-­‐based development of selective type II topoisomerase-­‐targeting anticancer drugs Speaker: Nei-­‐Li CHAN Drugs targeting type II DNA topoisomerases (Top2s) are among the most widely prescribed chemotherapeutic agents for treating cancers. By interfering with the catalytic cycle of Top2, these drugs exert cell-­‐killing activity by promoting the formation of enzyme-­‐mediated DNA double-­‐strand breaks to initiate the cell death pathways. Despite their effectiveness, the prolonged administration of these drugs are known to cause serious side effects, including therapy-­‐related secondary leukemia and cardiotoxicity. Mounting evidences suggest that the undifferentiated drug-­‐targeting of both human Top2 isoforms, hTop2α and hTop2β, is likely the primary cause of adversity: while targeting of hTop2α is sufficient for killing cancer cells, the hTop2β-­‐induced DNA breaks and chromosome translocation events result in side effects. To overcome these problems, it would be clinically desirable to develop a hTop2α-­‐specific targeting agent (poison) to trigger cell-­‐killing and a hTop2β-­‐specific catalytic inhibitor to suppress side effects. Toward this goal, we have determined the high-­‐resolution crystal structures of clinically active anticancer drugs in complexes with DNA and both human Top2 isoforms. The identification of enzyme-­‐drug interactions has not only revealed the structural basis of drug action but also allowed a set of drug-­‐design guidelines to be formulated. To speed up the drug development process, we are actively pursuing structure-­‐based modification of FDA-­‐approved Top2-­‐targeting anticancer drugs to introduce isoform-­‐specific targeting activity. The progress of this ongoing effort will be presented during the meeting. Talk title: Structural biology of membrane proteins and drug discovery Speaker: So IWATA Membrane proteins are a supreme example where more effort in structural biology is needed. In spite of their abundance and importance, of the 100,000+ protein structures in the Protein Data Bank, only some 500 of these are for unique membrane proteins. Many major questions remain unanswered; these include the signal transduction mechanism of receptors, the gating mechanisms of ion channels, and the molecular transport mechanism of transporters. We are currently trying to establish a pipeline for mammalian membrane-­‐protein structures, particularly focusing on drug-­‐targets, by combining our expertizes on expression, crystallization and X-­‐ray data collection. We are also developing the data collection system, suitable for data collection from membrane protein crystals, at SACLA, a new Japanese X-­‐ray free electron laser facility. In my talk, I will present structures of drug-­‐target membrane proteins including GPCRs and the Rce1 membrane proteinase and discuss current status of membrane protein crystallography on drug-­‐targets and future prospects. Talk title: Design and synthesis of A neurotrophic and fibrillar β-­‐amyloid dissolution molecule for Alzheimer’s disease Speaker: Ji-­‐Wang CHERN Alzheimer's disease is a neurodegenerative disorder characterized by progressive neurite loss and fibrillar β-­‐amyloid deposition. None of the clinically approved anti-­‐Alzheimer’s agents has been reported on either these two pathological processes. Here, an alkyl alcohol-­‐substituted quinoline (J2326) was created by joining an 11-­‐carbon alkyl alcohol to 5-­‐chloro-­‐8-­‐methoxyquinoline, stimulates neurite outgrowth and dissociates β-­‐amyloid fibrils. It was observed that J2326 triggered ERK-­‐dependent neurite outgrowth and regrowth accompanied by enhanced synaptic function in vitro. In addition, J2326 attenuated β-­‐amyloid aggregation and stimulated the dissociation of pre-­‐existing aggregates. Besides, we also demonstrated that J2326 binds to β-­‐amyloid fibrils. Moreover, J2326 exhibits neuroprotective function associating with its anti-­‐oxidant and anti-­‐caspase effects. Animal studies showed that J2326-­‐stimulated regrowth of dystrophic neurites are pivotal for the improvement of memory in mice with fibrillar β-­‐amyloid-­‐induced lesions. J2326 has pluripotent effects on neuroprotection and neuritogenesis and stands for a novel compound for the development of anti-­‐Alzheimer’s agents. Talk title: Challenges to cure genetic diseases with RNA-­‐targeting chemical compounds Speaker: Masatoshi HAGIWARA Patients of congenital diseases have abnormalities in their chromosomes and/or genes. Therefore, it has been considered that drug treatments can serve to do little for these patients more than to patch over each symptom temporarily when it arises. Although we cannot normalize their chromosomes and genes with chemical drugs, we may be able to manipulate the amounts and patterns of mRNAs transcribed from patients DNAs with small chemicals. Based on this simple idea, we have looked for chemical compounds which can be applicable for congenital diseases and found INDY, TG003, and SRPIN340 are promising as clinical drugs for Down syndrome (DS), Duchenne muscular dystrophy (DMD), and Denys Drash Syndrome (DDS), respectively. Familial dysautonomia (FD), a hereditary sensory and autonomic neuropathy, is caused by mis-­‐splicing resulting from an intronic mutation in IKBKAP gene. FD would be treatable if we can develop “a splicing modulator” which promotes exon20 inclusion of IKBKAP and increases the expression of IKAP protein in FD patient cells. In order to find the modulator, we established splicing reporter assay with dual color (SPREAD) using a segment of human IKBKAP spanning from exon19 to exon21. SPREAD allows us to visualize the splicing in cells, and to identify RBM24 and RBM38 as the tissue-­‐specific modulators for exon20 inclusion of IKBKAP. This also enabled us to find a chemical compound RECTAS, which can rectify the aberrant IKBKAP splicing in FD patient fibroblasts. Our data implicate the mis-­‐splicing of IKBKAP in the reduced tRNA modification in FD patient and demonstrated that RECTAS could be the therapeutic drug. References 1. Amin EM, Hua J, Cheung MK, Ni L, Kase S, Ren-­‐nel ES, Gammons M, Nowak DG, Saleem MA, Hagiwara M, Schumacher VA, Harper SJ, Hinton D, Bates DO, Ladomery MR (2011) WT1 mutants reveal SRPK1 to be a downstream angiogenesis target by altering VEGF splicing. Cancer Cell 20(6):768-­‐780. 2. Nishida A, Kataoka N, Takeshima Y, Yagi M, Awano, H, Ota, M, Itoh K, Hagiwara M, and Matsuo M (2011) Chemical treatment enhances skipping of a mutated exon in the dystrophin gene. Nature Communications 2, 308. 3. Ogawa Y, Nonaka Y, Goto T, Ohnishi E, Hiramatsu T, Kii I, Yoshida M , Ikura T , Onogi H , Shibuya H , Hosoya T, Ito N, and Hagiwara M (2010) Development of a novel selective inhibitor of the Down syndrome-­‐related kinase Dyrk1A. Nature Communications.1 , 86. Talk title: Central role of inflammasome for the host response to bacterial infection Speaker: Masao MITSUYAMA In combating against pathogenic bacteria, the host mounts innate immune response at the early stage of infection as well as the antigen-­‐specific immune response at the later stage. Th1-­‐type immune response is indispensable for the complete elimination of multiplying bacteria, and several cytokines including IL-­‐1, IL-­‐12 and IL-­‐18 produced at the early stage of infection play a critical role in the induction of antigen-­‐specific T cell response. Among them, IL-­‐18 is an essential cytokine in mounting the Th1 response, and its maturation from pro-­‐IL-­‐18 into fully functional form requires the activation of caspase-­‐1. Listeria monocytogenes is a representative intracellular parasitic bacterium that induces a robust Il-­‐18 production followed by establishment of Th1-­‐dependent cell-­‐mediated immunity. Based on our own observation that listeriolysin O (LLO), a major virulence factor of this bacterium, is essential for the induction of both IL-­‐18 response and Th1-­‐dependent immunity in the infected host, we have constructed numbers of LLO mutant and have identified the domain responsible for caspase-­‐1 activation. This finding has clarified the reason why only LLO-­‐positive virulent strains of L. monocytogenes are capable for induction of Th1 response in infected host. In the process of caspase-­‐1 activation, inflammasome formation is indispensable. There have been reported two major pathways including NLRP3-­‐dependent and NLRC4-­‐dependent pathways. In our recent study, we have identified a novel pathway that involves AIM2 as the third major pathway of inflammasome formation resulting in an ASC-­‐dependent caspase-­‐1 activation. Regarding the detailed molecular mechanism of caspase-­‐1 activation, we have recently found that phosphorylation of Y144 of ASC is critical for inflammasome-­‐dependent caspase-­‐1 activation and also that ASC spec formation is highly dependent on Syk and JNK. These recent findings have proposed an idea that the presence of virulence factor in Listeria monocytogenes plays a central role in the induction of Th1-­‐depensent immune response through the formation of inflammasome and caspase-­‐1-­‐dependent maturation of IL-­‐18. Our recent findings of the host response against virulent Streptococcus pneumoniae regarding ASC and inflammasome will additionally be presented and discussed. Talk Title: Comparative effectiveness evaluation of medications using a national population-­‐based healthcare environment Speaker: K. Arnold CHAN While the term translational medicine has become very popular in recent years, most of the time the focus is on developing promising molecules from test tube to regulatory approval. However, according to certain definition, the ultimate goal of translational medicine is to have effective health care products widely adopted in clinical practice, resulting in tangible improvement of global health. From this wider perspective of translational medicine, comparative effectiveness research is a critical component, which is at least as important as the pivotal phase III clinical trials. In this presentation, the relevant methods and data sources for comparativeness will be reviewed. Real world examples of evaluation for treatment regimens of viral hepatitis B and non-­‐small cell lung cancer and long-­‐term management of myocardial infarction will be presented to illustrate the important role of comparative effectiveness research. Talk title: Translational research in Kyoto University Hospital Speaker: Akira SHIMIZU To improve outcome of treatment or prevention from incurable diseases by creation and development of novel medical treatments and diagnoses is one of the big missions given to the Kyoto University Graduate School of Medicine and to Kyoto University Hospital. For this purpose, in April 2001, Kyoto University Hospital took a new strategy to convert the findings of basic medicine and biology efficiently into novel clinical diagnosis and treatment by establishing a center, named Translational Research Center (TRC), to promote such translational and synthetic research, especially to take proof of concept (POC) in humans by early phase (Phase I-­‐IIa) clinical trials. In 2012, Kyoto University Hospital was selected as one of Core Clinical Research Hospital (CCRH) by the Ministry of Health, Labor and Welfare. CCRH implies that besides conducting clinical trials and investigator-­‐initiated IND/IDE trials of international standards, it also is stands to be a base for medical devices network based hospital. In response to this designation, the Kyoto University Hospital Core Clinical Research Hospitals Initiative was assessed, and departments of TRC, Clinical Trial Control Center, EBM Research Center and Department of R&D and Corporate Integration was unified into one organization and make a fresh start from April 2013 as Institute for Advancement of Clinical and Translational Science (iACT). In order enable new therapies that are developed through clinical research proposals to be applied to humans, a wide range of operations is essential starting from the strategic development of research proposals to the clinical deployment and compilation of research results. The Department of Experimental Therapeutics, one of three departments in former TRC and now in iACT, organizes and conducts these comprehensive activities. This department has supported many such early exploratory clinical trials so far. Some examples of investigator initiated IND trials for POC done or now going on at Kyoto University Hospital will be presented. Talk title: The role of ultrasound in breast cancer screening and the development of automated whole breast ultrasound Speaker: Chiun-­‐Sheng HUANG The incidence of breast cancer has increased dramatically in Taiwan in the past twenty years. However, in contrast to incidence rates in Western countries, the incidence of breast cancer in Taiwan is considered to be among the lowest in the world. Although the incidence is low, breast cancer is the most common cancer in women in Taiwan. In addition, less than 10 % of breast cancer is non-­‐invasive and breast cancer is among the leading causes of cancer mortality in women with cancers, which indicates a need of breast cancer screening program in Taiwan. As about fifty percent of breast cancer diagnosed annually in Taiwan is premenopausal patients, it results in particular consideration to initiate breast cancer screening program in women younger than 50. It is well-­‐documented that mammography screening can decrease the mortality of breast cancer among women older than 50 years old. However, women aged 40 to 49 seem not benefit from mammography screening with the same extent. The Swedish Two-­‐County trial found that the less benefit from screening women younger than 50 compared with those older than 50 is due to a shorter sojourn time and lower sensitivity of mammography in the younger age group. The low sensitivity of mammography could be due to the higher prevalence of dense breasts in young women. Although shortening screening interval may help to detect some cancers earlier, it may not increase mammography sensitivity. The addition of other screening modalities, such as breast ultrasound, may be helpful. One potential false-­‐negative results of ultrasound screening could be due to the difficulty in detecting microcalcification using breast ultrasound. As mammography is the best tool to detect microcalcification-­‐associated ductal carcinoma in situ (DCIS), which probably will not progress into invasive cancer rapidly, it may be feasible to perform mammography screening over a longer interval, with ultrasound added to detect non-­‐palpable cancers not associated with microcalcifications. We conducted a multi-­‐center randomized trial in Taiwan to investigate whether breast ultrasound together with mammography could be an ideal combination for breast cancer screening among women aged 40-­‐49 in Taiwan. Since late 2003, a total of 79,691 female residents aged 40-­‐49 years were invited from community in Taiwan. These participants were first randomly assigned to mammography (n=20040), ultrasound (n=20088), and control group (n=39563). The two former groups were further done by a cross-­‐over design with mammography and ultrasound on alternate year until 2008. The attendance rate of the first round was 59% (11921/20040) for mammography and 56% (11249/20088) for ultrasound. The repeated attendance rate of both groups was 85% in the second round and 91% in the third round. In the first round of screening, the detection rate of breast cancer for the mammography group (0.34%) was 1.5-­‐fold compared with the ultrasound group (0.22%). The additional detection rate was 0.16% contributed from a subsequent ultrasound screening and 0.36% contributed from a subsequent mammogram screening. The combination of mammography with ultrasound was as three to four times as likely to detect breast cancer compared with the control group (annual incidence rate was 0.17%). Our results not only demonstrated higher detection rate and better performance using mammography but also indicated the complementary role of ultrasound applied to breast cancer screening for young Taiwanese women. This further suggests the optimal screening modality for young women in Asian country is to combine mammography with ultrasound. As hand-­‐held breast ultrasound screening is operator-­‐dependent and a real-­‐time examination, standardization and quality control of hand-­‐held ultrasound screening is difficult. Automated whole breast ultrasound (ABUS) may be the key to make ultrasound screening feasible. ABUS has been developed for breast screening in recent years. ABUS could separate image acquisition from image review. In addition, the scanning procedure can be standardized. Therefore, ABUS has the advantage of being operator-­‐independent and is feasible for breast ultrasound screening. So far there are, commercial supine and prone ABUS systems including U-­‐systems, Siemens, Aloka, and iVu ABUSs. The two NTU ABUS systems, GPS ABUS and Robot ABUS, have also been developed by the NTU CSIE and EE Laboratories recently. The GPS ABUS is an US image recoding system developed by the Medical Image Processing Lab of National Taiwan University (NTU). With the sensor response in the magnetic field, the probe position can be tracked and recorded in the system. During the scanning, the probe position relative to the nipple is specified with the degree, clock, and distance. The tracking images and routes are also shown on the screen for real-­‐time reference and are stored for retrospective examination. As a physician still need a lot of time to review the large ABUS images, misdetection might occur. In order to reduce the review time and misdetection by the physician, the computer-­‐aided detection (CADe) for ABUS images has been developed. A region-­‐based ABUS tumor detection method is adopted to provide fast tumor detection. The sensitivities of CADe system could achieve 100%, 90%, and 80%, with the 9.44, 5.42, and 3.33 false-­‐positive rate per view for 134 biopsy-­‐proven lesions, respectively. Talk title: New therapy development for primary breast cancer Speaker: Masakazu TOI Since breast cancers are consisted of heterogeneous subpopulations, the therapeutic strategy should be based on the biological characteristics such as intrinsic subtype. It is also evident that the therapeutic response ranges diversely and unstable. Therefore, breast cancer treatment requires multiple therapeutic modalities, many different types of drugs, often in combination, and response-­‐guided clinical approach. In addition, biomarkers are indispensible for predicting responses and prognosis and for monitoring disease progression and regression. In order to develop new therapeutic agents and therapeutic system, thereby the biomarker-­‐driven and response-­‐guided approach has been investigated extensively especially in neoadjuvant preoperative setting. Pathological responses have been incorporated into major endpoints of clinical trials and several biomarkers are developed for clinical use. Nevertheless, several issues have been also indicated such as that pathological responses do not necessarily predict long-­‐term prognostic outcomes particularly in certain tumor subtypes. It is urgent to consider novel clinical platforms and systems to develop new therapeutic agents and biomarkers.