SPORE in Genitourinary Cancer
University of Texas/ MD Anderson
Colin P.N. Dinney, M.D.
David J. McConkey, Ph.D.
The overall goal of this University of Texas MD Anderson Cancer Center SPORE in Genitourinary Cancer is to facilitate innovative translational research in the prevention, detection, and treatment of this disease leading to the elimination of bladder cancer (BC) as a major health problem. We have invested in several major translational research themes which include: the development of non- or minimally-invasive markers for early detection of BC in high-risk individuals or for surveillance and the early detection of recurrence in those with the disease, the identification of inherited factors that contribute to increased or decreased risk of developing BC, the elucidation of the molecular events (genetic and epigenetic) that mediate the earliest stages of urothelial neoplasia, the determination of whether activating mutations and gene amplifications present in BCs drive cancer progression and serve as potential therapeutic targets, the identification of molecular and biological markers that can be used to distinguish "favorable" from "unfavorable" biology in non-muscle invasive and more advanced disease that direct us to optimal therapy ("personalized medicine"), the development of novel therapeutic approaches for non-muscle invasive BC that are alternatives to bacillus Calmette-Guerin (BCG) and/or are effective in BeG-refractory disease, and the identification of novel agents for the treatment of metastatic BC. To achieve these goals, our SPORE has assembled clinicians and basic scientists including urologists, medical oncologists, pathologists, molecular epidemiologists, molecular and cell biologists, biostatisticians, and experts in development of new technologies and informatics. The SPORE includes 5 inter-related projects that deal with 1) early detection of BC, 2) risk assessment for BC 3) biology and therapeutic targeting of the fibroblast growth factor receptor -3, 4) therapeutic targeting of Ral GTPases, and 5) the development of adenoviral mediated gene therapy for refractory tumors. These projects are supported by 3 Cores: (A) Administrative; (B). Biostatistics and Bioinformatics; and (C) Pathology & Data Management. All of the scientific projects are translational in nature; focus on human BC; involve clinical and basic investigators and biostatisticians; interact with the other projects; and utilize Core resources. Innovative Developmental and Career Development Projects have brought new investigators into and stimulated the SPORE that are represented in each of the major projects. Achievement of the aims and objectives of this proposal will result in a major decrease in the incidence, morbidity and mortality of BC.
PROJECT 1: IMPROVING DIAGNOSIS OF BLADDER CANCER
Bogdan Czerniak, Ph.D.
H. Barton Grossman M.D.
The long-term objective of this project is to improve the detection of one of the common human cancers arising in the bladder by non-invasive voided urine-based tests. Bladder cancer is the 4th most frequent in men and 5th most common overall with approximately 70,500 new cases and 14,600 deaths from the disease in 2009 in the United States. It is estimated that nearly 300,000 patients are regularly monitored in the United States through a variety of non-invasive (urine) and minimally invasive (bladder barbotage, cystoscopy, and biopsy) techniques. Therefore, the development of novel biomarkers that can detect bladder tumors by a non-invasive approach is of major clinical significance. In this grant, we propose to complement the existing Multi-Institutional Aurora kinase A FISH Test Biomarker Validation Trial by novel FISH markers and explore the role of Aurora kinase A in bladder cancer progression. We will compare the specificity and sensitivity of aurora A FISH test complemented by a panel of novel FISH markers with other known noninvasive bladder cancer detection tests for voided urine sediments such as urine cytology and commercially available multi-chromosomal FISH kit known as UroVysion, and NMP22 point of-care test. A panel of novel FISH markers will be developed by whole-organ histologic and genetic mapping (WOHGM) strategy combined with high resolution (1M) Illumina SNP-based genotyping, which provides unique information on the chronology of genomic alterations that parallel the development of bladder cancer from early field effects to invasive disease. Our strong preliminary data indicate that this approach offers a unique opportunity to design novel biomarkers that may address the specific phases of bladder cancer development along these superficial papillary and high grade nonpapillary invasive pathways. Moreover, we propose to investigate the role of aurora kinase A and its regular pathway in bladder cancer progression to invasive disease and distant metastasis. The specific aims of this project are as follows: Specific Aim 1: Develop a panel of novel FISH markers for bladder cancer. Specific Aim 2: Validate a novel set of FISH biomarkers for bladder cancer. Specific Aim 3: Investigate the role of Aurora kinase A in bladder cancer growth, angiogenesis, invasion and metastasis.
PROJECT 2: ROLE OF MICRORNA IN BLADDER CANCER RISK AND OUTCOME: A GENOME-WIDE ANALYSIS
Xifeng Wu, M.D., Ph.D.
Ashish Kamat, M.D. (clinical)
Seth Lerner, M.D. (clinical)
The project builds upon the largest case control study of bladder cancer (BC) in U.S. with extensive epidemiologic and clinical data and rich biospecimens (DNA, tissue, and serum/plasma). We propose a systematic study of microRNAs (miRNAs) in BC etiology, prognosis, and BCG response, including germline SNP genotyping, somatic miRNA profiling, and detection of circulating miRNA. Our specific aims are: 1) To identify novel germline susceptibility loci in miRNA pathway that predispose to BC risk using a two-stage design. We will screen -8000 SNPs in 1000 cases and 1000 controls and then validate top 384 SNPs in an additional 1000 cases and controls. 2) To identify novel germline susceptibility loci in miRNA pathway that predict non-muscle invasive BC (NMIBC) recurrence and progression using a similar two stage design as in Aim 1. In screening phase, we will use 1,200 NMIBC patients from our ongoing case control study. We will also performed stratified analysis on those patients receiving BCG treatment since BCG is the prevalent intravesical therapy for high risk NMIBC. In the validation phase, we will use 300 patients who were enrolled in a clinical trial of BCG treatment. 3) To identify somatic miRNA expression as predictors of BCG response. We will determine global miRNA expression profiles in 50 BC tumor tissues with recurrence and 50 without recurrence in patients receiving BCG treatment to identify somatic miRNA signature that predicts recurrence and then validate the signatures in an additional 75 pairs of tissues. We will also correlate the validated SNPs from Aim 2 with the expression of validated miRNAs from this aim. 4) To identify circulating miRNAs as predictors of recurrence and progression in NMIBC patients receiving BCG treatment. Similar to Aim 3, we will use a two stage design to identify and validate miRNA signatures for recurrence and progression in the context of BCG treatment. Screening will be done in serum of 100 BCG-treated NMIBC patients with and 100 patients without recurrence, and in 50 BCG-treated NMIBC patients with and 50 patients without progression, and validation will be done using 300 patients enrolled in the BCG clinical trial.
PROJECT 3: TARGETING FGFR AND EGFR IN BLADDER CANCER
David J. McConkey, Ph.D.
Colin Dinney, M.D.
Arlene Siefker-Radtke, M.D.
Tumors that express mutant protein kinases are usually dependent upon them for growth and survival. Activating mutations in FGFR3 occur in over half of low-grade non-muscle invasive bladder cancers (BCs) and in a quarter of muscle-invasive tumors, and small molecule and antibody-based FGFR3 inhibitors have exhibited potent growth-inhibitory activities in some BC cell lines and xenografts in preclinical studies. However, clinical translation of these observations has not occurred, in part because dose escalation trials have revealed that FGFR inhibitors produce some toxicity, and whether the extent of target inhibition at non-toxic doses is sufficient to produce apoptosis and/or growth arrest is not clear. We have assembled a collaborative group involving the GU Cancers team at Astra-Zeneca and Dr. Margaret Knowles (University of Leeds, UK) to conclusively determine the value of FGFR3 as a therapeutic target in BC. Our approach will be to use our unique panel of cell lines and xenografts to (1) isolate biomarkers that predict FGFR3 dependency better than FGFR3 mutational status alone and (2) develop pharmacodynamic approaches to determine the extent of tumor FGFR3 pathway inhibition and correlate it with biological response. We will also explore the effects of the novel tumor suppressive "forerunner" gene ARL11 on Ras pathway activation and define the relationships between ARL11 downregulation, FGFR3 and Ras mutational status, and Ras pathway activation in primary tumors, studies that are based on novel findings obtained in Project 1. We will then perform a neoadjuvant clinical trial to determine whether the doses of AZD4547 that can be safely achieved in patients produce sufficient target inhibition to cause apoptosis and/or growth arrest in primary tumors. This methodical approach will provide the strong mechanistic information required for the intelligent design of subsequent Phase II studies in low-grade and muscle-invasive BCs as well as in hematological and other tumors.
PROJECT 4: TARGETING Ral GTPASES IN BLADDER CANCER
Dan Theodorescu, M.D. Ph.D.
David Ross, Ph.D.
Yu Shen, Ph.D.
The last major advance in the treatment of metastatic bladder cancer (BC) took place in 1997 with the advent of gemcitabine. Despite this advance, visceral metastases are usually fatal. The overall goal of the proposed studies is to develop small molecule inhibitors that block a critical node in the metastatic process. We found that Ral GTPases serve as the molecular switches of a therapeutically tractable signaling pathway that allows UC cells to grow in the lung, the most common visceral metastatic site. The clinical significance of this pathway and validity of Ral as a therapeutic target is supported by finding that high Ral expression in tumors places patients at higher risk for metastasis and the requirement of Ral expression for lung metastasis to occur in animal models of UC. Our Guiding Hypothesis for this application is that small molecules targeting RaJ provide effective therapy for metastatic UC. With support from the MD Anderson Bladder SPORE Developmental Research Program (DRP), we evaluated >500K compounds for their ability to bind RaiA or RaiB in computational and combinatorial screens and selected 99 "hits". These were evaluated in a series of secondary assays allowing us to select Ral Binding Compound (RUC)8 and 10 to be pursued in this application. RUC8 and 10 were selected because they: 1) inhibit RaiA to RaiBP1 binding in human UC cells and RaiA induced spreading in murine embryo fibroblasts; 2) inhibit in vitro monolayer growth (IC50 0.5-1.9 1-1M) of human UC cells; 3) bind RaiB directly by nuclear magnetic resonance (NMR) spectroscopy; and 4) have good pharmacokinetic (PK) properties in mice (Cmax 1.3-23 1-1M, T1/2 3.7-4.6 hrs). To develop this novel class of agents we propose the following Specific Aims: Aim 1: Characterize higher potency 2nd generation compounds based on RUC8 and 10 using medicinal chemistry, computational fragment-based design, and similarity search of chemical databases. In the unlikely situation that higher potency compounds are not found in Aim 1, we will pursue Aim 2 and 3 using RUC8 and 10, given their adequate ICSO and in vivo PK. Aim 2: Evaluate 2nd generation compounds for their in vivo therapeutic efficacy in novel human UC models of visceral metastasis. Aim 3: Develop predictive biomarkers of response to antiRal therapeutics in human tissues that will position us for Phase 1 trials by end of this project Documented interest by Astra Zeneca in our work improves overall chances for success in translating our novel Ral inhibitors into the clinical setting as anticancer therapeutics.
PROJECT 5: INTRAVESICAL GENE THERAPY FOR BCG REFRACTORY BLADDER CANCER
William F. Benedict, M.D.
Colin P. N. Dinney, M.D.
Yu Shen, Ph.D.
The overall goal of these studies is to improve the treatment of non-muscle invasive bladder cancer by identifying second line agents that are effective in the scenario of BCG resistance and hopefully for all non-muscle invasive bladder cancer in future years. During the course of the last cycle of SPORE funding, we provided strong evidence for effective gene transfer and potential clinical efficacy when intravesical adenoviral-mediated interferona (IFNa) gene therapy was administered with Syn3 (Ad-IFNa/Syn3). Approximately 50% of the treated patients, all of which were BCG resistant, obtained a complete remission (CR). We also found that Ad-IFNa induces cancer cell death in human bladder cancer cells that are insensitive to the recombinant IFNa protein itself, and that cell death occurs via direct and indirect (bystander) mechanisms. Moreover, normal urothelial cells appear to be resistant to Ad-IFNa. In this renewal we plan to complete a Phase IB study of intravesical Ad-IFNa/Syn3 to optimize the schedule by determining whether therapy on Day 1 and an additional instillation on Day 4 will enhance gene transfer and result in higher and more sustained urine IFNa levels than were achieved with a single treatment. In patients achieving a CR at 90 days we will repeat treatment on Day 1 and 4 to determine whether redosing with this schedule will provide for effective gene transfer. TRAIL, M30 and M65 levels will also be measured in the urine as possible markers of clinical efficacy and tumor cell death. Subsequently we plan to participate in and lead a multicenter Phase II trial based upon the results of Phase IB study.
The isolation and identification of the soluble factor(s) responsible for the Ad-IFNa produced bystander cell death on cancer cells will also be a focus. In addition to its obvious value as a biomarker of Ad-IFN activity, the bystander factor(s) could itself have clinical utility. Various methods will be utilized in these studies, including size exclusion concentration, size-exclusion FPLC column subfractionation and mass spectroscopy.
CORE A: ADMINISTRATIVE CORE
Colin P.N. Dinney, M.D.
David McConkey, Ph.D.
William Benedict, M.D.
The Administrative Core plays a critical organizational role in the success of the SPORE. Dr. Colin Dinney, the Core Director and SPORE Principal Investigator, will facilitate the functioning and interactions of all the projects and the cores. Drs. David McConkey and William Benedict as the Co-Directors of the Administrative Core will co-chair the Executive Committee that is composed of all project Co-Leaders and Core Directors. The SPORE Grant Program Administrator will work closely with Drs. Dinney, McConkey and Benedict to schedule all meetings with investigators and provide timely and effective communication with investigators both within and outside The University of Texas M. D. Anderson Cancer Center. The specific responsibilities of the Administrative Core are: (1) To monitor the research activity of the SPORE; (2) To promote integration and communication between the SPORE and the MD Anderson Multi-disciplinary Research Program in Bladder Cancer (MRP) and the Cancer Center Support (Core) Grant; (3) To monitor scientific integrity and overall compliance with all governmental and NCI regulations and requirements and to coordinate quality assurance including data quality control in conjunction with the Biostatistics and Specimen and Clinical Bio informatics Core; (4) To provide administrative support for the Developmental Research and Career Development Programs; (5) To convene all necessary meetings, including those of the Executive Committee, Internal Scientific Advisory Committee, and External Scientific Advisory Committee; (6) To oversee expenditures and maintain budget information; (7) To communicate and consult frequently with the NCI Project Leader and other staff and prepare all necessary internal and external reports; (9) To establish and maintain electronic communication among all SPORE investigators, including e-mail and a Web page describing SPORE activities, projects, and protocols; (8); To coordinate the activities of the Bladder Cancer Support Team (patient advocacy group); (9) To establish and monitor policies for the recruitment of women and minorities into this program and into any clinical intervention that should arise out of the SPORE; (10) To encourage communication with other groups interested in bladder cancer translational research; (11) To encourage Pharma/Biotech interaction with the SPORE Program.
CORE B: BIOSTATISTICS AND BIOINFORMATICS
Yu Shen, Ph.D.
Keith Baggerly, Ph.D.
The research proposals in this SPORE encompass a broad range of activities, including studies with cell lines, animal models, and clinical trials. These studies will generate different types of data. Properly designing and analyzing such a wide variety of studies will require a variety of statistical and bioinformatics techniques Data and information must flow smoothly between projects. Data quality and integrity must be ensured by using data audit and backup procedures. Also needed is an efficient interface between the computational biology and data storage facilities provided by the SPORE Core C (Pathology and Data Management Core), particularly for the large amounts of microarray and proteomics expression profiling information. To meet these needs, the Biostatistics and Bioinformatics Core brings together several biostatisticians, bioinformaticians and analysts with expertise in a variety of statistical and bioinformatics disciplines. Placing these individuals within the Biostatistics and Bioinformatics Core (rather than in the individual Projects) strengthens the ability of the Projects to interact effectively. This resource also has the flexibility to match personnel to the evolving biostatistics and bioinformatics needs of the SPORE projects. The Biostatistics and Bioinformatics Core will provide expertise in study design and data analysis to all Projects and Cores. The specific aims of the Biostatistics and Bioinformatics Core are to:
The specific aims of the Biostatistics and Bioinformatics Core are to:
Provide guidance in the design and conduct of clinical trials and other experiments arising from the ongoing research of the SPORE.
Provide the innovative and tailored statistical modeling, simulation techniques, and data analyses needed by the Projects, Developmental Research and Career Development Projects, and other Cores to achieve their specific aims.
Ensure that the results of all Projects are based on well-designed experiments and are appropriately interpreted.
CORE C: PATHOLOGY AND DATA MANAGEMENT
Bogdan Czerniak, M.D., Ph.D.
Randal Millikan, M.D.
The task of this Core is to provide well characterized, high-quality human-derived biospecimens required by the projects and to insure that longitudinal, structured, clinical information related to treatment and outcome is collected from the patient donors of those specimens. The primary substrate for these genetic characterization projects is a unique resource consisting of whole-organ maps that will provide histologic characterization and sufficient DNA and RNA to allow multiple experiments to be done on the same samples. This provides an unprecedented opportunity to cross-validate biologic insights coming from these different experimental platforms, and should provide, by far, the most complete view of urothelial carcinogenesis and progression ever realized. This Core will serve as a model for addressing the difficult problems of cross investigator and cross-institutional data exchange, issues of significant interest for the broader context of Texas cancer research. This Core will provide a web-based portal that will allow all investigators to browse and query the contents of the biorepository, de-identified clinical data, and translational research resources such as Standard Operating Procedures, reagent details, and primary datasets. The specific aims for the core are as follows: Specific Aim 1: Provide pathologic review of all tissues. Specific Aim 2: Maintain a urothelial biospecimen repository, Specific Aim 3: Provide professional and technical services for the preparation and quality control of molecular analytes from the biospecimen repository. Specific Aim 4: Abstract clinical information for all patients providing biospecimens. Specific Aim 5: Capture longitudinal clinical follow-up on all patients providing biospecimens. Specific Aim 6: Assure that all information related to the SPORE Project is exchangeable. Specific Aim 7: Provide all SPORE Investigators with a common portal for access to Project data, including support for ad hoc query across projects and domains.