PROJECT SUMMARY The prevention of invasion and metastasis remains paramount for the reduction of prostate cancer mortality. Locally confined tumors are rarely lethal and survival is all but assured until metastasis occur, thus understanding the mechanisms driving invasion are critical. Once possible driver is hypoxia, or low oxygenation. We do not know the biological mechanisms by which hypoxia drives metastasis and currently have no effective therapeutic options to prevent it. However, in this research proposal, we have reason to believe that hypoxia-induced PIM is leading to increased invasion and metastasis through regulation of Abi2 and actin polymerization. To test this hypothesis, we will determine how hypoxia modulates Abi2 levels and WRC formation. To do this we will investigate the effects of hypoxia-induced PIM phosphorylation on Abi2, determine effects of hypoxia and PIM on the WRC, and characterize the effect of hypoxia on actin dynamics. Additionally, we will investigate the role of PIM in hypoxia-induced invasion. The goal is to determine whether hypoxia-induced PIM is sufficient to drive invasion in vitro and understand the role of PIM in determining invasive potential of cells in vivo. These data should help in establishing a novel connection between PIM kinases and regulation of cell motility via cytoskeletal rearrangement in hypoxia. Elucidating mechanisms regulating motility and invasion are critical for understanding metastatic disease progression. In doing this project, we will establish a novel prostate-specific invasion model to better recapitulate what occurs in human disease. This will allow for better clarity into the metastatic process and ultimately should provide insight into how to improve overall patient survival. These results will elucidate the potential therapeutic role for PIM inhibitors and how they could be used to increase the efficacy of patient treatment. Lastly these data will also yield insight into mechanisms of how hypoxia increases invasion that could be exploited in the clinic and make a direct impact on patients care. In summary, the research proposed in the grant will elucidate mechanisms by which hypoxia is a driving factor in metastasis and provide insight into pathways that serve as viable drug targets to improve efficacy of patient treatment while simultaneously increasing overall patient survival.