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Adult Oncology, Dana-Farber Cancer Institute
Project 1 Loss of PTEN expression is associated with increased biologic potential of aggressive behavior in prostate cancer. Both the IGF-1 axis and PTEN act on the PI3K/Akt signaling pathway, and activation of this pathway enhances prostate carcinogenesis through a variety of mechanisms, including increasing proliferation. One target of this signaling pathway is the cyclin- dependent kinase inhibitor p27. Loss of p27 expression is associated with poor prognosis in prostate cancer. Our goal is to extend our current understanding of prostate cancer to specific molecular alterations that determine biologic potential. Thus, to complement the extensive clinical, serological and nutritional database we already assembled within the Physicians' Health Study, we propose to obtain and archive paraffin-embedded prostate cancer tissue blocks from 828 cases of prostate cancer from 1982 to 2002. In this project, we will assess standard histopathology, PTEN and p27 expression, and proliferation. We will examine whether the prevalence of loss of PTEN and p27 expression in prostate cancers is decreased in cancers detected through PSA screening, and we will compare the prevalence of loss of PTEN and p27 in tumors from African-American men and Caucasian men. We will also examine whether loss of PTEN expression is related to loss of p27 expression, increased proliferation, and worse prognosis (mortality, and PSA relapse) in prostate cancer, and whether loss of p27 expression is related to increased proliferation, and worse prognosis. Using our serological samples, we will examine whether high level of plasma IGF-1 and low level of IGFBP-3 are related to decreased p27 expression, and increased proliferation in prostate cancer. Finally, we will examine whether markers of prostate cancer progression (loss of p27 expression, and increased proliferation) are increased, and prognosis worsened, by high level of plasma IGF-1 and low level of IGFBP-3 primarily in tumors without loss of PTEN expression. The tumor markers we plan to assess in this project, as well as future novel markers discovered through other SPORE projects, will provide the basis for future studies of prognostic indicators. Specific Aims
Project 2 The nuclear receptor PPAR_ is a dominant regulator of adipose differentiation and a modulator of the growth of many cell types. It is activated by synthetic ligands including the synthetic thiazolidinedione (TZD) drugs, such as rosiglitazone. Recent data, from our lab and others, indicates that PPAR_ activation can inhibit the growth of epithelial cells from prostate, breast and colon, and change patterns of gene expression toward a more differentiated phenotype. Small clinical trials in human prostate cancer have shown that rosiglitazone can cause a prolonged stabilization in PSA levels in a subset of human patients. This has led to an additional, larger clinical trial; this proposal is intended to help support and advance the planning and interpretation of human clinical trials for the use of PPAR_ ligands in CaP. Our first Aim will perform transcriptional profiling in human CaP cells treated with PPAR_ ligands. We will pay particular attention to genes, which encode cell surface or secreted proteins, as these could serve to measure PPAR_ activation. Our second Aim will study the genetic status of the PPAR_ gene in the patients in the DF/HCC clinical trials. In particular, it will be important to correlate responsiveness in the patients with expression levels and potential mutations or deletions in PPAR_. Our final Aim will perform experiments in mice that will model the treatment of CaP with PPAR_ ligands. We will first examine the effects of PPAR_ mutations on the propensity toward CaP in mice with mutations in PTEN and P21. These mice will then be used to examine the effects of a PPAR_ ligand (rosiglitazone) before and during the development of cancer. These studies together will provide useful knowledge that may eventually lead to new methods to prevent or treat human CaP. Specific Aims
Project 3 Our group has approached the problem of prostate cancer classification and stratification using genome-wide expression analysis. Our recent data shows that expression patterns derived from a study of 12,000 human cDNA¹s in 52 prostate tumors and 50 normal ³non-tumor² prostate samples can reliably distinguish tumor from normal, low from high Gleason Score tumors, and can accurately predict relapse following prostatectomy. We now wish to discover novel gene expression patterns that are linked to the underlying genetic abnormalities found in these tumors. It is our hypothesis that such gene expression patterns will allow one to discover those genes and gene products that are casually linked to tumor development and progression. To this end will use high-density Single Nucleotide Polymorphisms (SNP) arrays to analyze genetic loss in tumors for which expression data is already in-hand. In aims 1 and 2 we will use high-density SNP arrays to determine genome-wide patterns of genetic loss in 52 prostate tumors for which gene expression data is already known. Using supervised methods of data analysis we will try to discern gene expression patterns that are correlated with regional loss-of-heterozygosity. This may allow the identification of the pathways casually linked to prostate cancer development, and ultimately to the identification of novel cancer therapeutic targets. We are able to use high-density array based methods of SNP analysis to discern the patterns of genetic loss in tumors for which only paraffin embedded tissue sections are available. This technology can thus be used to describe and categorize cancers wherein only fixed biopsy specimens exist. Therefore, in aim 3 we will use this technology to test the hypothesis that differences in the genetic composition of tumors in patients undergoing radiotherapy account for differences in patient outcome. Specific Aims
Project 4 The overall goal of Project 4 is the development of a gene expression-based predictor of outcome in prostate cancer. The hypothesis underlying this project is that the clinical heterogeneity of prostate cancer has a molecular signature, but that signature has yet to be identified. We will focus on the heterogeneity of outcome following radical prostatectomy of patients with organ-confined disease, and we will use DNA microarrays to identify the gene expression signature of propensity for tumor recurrence. In Aim I, we will generate a high quality, clinically and pathologically annotated gene expression database of 150 primary tumors for which long-term clinical follow-up is available. RNAs generated from these frozen tumors will be subjected to DNA microarrays containing probes for 60,000 genes and ESTs. In Aim II, we will use this gene expression database, together with clinical information, to generate a molecular predictor of recurrence. We will use supervised machine learning approaches to this problem, evaluating the accuracy of the outcome predictor by several criteria including random permutation testing, leave-one-out cross-validation testing, and testing on an independent dataset. In Aim III of the proposal, we will work to develop a version of the outcome predictor that can be implemented in a more routine clinical setting, rather than in a highly specialized genomics research laboratory. To that end, we will explore extending the outcome prediction model to an immunohistochemical implementation, to a quantitative RT-PCR approach, and we will work to develop methods suitable for extracting this information from small, needle biopsy prostate specimens. At the time of completion of this project we aim to have developed a robust molecular predictor of outcome in prostate cancer, and to have translated this assay into one which can be evaluated by the wider community. It is anticipated that such molecular predictors of clinical behavior will be of value in individualizing treatment decision-making for patients with prostate cancer. Specific Aims
Project 5 The majority of prostate cancer (PCa) are androgen dependent and respond to androgen ablation therapy, but most relapse with disease that is clinically androgen independent. However, the androgen receptor (AR) is still expressed in androgen independent prostate cancer (PCa) and there appears to be selective pressure for AR gene amplification and mutations that augment AR activity. These observations suggest that the AR remains a therapeutic target in androgen independent PCa and that our current hormonal therapies may be inadequate to suppress AR function. To test this hypothesis we have developed androgen independent PCa cell lines from the CWR22 xenograft that are also AR antagonist (bicalutamide) resistant, but express AR and prostate specific antigen, consistent with the phenotype of clinical androgen independent PCa. This project will use these and additional xenografts to assess the role of the AR in androgen independent PCa and to develop new therapies that target the AR in androgen independent PCa. Xenografts and clinical samples will then be examined by oligonucleotide arrays to determine mechanisms of androgen independent growth and indentify groups of genes whose expression predicts responses to AR directed therapies. The specific aims are to 1) determine whether the AR is active and necessary for tumor growth or survival in androgen independent PCa cell lines, 2) determine whether the AR contributes to the in vivo growth of a series of androgen independent PCa xenografts, and 3) subset androgen independent PCa xenografts based upon responses to therapy and compare with clinical samples using oligonucleotides microarrays. Taken together, the results from these experiments will establish whether the AR is active and a therapeutic target in androgen independent PCa, identify mechanisms of androgen independent growth, and identify subsets of patients likely to respond to further AR directed therapy. Specific Aims
Core 1 The purpose of the Administration, Evaluation and Planning Core is to assure the coordination of the Dana Farber/Harvard Cancer Center (DF/HCC) Prostate Cancer SPORE components and to provide oversight and leadership of the scientific, administrative and fiscal aspects of the SPORE. Within the DF/HCC Prostate Cancer SPORE, there are several layers of oversight and evaluation. Dr. Kantoff, as SPORE Director, will monitor the progress of the Projects and Cores, and oversee the Career Development and Developmental Projects Programs and oversee all other proposed activities. Our Governance Committee, made up of senior members of the DF/HCC Prostate Cancer Program, will meet monthly to provide immediate decision-making. The Internal Advisory Board, comprised of prominent members of the Harvard Medical School community, and representing the participating institutions and major cancer research disciplines. Our External Advisory Board will meet in the Boston area annually during the five-year funding cycle. The responsibilities of this core are to: 1) Monitor research progress and plan for the future; 2) Foster collaborative research within the SPORE and between SPOREs; 3) Integrate the Prostate SPORE into the DF/HCC structure; 4) Provide necessary resources and fiscal oversight; 5) Promote rapid dissemination of significant research findings. SPECIFIC AIMS The Specific Aims of the Administration, Evaluation and Planning Core are to:
Core 2 The Biostatistics Core supports all research activities within the SPORE, including Projects, Developmental Projects and other Cores. The Core will provide collaboration and consultation on study design, data management and quality control, and data analysis and interpretation to SPORE researchers. The specific aims are to:
Core 3 The Tissue and Pathology Core has two specific aims. The first specific aim is to provide a tissue and blood repository for use by SPORE investigators. Two independent tissue banks have been previously established at three of the performance institutions in this SPORE. A joint Brigham and Women's/Dana-Farber Cancer Institute Bank (BWH/DFCI Prostate Tissue Bank) and a bank at the Beth Israel Deaconess Medical Center. These tissue resources together with the DFCI Blood Component Repository will together comprise the SPORE Tissue and Blood Repositories. The tissue and Pathology Core will collect, freeze, and store a) fresh samples of prostate cancer and paired non-tumor prostate tissue; b) blood and blood components (plasma, peripheral blood mononuclear cells from patients with prostate cancer). The Tissue and Pathology Core will also identify, collect and provide investigators with samples of fixed tissues from biopsy and prostatectomy specimens of patients who are consented, to allow analysis of these tissues in combination with associated clinical data. The second major specific aim of this Core is to provide research pathology services to SPORE investigators, including histology, immunohistochemistry, in situ hybridization, computer-assisted image analysis, laser capture microdissection, and the generation of and access to tissue microarrays. The Tissue and Pathology Core will create a seamless informatics link between the extensive existing tissue resources among the Dana Farber/Harvard Cancer Center (DF/HCC) hospitals. Thus, one of the major goals of this core facility is to create a tissue and blood resource, linked to clinical outcome data, behind a secure data management system that will be available to DF/HCC SPORE investigators as well as SPORE investigators at other institutions. In this regard there will be close collaboration between the Tissue and Pathology Core and the Biostatistics Core. The Biostatistics core will be responsible for assisting in the data analysis and data auditing. Specific Aims
Core 4 The overall purpose of Core 4 (Genomics Core) is to perform microarray-based genomics experiments for the SPORE Projects. Specifically, DNA microarray-based gene expression profiling will be performed for the projects, including assisting Project leaders with experimental design, preparing labeled RNAs, performing microarray hybridizations, and assisting the Projects with bioinformatics analysis of the data. Similarly, we will perform microarray-based single nucleotide polymorphism (SNP) experiments for Projects requiring whole genome SNP LOH analyses. The Core will thus perform these genomic functions for which specific technical expertise is required, uniformly high data quality is essential, and for which economies of scale clearly exist. Specific Aims
Career Development Project The Prostate Cancer Program of the Dana Farber/Harvard Cancer Center seeks applications for a Career Development Award in Prostate Cancer Research. Funding has been provided through a grant from the National Cancer Institute in the form of a Specialized Program of Research Excellence (SPORE), awarded to the Prostate Cancer Program of DF/HCC. Eligible candidates are those who are in their final year of clinical fellowship or postdoctoral fellowship (MD, PhD or MD/PhD required). A track record of interest and productivity in prostate cancer research is required. The awardee will receive up to two years of funding at $50,000 per year, which can be used for salary and/or support of research at Harvard Medical School, the Harvard School of Public Health or any of the Harvard-affiliated hospitals. The individual would become a member of the DF/HCC Prostate Cancer Program and the DF/HCC Prostate Cancer SPORE. Women and minority candidates are encouraged to apply. Please forward requests for applications to Dr. Philip Kantoff, Dana-Farber Cancer Institute, 44 Binney St., Boston, MA 02115 by September 16, 2002. Inquiries should be addressed to Kathleen Keavany, SPORE Administrator at 617-632-3866.
Developmental Projects Program The Prostate Cancer Program of the Dana Farber/Harvard Cancer Center seeks applications for funding of research on prostate cancer. Funding will be provided through the Specialized Program on Research Excellence (SPORE) mechanism, a National Cancer Institute program. Applicants must be on the faculty at Harvard Medical School at the level of Instructor or higher. While the proposed research can be basic or clinical, bench to bedside and bedside to bench strategies need to be addressed, since the purpose of this program is to promote translational research in prostate cancer. Applications will be judged on their scientific merit and/or the potential for contribution to the overall status of the DF/HCC Prostate Cancer Program/Prostate Cancer SPORE. Funding will be provided on a renewable one-year basis. Funding will be for up to $50,000 in total costs per year. Applications should not exceed five pages, not including references, and should be submitted by September 16, 2002 to Dr. Philip Kantoff, Dana Farber Cancer Institute, 44 Binney Street, D1230, Boston, MA 02115. A prior track record in prostate cancer research is preferred but not necessary. Minority and women applicants are encouraged to apply. Inquiries should be addressed to Kathleen Keavany, SPORE Administrator at 617-632-3866. |
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