105年度第二期癌症研究計畫-臺北醫學大學

Core 1 Epidemiological and Bio databank Core Epidemiological and Bio-Bank Core will establish epidemiological and bio-bank core to collect clinical data and tissue samples of breast, colorectal, prostate cancers and brain tumor for the basic research team. Core 2 Cancer Bioinformatics Core There are three cores in our Comprehensive Cancer Center in Taipei Medical University, including (1) Epidemiological and Bio-Bank Core will establish epidemiological and bio-bank core to collect clinical data and tissue samples of breast, colonrectal, prostate cancers and brain tumor for the basic research team; (2)Bioinformatic Core will develop medical informatics integration platform to involved in the four cancer teams; (3) Imaging Core will provide advanced imaging technique to detect tumor site and malignancy and will discover radiogeomic biomarkers for prognosis and treatment effect. Cancer Bioinformatics Core project is concentrated on three major researches. Firstly, to identify the combinations of four type of cancers with common chronic diseases through NHI claim database and to rank the most frequency drugs use with cancers through data mining analysis. Secondary, to evaluate the associations between common chronic diseases and drugs use by computational system biology. Afterwards, to validate and to verify associations of cancer and common chronic diseases/long-term drugs by using cellular/animal model. This research aims to discovery a new therapy for cancer treatment, new service models, and to generate the scientific evidence needed to move the concept of precision medicine into every day clinical practice. Keywords：Common chronic disease, long-term drug use, pharmacoepidemiology, computational system biology, cellular/animal model Core 3 Imaging Core Imaging Core is served as a platform of magnetic resonance imaging (MRI) and computed tomography (CT) to support the four cancer programs in CECR. This imaging research platform can provide the measurements of tumor structure and physiological status in vivo, and therefore facilitate the early diagnosis of tumor, longitudinal monitoring, new drug development, and clinical trial. In next year project, the service and research directions of Imaging Core will focus on three subjects including (1) imaging service and consultation, (2) research team collaboration, and (3) development of advanced imaging and analysis techniques. For the imaging service and consultation to the four cancer programs, the in-vivo imaging of cell, small animal, and human by the 7T MRI at Taipei Medical University, 3T human MRI (service will start from Dec. 2015), and dual-source computed tomography at Taipei Medical University Hospital will be provided. The Imaging Core team, including two radiologists, two assistant professors, two assistant research fellows, and one medical physicist, will assist investigators to resolve the issues of imaging parameters, sequence selection, and image analysis. Regarding the research team collaboration, the collaborations will start from the studies of glioblastoma and extend to all four programs in the perspective of both basic and clinical researches and new drug development. The initial collaborations with four research teams include (a) radiogenomic study of intra- and inter-tumor heterogeneity with Prof. Yung-Hsioa Chiang, (b) imaging of permeability and histologic section to assess the dynamics of blood tumor barrier with Prof. Ruei-Ming Chen, (c) prediction of treatment efficacy of Temozolomide-resistant glioblastoma by MR image phenotypes with Prof. Jian-Ying Chuang, and (d) preclinical animal experiments of a new drug, MPT0B291, for chemical therapy with Prof. Chia-Yi Wang. In the development of advanced imaging and analysis techniques, Imaging Core will keep adjusting and developing the imaging protocols and analysis methods at 7T animal and 3T human MRI, including the Ktrans permeability map for assessing the integrity of blood tumor barrier, diffusion kurtosis imaging (DKI) for elevating the sensitivity in malignant tumor detection, and chemical exchange saturation transfer (CEST) for measuring tumor metabolism. With the achievements of all three subjects, Imaging Core will be able to expedite the progression of cancer researches. Keywords: magnetic resonance imaging, computed tomography, in-vivo imaging, tumor, image biomarkers Program 1 Breast Cancer Research Our previous study demonstrated that smoking-induced breast cancer formation is likely activated through nicotine binding to its receptors. The results implied that the nicotinic receptor should be important for disease formation and therapy. This project is composed of five research themes, which are introduced below. In Theme 1-1, we will continue to establish a large-scale epidemiological data bank for breast cancer patients. We will functionally validate significant single nucleotide polymorphisms (SNPs) related to DNA repair genes. We will also analyze and genotype the SNPs of miR-27a, which was found to be associated with cigarette smoking in our previous result, and its downstream target genes. Additionally, we will explore the gene-gene interaction effect of CHRNA9 and miR-27a and its target genes on the breast cancer risk. In Theme 1-2, an antibody that specifically targets an important protein-C-loop fragment of α9-nAChR has been designed as an antigen by structural analysis. This fragment covers the ligand binding site of α9-nAChR and has induced strong immune responses successfully in experimental chickens. Next, two antibody libraries with high complexity were constructed by phage display technology, and two monoclonal single-chain variable fragments (scFvs), cS7 and cS11, were isolated using panning approaches. The experimental results confirmed the ability of the isolated scFvs to bind to α9-nAChR and to inhibit signal transduction induced by nicotine binding to α9-nAChR. The epitope site of the isolated antibody still needs to be confirmed by epitope mapping, and the complex structure of antibody-α9-nAChR has to be simulated by molecule docking technology. Based on this achievement, theme 1-3 has the following specific aims in 2016: (1) analyze the pharmacokinetics, biodistribution, and in vivo toxicity of BsAb-mPEG nanodrugs (Lipo-Dox® or micelle-quercetin); (2) noninvasively image the α9-nAChR+ tumor targeting efficacy of BsAb-mPEG nanodrugs (Lipo-Dox® or micelle-quercetin); and (3) assess the tumor killing efficacy of BsAb-mPEG nanodrugs (Lipo-Dox® or micelle-quercetin). To evaluate the antitumor efficacy of these nanodrugs, theme 1-4 will established an animal model to explore these possibilities. Theme 1-2 will develop a humanized 9-nAChR-specific antibody for animal experiments. We had a promising result showing that 9-nAChR-specific antibodies S11 and S13 obtain high binding affinity to cancer cells and inhibit cancer cell growth. Furthermore, an in vivo experiment that showed both 9-nAChR antibodies inhibit breast cancer tumor growth and prevent cancer metastasis even in a highly metastatic breast cancer model. Therefore, in 2015, theme 1-4 will take one step forward in investigating whether humanized 9-nAChR antibodies suppress both tumor growth and cancer metastasis. Additionally, a comparison of 9-nAChR knockout breast cancer cells and highly metastatic breast cancer cells will be helpful for determining the anti-cancer mechanism of 9-nAChR antibodies. Theme 1-5 aims to identify the underlying mechanism of α9-nAChR in cancer metastasis and the cancer stemness-related early recurrence of tumors. Our preliminary results from 72 breast cancer tumor tissues showed a significant association between IGF-IR and stemness-related genes such as OCT4, NANOG, CD44, and CD24. Further experiments using a cell model showed that nicotinic treatment is able to induce the expression of α9-nAChR, OCT4, and IGF1R in triple-negative MDA-MB-231 cells; these results were strongly supported by an in vivo metastasis animal model that demonstrated higher expression levels of α9-nAChR, OCT4, and IGF1R in highly metastatic MDA-MB-231 cells. Most importantly, we found that nicotine may stimulate cell migration and stemness-related gene expression through IGF-IR regulation. Therefore, the specific aim of the next year will focus on the effects of specific antibodies and/or small molecules against α9-nAChR or IGF-IR and their associated signaling on nicotine-induced stemness-related properties. Non-metastatic triple-negative MDA-MB-231 and high-migration metastatic MDA-MB-231 cell lines will be used in this study. The expression levels of EMT-related genes and OCT4 will be detected using RT-QPCR and western blot analysis. Furthermore, stemness-related protocols such as drug resistance, side population, 2nd tumor sphere, ALDH activity, and metastasis assays will be performed. The findings of this study will facilitate the development of antibody drugs that could prevent drug resistance and early tumor recurrence by therapeutic targeting of cancer stemness of triple-negative breast cancer. Keywords：Breast cancer, Antibooody, Breast cancer stem cells, Nicotinic acetylcholine receptor, Insulin-growth factor one receptor Program 2 Colorectal Cancer Research Our group has recently reported that about 33.8% of tumor tissues in Taiwan CRC patients contains an APC mutation, which is close to the other reports in Asia, but significantly lower than that in western countries (70-80%). This may indicate the different mechanism of colorectal carcinogenesis existed in Asia and western countries. Our published paper showed that high-mobility group A2 (HMGA2) and miR-21 may potentially serve as biomarkers for predicting aggressive CRC with poor survivability. In pass two years, our preliminary results showed that overexpressed of HMGA2 were associated with miR-21 up-regulation and let-7a down-regulation (aim 2-2-1 and 2-2-3). In addition, CRC patients with high HMGA2 gene expression had poor clinical outcome (aim 2-2-1 and 2-3-1). We also found the levels of miR-194(aim 2-1), Lin28 (aim 2-2-1) and other candidate miRNAs or genes (aim 2-1 and 2-2-2) were associated with HMGA2 expression. In new therapeutic methods, we found that topoisomerase inhibitors, HSp90 inhibitor (NVP-AUY922), were useful to inhibit cell growth of CRC tumor cells with high HMGA2 expression. Next year, we plans to study the candidate CRC miRNA signature, which is regulated by HMGA2, by systems biology approach and also study the regulation of HMGA2/miRNAs interaction in pathogenesis of colorectal cancer patients in different APC gene status. The results will support further understanding of the regulation of cancer biology and the mechanism for CRC carcinogenesis for developing personalized therapy. Keywords：Colorectal cancer, Biomarker, Personalized therapy Program 3 Prostate Cancer Research Prostate cancer research team will evaluate the potential of using AMACR as an early detection tool for prostate cancer diagnosis on high-risk men and compare the performance and cost-effectiveness of AMACR with/or combined with PSA. They will also evaluate the effectiveness of androgen deprivation therapy, develop therapeutic target for CRPC in PC stroma and microenvironment such as Treg and establish a platform for utilizing CTCs as a prognostic marker. Also, they will use the bioinformatics-based query system to build a cohort intergrading clinical, pathological and genetic data promoting future studies in personalized medicine and long-term follow-up. The prostate cancer team will apply the National Health Insurance Database and a longitudinal cohort from Taipei Medical University to analyze the prognostic factors such as pneumonia and ulcerative colitis in prostate cancer. We will apply the AMACR nanobiosensor prospectively in prostate early detection and test it application as a predictor for treatment failure after radical prostatectomy, HIFU and cryoablation. We will conduct a prospective trial to test the hypothesis that local treatment (HIFU) in localized prostate cancer patients (age >75) will prevent the pneumonia incidence in comparison with ADT. CA21 apatmer will be studied in TRAMP and PTEN mice for therapeutic effect in prostate cancer. Characterization of tumor microenvironment specific biomarkers for future lethal phenotypic cancer diagnostic and therapeutic development. Methods: We continue analyze and characterize the molecular mechanisms of L1-CAM, ADAM9-Hemidesmosome and CCR5-CCRL2 interaction in cancer progression and metastasis. We will perform whole-genomic sequence analyses of tumor associated stromal biomarkers for future diagnosis and therapeutic target development. Applying anti-EGFR, anti-Trop2 and anti-Ep-CAM, we have developed a MASC sorting protocol for prostate cancer CTCs. Using our novel 3D co-culture model, we will establish Taipei Prostate Cancer Microtissue Bank. In cooperation with Abnova Co., specific antibodies for AR variants and AR mutants and FISH for AR amplification will be developed to generate a comprehensive analysis of AR expression in CTCs. In addition, we will perform the preclinical evaluation of JM17 in prostate cancer therapy. This platform will be applied in personalized medicine in CRPC treatment. Keywords： Prostate cancer, Tumor microenvironment, Cancer metastasis, Early diagnosis, biomarkers Program 4 Brain Tumor Research Based our results of last year (104) project, we will continue to study new drugs and significant biomarkers for diagnosis, prognosis, and therapy of brain tumors. In drug evaluation, we will focus on the evaluations of nanaoparticle CRLX101 and small molecules MPTOB291 and Sp1 inhibitors for treatment of glioblastoma multiforme (GBM), especially in combined treatment with radiation and TMZ (Thems 4-1 and 4-2). Additionally, we will also evaluate the effects of CRLX101 and MPTOB291 on killing GBM cancer stem cells and TMZ-resistent cells (Them 4-3). In biomarker exploration, a translational radiogenomic study combining the genomic profile and advanced MRI techniques in both human and animal GBM model will be performed to unravel the links between image phenotypes and underlying gene alterations (Theme 4-4); the potentials roles of the circulating miR-106b/miR-25/miR-93 cluster gene as biomarker signatures for diagnosis and prognosis of GBM will be investigated (Theme 4-5); the exosomes from GBM patients will be prepared to examine if blood SAA1 or other proteins can be potentially applied as biomarkers for GBM diagnosis or prognosis (Theme 4-6). Finally, Dr. Tai-Tong Wong, an outstanding pediatric neurosurgeon. In clinical, there is no specific biomarker for prognosis of glioblastomas. He will host a radiogenomic study discovering the relationships between the MR imaging features and genomic profiles in the pediatric glioblastomas malignant brain tumor will be performed. We aim to determine the feasibility of using the pre-operative advanced MR imaging to predict patient outcome and response to treatment by analyzing their radiomic features and genetic changes, and apply to other predict malignant brain tumor in the future(Theme 4-7). This project is expected to clarify the efficacy of anti-brain tumor drugs for clinical trials and clinical treatment, and to find biomarkers for early diagnosis and prognosis. Keywords：Brain tumor, Glioblastoma, Atypical teratoid/rhabdoid tumors, Drug evaluation, Biomarker exploration, Predict malignant brain tumor, Radiogenomics