Prostate Specialized Programs of Research Excellence (SPORE)
Project 1: Role of Gene Fusions in Prostate Cancer
Co-Leader: Arul Chinnaiyan, M.D., Ph.D.
Co-Leader: James E. Montie, M.D.
Employing a bioinformatics approach to analyze prostate cancer gene expression profiles, we identified recurrent gene fusions/translocations in the majority of prostate cancers (Tomlins et al, Science 2005). This represents a landmark discovery emanating from this project and our larger SPORE grant. Specifically, we identified the androgen regulatory elements of TMPRSS2 fused to the members of the ETS family of transcription factors including ERG, ETV1, and ETV4. Analogous to hematological malignancies, gene fusions/translocations identified in prostate cancer may represent pathognomonic biomarkers and molecular sub-types of disease. In this renewal application, we plan to focus our efforts on characterizing this new class of gene fusion biomarkers.
Preliminary work done by our group and others suggest that molecular subtypes as well as transcript variants of gene fusions may be associated with clinical sub-types of prostate cancer. The central hypothesis of this renewal application is that molecular sub-types based on gene fusions and variants will be useful predictors of the aggressive potential of clinically localized prostate cancer and thus guide treatment. Given this, we propose the following Aims:
Specific Aim 1: Characterization of Oncogenic ETS Gene Fusions in Prostate Cancer. The goal of this Aim was to develop the necessary tools to enable the systematic identification of ETS gene fusions in prostate cancers including gene fusion and variants.
Specific Aim 2: Determine the role of ETS family gene fusions in prostate cancer cell lines. Here we proposed to over-express ERG and ETV1 in benign prostate epithelial cells and benign immortalized RWPE cells and monitor their phenotype. Similarly using prostate cancer cell lines, we plan to knock-down ERG gene fusions in VCaP and NCI-H660 cells and ETV1 gene fusions in LNCaP and MDA=PCA2b cells. Various phenotypic readouts will be assessed including cell proliferation, apoptosis, cell invasion, growth in soft agar, and global gene expression profiles.
Specific Aim 3: Characterize the phenotype of androgen-regulated ETS transgenic mice. Here we will systematically analyze the androgen-regulated ERG and ETV1 transgenic mice we created. We will characterize the morphatic lesions harbored in these mice. This assessment will include immnohistochemical analysis as well as global gene expression assessment. We will also cross the ETS transgenic mice with mice in which PTEN is conditionally knocked out in the prostate to determine the cooperative effects of these two important aberrations in prostate cancer progression.
Brenner JC, Ateeq B, Li Y, Yocum AK, Cao Q, Asangani IA, Patel S, Wang X, Liang H, Yu J, Palanisamy N, Siddiqui J, Yan W, Cao X, Mehra R, Sabolch A, Basrur V, Lonigro RJ, Yang J, Tomlins SA, Maher CA, Elenitoba-Johnson KS, Hussain M, Navone NM, Pienta KJ, Varambally S, Feng FY, Chinnaiyan AM. Mechanistic rationale for inhibition of poly(ADP-ribose) polymerase in ETS gene fusion-positive prostate cancer. Cancer Cell. 2011 May 17;19(5):664-78. PubMed PMID: 21575865; PubMed Central PMCID: PMC3113473.
Tomlins SA, Aubin SM, Siddiqui J, Lonigro RJ, Sefton-Miller L, Miick S, Williamsen S, Hodge P, Meinke J, Blase A, Penabella Y, Day JR, Varambally R, Han B, Wood D, Wang L, Sanda MG, Rubin MA, Rhodes DR, Hollenbeck B, Sakamoto K, Silberstein JL, Fradet Y, Amberson JB, Meyers S, Palanisamy N, Rittenhouse H, Wei JT, Groskopf J, Chinnaiyan AM. Urine TMPRSS2:ERG fusion transcript stratifies prostate cancer risk in men with elevated serum PSA. Sci Transl Med. 2011 Aug 3;3(94):94ra72. PubMed PMID: 21813756; PubMed Central PMCID: PMC3245713.
Roychowdhury S, Iyer MK, Robinson DR, Lonigro RJ, Wu YM, Cao X, Kalyana-Sundaram S, Sam L, Balbin OA, Quist MJ, Barrette T, Everett J, Siddiqui J, Kunju LP, Navone N, Araujo JC, Troncoso P, Logothetis CJ, Innis JW, Smith DC, Lao CD, Kim SY, Roberts JS, Gruber SB, Pienta KJ, Talpaz M, Chinnaiyan AM. Personalized oncology through integrative high-throughput sequencing: a pilot study. Sci Transl Med. 2011 Nov 30;3(111):111ra121. doi:10.1126/scitranslmed.3003161. PubMed PMID: 22133722; PubMed Central PMCID: PMC3476478.
Grasso CS, Wu YM, Robinson DR, Cao X, Dhanasekaran SM, Khan AP, Quist MJ, Jing X, Lonigro RJ, Brenner JC, Asangani IA, Ateeq B, Chun SY, Siddiqui J, Sam L, Anstett M, Mehra R, Prensner JR, Palanisamy N, Ryslik GA, Vandin F, Raphael BJ, Kunju LP, Rhodes DR, Pienta KJ, Chinnaiyan AM, Tomlins SA. The mutational landscape of lethal castration-resistant prostate cancer. Nature. 2012 Jul 12;487(7406):239-43. doi: 10.1038/nature11125. PubMed PMID: 22722839; PubMed Central PMCID: PMC3396711.
Prensner JR, Iyer MK, Sahu A, Asangani IA, Cao Q, Patel L, Vergara IA, Davicioni E, Erho N, Ghadessi M, Jenkins RB, Triche TJ, Malik R, Bedenis R, McGregor N, Ma T, Chen W, Han S, Jing X, Cao X, Wang X, Chandler B, Yan W, Siddiqui J, Kunju LP, Dhanasekaran SM, Pienta KJ, Feng FY, Chinnaiyan AM. The long noncoding RNA SChLAP1 promotes aggressive prostate cancer and antagonizes the SWI/SNF complex. Nat Genet. 2013 Nov;45(11):1392-8. doi: 10.1038/ng.2771.Epub 2013 Sep 29. PubMed PMID: 24076601; PubMed Central PMCID: PMC3812362.
Project 2: Preclinical Evaluation and Clinical Development of Potent Small-Molecule Inhibitors of the MDM2-p53 Interaction as a New Therapy for the Treatment of Human Prostate Cancer
Co-Leader: Shaomeng Wang, Ph.D.
Co-Leader: David C. Smith, M.D.
The p53 tumor suppressor plays a central role in controlling cell cycle progression and apoptosis and is an attractive cancer therapeutic target because its tumor suppressor activity can be stimulated to eradicate tumor cells. Recent studies have suggested that stimulation of the p53 activity may be a powerful strategy for the treatment of the majority of hormone-refractory prostate cancer. In p53 wild-type cancer cells, the p53 activity is effectively inhibited by its endogenous inhibitor, the human murine double minute 2 (MDM2) onco-protein by multiple mechanisms. A new therapeutic approach to stimulation of the activity of p53 is through inhibition of its interaction with the MDM2 protein using non-peptide small-molecule MDM2 inhibitors. Design of non-peptide small-molecule inhibitors of the MDM2-p53 interaction is being intensely pursed as a new cancer therapeutic strategy. In the last two years, with the support of the University of Michigan SPORE grant, we have designed and developed a class of highly potent, non-peptide, orally available, small-molecule inhibitors of MDM2. Based upon our promising in vitro and in vivo results, we are advancing our most promising lead compound into human clinical trials as a new therapy for the treatment of human cancer. Our long-term transitional goal in this SPORE renewal project is to develop a highly potent and promising small-molecule inhibitor of the MDM2-p53 interaction (hereafter called MDM2 inhibitor) as a new therapy for the treatment of advanced human prostate cancer. Toward this goal, we will carry out the following specific Aims:
Aim 1: Determination of the in vitro activity, specificity and molecular mechanism of action of our potent MDM2 inhibitors in a panel of prostate cancer cell lines and normal cells.
Aim 2: Determination of the in vivo antitumor activity and molecular mechanism of action of our potent MDM2 inhibitors in animal models of human prostate cancer and examination of any toxicity to animals.
Aim 3: Performance of a Phase II clinical trial of our clinical lead compound in prostate cancer patients with androgen-independent disease. Successfully carried out, this SPORE project will pave the way for the development of an entirely new class of molecularly targeted anti-cancer therapy for the treatment of advanced prostate cancer.
Shangary S, Wang S. Small-molecule inhibitors of the MDM2-p53 protein-protein interaction to reactivate p53 function: a novel approach for cancer therapy. Annu Rev Pharmacol Toxicol. 2009;49:223-41. PMCID: PMC2676449.
Shangary S, Wang S. Targeting the MDM2-p53 interaction for cancer therapy. Clin Cancer Res. 2008 Sep 1;14(17):5318-24. PMCID: PMC2676446.
Yu S, Qin D, Shangary S, Chen J, Wang G, Ding K, McEachern D, Qiu S, Nikolovska-Coleska Z, Miller R, Kang S, Yang D, Wang S. Potent and orally active small-molecule inhibitors of the MDM2-p53 interaction. J Med Chem. 2009 Dec 24;52(24):7970-3. PMCID : PMC2795799.
Zhao Y, Liu L, Sun W, Lu J, McEachern D, Li X, Yu S, Bernard D, Ochsenbein P, Ferey V, Carry JC, Deschamps JR, Sun D, Wang S. Diastereomeric spirooxindoles as highly potent and efficacious MDM2 inhibitors. J Am Chem Soc. 2013 May 15;135(19):7223-34. doi: 10.1021/ja3125417. Epub 2013 May 3. PubMed PMID: 23641733; PMCID: PMC3806051.
Zhao Y, Yu S, Sun W, Liu L, Lu J, McEachern D, Shargary S, Bernard D, Li X, Zhao T, Zou P, Sun D, Wang S. A Potent Small-Molecule Inhibitor of the MDM2-p53 Interaction (MI-888) Achieved Complete and Durable Tumor Regression in Mice. J Med Chem. 2013 Jun 20. [Epub ahead of print] PubMed PMID: 23786219.
Project 3: Defining Genetic Risk Factors for Brothers of Men with Prostate Cancer
Co-Leader: Kathleen A. Cooney, M.D.
Co-Leader: Julie A. Douglas, Ph.D.
While family history is an important risk factor for prostate cancer, localization of highly penetrant prostate cancer susceptibility genes using traditional linkage analysis has been challenging. In this SPORE project, we used our ongoing study of hereditary prostate cancer study (the University of Michigan Prostate Cancer Genetics Project or PCGP) to identify a set of sibling pairs discordant for prostate cancer. These siblings can be used to implicate genes of modest penetrance using family-based association methods. Since the sibships are derived from families with early-onset and/or hereditary prostate cancer, they are relatively enriched for genetic susceptibility factors. During the first five years of funding, we have established the discordant sibling pair (DSP) project as a resource for characterizing germline variants associated with prostate cancer. To date, we have studied 14 candidate genes and have shown that single nucleotide polymorphisms (SNPs) in CYP17, BRCA1, FHIT, SDF1, CXCR4, and AMACR are significantly associated with prostate cancer. Our most compelling association finding involves a glutamine-to-arginine substitution at codon 356 (Gln356Arg) in exon 11 of the BRCA1 gene that accounts for some (but not all) of our prior evidence of prostate cancer linkage to chromosome 17q21 in a PCGP genome-wide linkage scan. Non-synonymous SNPs (nsSNPs), such as BRCA1 Gln356Arg, result in single amino acid substitutions and have been shown to account for many of the genetic changes that influence Mendelian disorders. In this SPORE renewal project, we propose a genome-wide approach focusing on nsSNPs in known genes, including many genes previously implicated in cancer. This proposed genome-wide approach has the advantage of testing for variants that are likely to be causative and has been successfully used to identify novel candidate loci for type 1 diabetes and Crohn disease.
To test the hypothesis that common nsSNPs in BRCA1 and other candidate genes are associated with prostate cancer, the following two Specific Aims are proposed:
Specific Aim 1: Develop our new, formal collaboration with the SPORE program at Johns Hopkins University to follow-up and generalize significant prostate cancer associations, including our previously reported prostate cancer association with BRCA1 Gln356Arg.
Specific Aim 2. Complete a replication-based genome-wide association study of early-onset and familial prostate cancer using more than 11,500 nsSNPs that cover approximately 6,500 known human genes, including a disproportionate number in cancer-related pathways.
Ewing CM, Ray AM, Lange EM, Zuhlke KA, Robbins CM, Tembe WD, Wiley KE, Isaacs SD, Johng D, Wang Y, Bizon C, Yan G, Gielzak M, Partin AW, Shanmugam V, Izatt T, Sinari S, Craig DW, Zheng SL, Walsh PC, Montie JE, Xu J, Carpten JD, Isaacs WB, Cooney KA. Germline mutations in HOXB13 and prostate-cancer risk. N Engl J Med. 366(2):141-9, 2012. PMID:22236224. PMCID: PMC3779870.
Zuhlke KA, Johnson AM, Okoth LA, Stoffel EM, Robbins CM, Tembe WA, Salinas CA,Zheng SL, Xu J, Carpten JD, Lange EM, Isaacs WB, Cooney KA. Identification of a novel NBN truncating mutation in a family with hereditary prostate cancer. Fam Cancer. 2012 Dec;11(4):595-600. doi: 10.1007/s10689-012-9555-1. PubMed PMID: 22864661; PubMed Central PMCID: PMC3485445.
Raymond VM, Mukherjee B, Wang F, Huang SC, Stoffel EM, Kastrinos F, Syngal S,Cooney KA, Gruber SB. Elevated risk of prostate cancer among men with Lynch syndrome. J Clin Oncol. 2013 May 10;31(14):1713-8. doi: 10.1200/JCO.2012.44.1238.Epub 2013 Mar 25. PubMed PMID: 23530095; PMCID: PMC3641694.
Xu J, Lange EM, Lu L, Zheng SL, Wang Z, Thibodeau SN, Cannon-Albright LA, Teerlink CC, Camp NJ, Johnson AM, Zuhlke KA, Stanford JL, Ostrander EA, Wiley KE,Isaacs SD, Walsh PC, Maier C, Luedeke M, Vogel W, Schleutker J, Wahlfors T,Tammela T, Schaid D, McDonnell SK, DeRycke MS, Cancel-Tassin G, Cussenot O, Wiklund F, Gr�nberg H, Eeles R, Easton D, Kote-Jarai Z, Whittemore AS, Hsieh CL,Giles GG, Hopper JL, Severi G, Catalona WJ, Mandal D, Ledet E, Foulkes WD, HamelN, Mahle L, Moller P, Powell I, Bailey-Wilson JE, Carpten JD, Seminara D, Cooney KA, Isaacs WB; International Consortium for Prostate Cancer Genetics. HOXB13 is a susceptibility gene for prostate cancer: results from the International Consortium for Prostate Cancer Genetics (ICPCG). Hum Genet. 2013 Jan;132(1):5-14.doi: 10.1007/s00439-012-1229-4. Epub 2012 Oct 12. PubMed PMID: 23064873; PMCID: PMC3535370.
Schroeck FR, Zuhlke KA, Siddiqui J, Siddiqui R, Cooney KA, Wei JT. Testing for the recurrent HOXB13 G84E germline mutation in men with clinical indications for prostate biopsy. J Urol. 2013 Mar;189(3):849-53. doi: 10.1016/j.juro.2012.09.117. Epub 2012 Oct 2. PubMed PMID: 23036981.
Project 4: Inhibition of CCL2 to Treat Metastatic Prostate Cancer
Co-Leader: Kenneth J. Pienta, M.D.
Co-Leader: Maha Hussain, M.D.
We have identified monocyte chemoattractact protein-1 (MCP-1, CCL2) as a novel potent regulator of prostate cancer proliferation and migration. The ability of CCL2 to influence prostate cancer (PCa) tumorigenesis and metastasis may occur via direct promotional effect on tumor cell growth and function as well as a modulatory effect on the tumor microenvironment by promoting macrophage mobilization and infiltration into the tumor bed. We have demonstrated that PCa cells in vitro and in human cancer tissues exhibit an upregulation of the CCL2 receptor, CCR2. In addition, a major role of CCL2 on tumor growth and metastasis has been linked to its regulatory role in mediating monocyte / macrophage infiltration into the tumor microenvironment and stimulating a phenotypic change within these immune cells to promote tumor growth (tumor associated macrophages, TAMs). CCL2 has previously been shown to be an important determinant of monocyte / macrophage infiltration in breast, cervix and pancreatic carcinomas and the levels of CCL2 expression have been correlated with the involvement of lymphocyte and macrophage localization in secondary sites of tumor formation. We were the first to establish a direct stimulatory role of CCL2 on PCa cells in vitro. Utilizing anti-human (CNTO888) and anti-mouse (C1142) specific neutralizing antibodies to CCL2, we have demonstrated an inhibition of prostate tumor growth and migration in vivo through direct effects on the PCa cells as well as blocking the infiltration of TAMs into the tumors. The availability of the human antibody CNTO888 to CCL2 will allow us to test our observations and hypothesis in humans: Overall Proposal Hypothesis: Systemic inhibition of monocyte chemoattractant protein -1 (MCP-1; CCL2) will be an effective treatment for prostate cancer. To test this hypothesis we will perform the following specific aims: 1) We will dissect the role of increased CCL2 expression on monocyte mobilization in response to prostate cancer, 2) we will dissect the role of CCL2 on macrophage infiltration and subsequent tumor growth and metastasis of prostate cancer, and 3) we will test the human antibody CNTO888 to CCL2 in patients with prostate cancer. Completion of these experiments will define the role of infiltrating macrophages in prostate cancer biology and characterize the validity of targeting CCL2 for the treatment of advanced prostate cancer.
Zhang J, Patel L, Pienta KJ. Targeting chemokine (C-C motif) ligand 2 (CCL2) as an example of translation of cancer molecular biology to the clinic. Prog Mol Biol Transl Sci. 2010;95:31-53. doi: 10.1016/B978-0-12-385071-3.00003-4. Review. PubMed PMID: 21075328; PubMed Central PMCID: PMC3197817.
Zhang J, Lu Y, Pienta KJ. Multiple roles of chemokine (C-C motif) ligand 2 in promoting prostate cancer growth. J Natl Cancer Inst. 2010 Apr 21;102(8):522-8. doi: 10.1093/jnci/djq044. Epub 2010 Mar 16. Review. PubMed PMID: 20233997; PubMed Central PMCID: PMC2857800.
Roca H, Varsos ZS, Pienta KJ. CCL2 is a negative regulator of AMP-activated protein kinase to sustain mTOR complex-1 activation, survivin expression, and cell survival in human prostate cancer PC3 cells. Neoplasia. 2009 Dec;11(12):1309-17. PubMed PMID: 20019839; PubMed Central PMCID: PMC2794512.
Zhang J, Patel L, Pienta KJ. CC chemokine ligand 2 (CCL2) promotes prostate cancer tumorigenesis and metastasis. Cytokine Growth Factor Rev. 2010 Feb;21(1):41-8. doi: 10.1016/j.cytogfr.2009.11.009. Epub 2009 Dec 14. Review. PubMed PMID: 20005149; PubMed Central PMCID: PMC2857769.
Mizutani K, Sud S, McGregor NA, Martinovski G, Rice BT, Craig MJ, Varsos ZS, Roca H, Pienta KJ. The chemokine CCL2 increases prostate tumor growth and bone metastasis through macrophage and osteoclast recruitment. Neoplasia. 2009 Nov;11(11):1235-42. PubMed PMID: 19881959; PubMed Central PMCID: PMC2767225.
Roca H, Varsos ZS, Sud S, Craig MJ, Ying C, Pienta KJ. CCL2 and interleukin-6 promote survival of human CD11b+ peripheral blood mononuclear cells and induce M2-type macrophage polarization. J Biol Chem. 2009 Dec 4;284(49):34342-54. doi:10.1074/jbc.M109.042671. Epub 2009 Oct 15. PubMed PMID: 19833726; PubMed Central PMCID: PMC2797202.