Capacitation involves biochemical and physiological processes in sperm with motility changes (including hyperactivation) that allow sperm to bind to the zona pellucida, undergo acrosome reaction and penetrate the ovum. Bovine serum albumin (BSA), has been thought to facilitate capacitation in mouse sperm through depleting the sperm membrane of cholesterol. Other albumin preparations including human serum albumin (HSA), fatty acid free (FAFHSA), and recombinant albumin (RHSA) may affect capacitation differently. Moreover, cholesteryl ester transfer protein (CeTP) may facilitate capacitation but it should not be present in yeast-derived RHSA as opposed to human blood-derived HSA; RHSA and HSA may support capacitation at different extents. These preparations were tested in motility experiments after 3-5.75 hr (mean 4 hr) capacitating incubation. Sperm were purified by 40/80% PureSperm gradients and washed in HTF-BSA, then diluted in final albumin to 10-20M/mL. FAFHSA, BSA, HSA, and RHSA had hyperactivation percentages of 2.83%, 3%, 11.3%, and 15.75% respectively. RHSA had a motility of 71.3% on average while the others ranged from 83.6% to 88%, affecting the hyperactivation results. RHSA also caused incubated sperm to agglutinate or aggregate, but its elevated hyperactivation rates relative to HSA are insignificant. These results fit previous data suggesting that BSA and FAFHSA do not support hyperactivation (therefore, capacitation) as both are abnormally below 10% hyperactivation. Since BSA contains low CeTP, this suggests CeTP may help facilitate capacitation. Previous studies demonstrated the amount of CeTP in albumin correlated well with albumin’s ability to support acrosome reaction. The present results suggest that capacitating motility patterns may not necessarily correlate with the ability to acrosome react; this hypothesis could be tested. Since fertilization is largely misunderstood; understanding the roles of Albumin or CeTP may aid in developing infertility treatments, contraceptives and family planning for a growing human populace.

Early diagnosis is one of the key factors to decrease disease-related mortality of advanced prostate cancer. Prostate-specific antigen (PSA) has been utilized as a diagnostic marker for prostate cancer. However, PSA test can cause false positive because enlarged or inflamed prostates also cause increased PSA level. Therefore, new markers are needed to improve the diagnostic accuracy. To discovery novel markers for advanced prostate cancer, we analyzed prostate fluids by mass spectrometry proteomic analysis. We discovered that the regenerating islet-derived protein 3 gamma (Reg3g) is specifically and highly expressed in the prostatic fluids of prostate cancer bearing mice. Reg3g has been shown upregulated in pancreatic cancer and can promote pancreatic cancer progression via STAT3 pathways. Additionally, overexpression of Reg3g induced immunosuppressive tumor microenvironment by recruiting myeloid-derived suppressor cells and enhancing CTLA-4 and PD-1 levels in T cells. Based on these observations, we hypothesize that Reg3g is a novel diagnostic marker and therapeutic target for advanced prostate cancer. We seek to 1) Determine whether overexpression of Reg3g affects prostate tumor cell proliferation and metastasis in vitro. 2) generate a prostate specific Reg3g overexpression transgenic mouse model to investigate whether Reg3g promotes prostate cancer progression and how it affects immune cells infiltration into tumor. Therefore, if we demonstrate that Reg3g expression is closely correlated prostate cancer progression and metastasis, this study will have an immediate impact on diagnosis and treatment of prostate cancer. This proposal addresses two overarching challenges: 1) Develop treatments that improve outcomes for men with lethal prostate cancer and 2) Define the biology of lethal prostate cancer to reduce death.