Validating Conditionally Reprogrammed Cells to Advance Personalized Medicine for Prostate Cancer

Despite decades of research into the causes and possible cures for prostate cancer (PCa), over 26,000 men die each year from this disease in the United States alone. While many of the studies performed over the years have identified important genes and signaling pathways that are involved in prostate carcinogenesis, the fact remains that the shortage of matched patient samples and the limited and suboptimal prostate cell lines available for correlative analyses has hindered the identification of biomarkers that distinguish indolent from aggressive prostate disease and severely impacted the development of effective therapies.Our proposal combines, for the first time, longstanding excellence in urological pathology and clinical PCa treatment with two powerful technologies to directly address the critical unmet needs in PCa research related to the lack of relevant cell lines and the scarcity of matched patient samples.First, we have developed a groundbreaking cell culture methodology that finally allows for the establishment of stable, long-term cultures of both normal and cancerous prostate epithelial cells from extremely small clinical samples. We have termed these cultures conditionally reprogrammed cells, or CRCs, as while the prostate cells have been propagated for well over a year with no gross genetic changes, upon removal from our culture conditions, both the normal and tumor cells cease proliferation, consistent with the primary cells from which they originated. This alone immediately renders our repository the first-of-its-kind inexhaustible biobank of de-identified, annotated, matched (normal and Gleason score 7+) prostate cells.Second, we will analyze the matched CRCs and the primary tissues from which they were derived using a second powerful technology we developed, Reverse Phase Protein Microarrays (RPMA), which generates broad-scale mapping of the activated protein signaling architecture within cells, using extremely small amounts of tissue. This validation and credentialing through extensive proteome profiling is likely to soon lead to advances in identifying markers of aggressive disease. Furthermore, our CRC methodology may lead to profound changes in patient treatment.Specifically, we applied our CRC technology to successfully treat a patient who was dying from a progressive respiratory tumor that required over 300 operations over the past 20 years to keep the patient from asphyxiating. Within 14 days of getting the tissue samples, we identified a drug that was Food and Drug Administration-approved for use in other cancer cases that more effectively at killed his tumor CRCs versus his normal lung CRCs. We also established that the drug he had been scheduled to receive would fail to effectively treat his cancer. Based on our findings, permission was received to place him on the drug we identified. Within 3 months, significant regression of his tumors occurred, which persisted throughout the 12-month treatment schedule and his disease has remained stable for over 7 months after chemotherapy ended (Yuan H, Myers S, Wang J, et al, 2012, The New England Journal of Medicine, 367(13):1220-7). While the application to PCa treatment will require validation of the CRCs, as proposed in our application, we anticipate that our CRC system is a transformational advancement in PCa research.Our proposal is built upon an established and diverse team of collaborators from three top institutions, Georgetown University, Massachusetts General Hospital/Harvard Medical School, and George Mason University's Center for Applied Proteomics and Molecular Medicine. Our team combines longstanding excellence in PCa research, pathology, and clinical PCa treatment and the innovators of high content protein signaling analysis with the inventors of the fundamental breakthrough technology, all of whom have outstanding collaborative and individual PCa research programs. Our team's immediate goal is to fully validate the CRCs in our biobank and to further our research into markers of PCa progression and aggressiveness. We anticipate that the successful completion of our 3-year proposal will not only have an immediate and lasting impact on PCa research via establishment and validation of the biobank and the identification of predicative biomarkers of PCa aggressiveness, but may also lead to profound changes in the way clinical trials are conducted and in the way that new and existing drugs are selected for patient treatment. We expect that our new approach to PCa research eventually may influence the management and treatment virtually all PCa patients.