Cryptoprotective Autophagy in Bone Metastatic Prostate Cancer

项目摘要/摘要 应聘者：我目前的职业目标是获得必要的培训和技能，以获得终身教职所需的竞争力，并积累足够的数据，以支持R01或类似的资助机制。我的长期职业目标是成为一名终身教授，一名资助癌症生物学教授，一名导师和一名科学管理人员。为此，我的职业规划的关键要素集中在1)发展我的独立研究，2)继续我的技术和研究管理教育，3)磨练我的沟通能力，4)获得更多的指导经验。 环境：我能够成功地实现我的目标，因为我可以使用莱斯大学、德克萨斯医学中心的最先进的设备、核心设施和教育资源，并通过包括特拉华大学、埃默里大学和锡达斯-西奈大学在内的前列腺癌P01合作项目。我还有一个由杰出导师组成的团队，他们是前列腺癌和癌症健康差距领域的领导者。这些导师包括我的主要导师Mary Farach-Carson博士、共同导师钟丽兰博士、Nora Navone博士和Lovell Jones博士，他们都同意为我提供职业发展指导。 研究：前列腺癌(PCA)是北美男性的第二大死因，那里80%的前列腺癌死亡是由于骨转移。前列腺癌可含有神经元样神经内分泌(NE)的PCA细胞，这与预后不良有关。NE-PCa细胞抵抗凋亡，分泌蛋白质诱导细胞增殖和血管生成，并可去分化促进肿瘤生长。我们实验室最近公布的数据显示，骨髓基质细胞(BMSC)旁分泌信号诱导共培养的骨转移癌PCa细胞凋亡。然而，一组骨转移的PCA细胞可以避免细胞死亡并经历神经内分泌分化(NED)。自噬是一种调节癌细胞存活和细胞分化的过程，也就是细胞内降解和循环的稳态过程。因此，我认为自噬是PCa细胞在骨髓基质微环境中生存和经历NED的一种细胞保护机制。为了解决我的假设，我将(1)剖析暴露在BMSC下的PCa细胞的自噬调节，(2)证明自噬增强PCa细胞的存活并促进NED，(3)证明自噬减弱增加PCa细胞的死亡。自噬的诱导和功能将使用免疫印迹和自噬标记物的染色，如LC3，并使用酸性囊泡染料，如单丹西卡维林进行分析。PCa细胞将受到遗传和药物自噬调节，并检测细胞活力和NED表型。我的研究将确定减弱自噬的途径，并将骨转移的PCA细胞重定向细胞死亡。

Project Summary/Abstract Candidate: My immediate career goals are to obtain the necessary training and skill set required to be competitive for a tenured faculty position and to accumulate data sufficient for an R01 or similar funding mechanism. My long term career goals are to be a tenured, funded cancer biology professor, a mentor, and a science administrator. To this end, the key elements of my career plan are focused on 1) developing my independent research, 2) continuing my technical and research administrative education, 3) honing my communication skills, and 4) gaining more mentoring experience. Environment: I can successfully meet my goals because I have access to state-of-the-art equipment, core facilities, and educational resources at Rice University, the Texas Medical Center, and through a prostate cancer P01 collaboration that includes the University of Delaware, Emory University, and Cedars-Sinai. I also have a team of phenomenal mentors who are leaders in the prostate cancer and cancer health disparities fields. These mentors include my primary mentor, Dr. Mary Farach-Carson, co-mentor, Dr. Leland Chung, and Drs. Nora Navone and Lovell Jones, who have each agreed to provide me with career development guidance. Research: Prostate cancer (PCa) is the second leading cause of death in North American men, where 80% of PCa mortality is due to bone metastasis. PCa tumors can contain neuronal-like neuroendocrine (NE) PCa cells, which correlates with poor prognosis. NE PCa cells are resistant to apoptosis, secrete proteins to induce cell proliferation and angiogenesis, and can de-differentiate to contribute to tumor growth. Our lab recently published data revealing that bone marrow stromal cell (BMSC) paracrine signaling induces apoptosis in co-cultured bone metastatic PCa cells. However, a subpopulation of the bone metastatic PCa cells can avoid cell death and undergo neuroendocrine differentiation (NED). One process that has been shown to mediate cancer cell survival and cell differentiation is autophagy, the homeostatic process of intracellular degradation and recycling. Thus, I propose that autophagy is a cytoprotective mechanism that PCa cells use to survive and undergo NED in the bone marrow stromal microenvironment. To address my hypothesis, I will (1) dissect autophagy regulation in PCa cells exposed to BMSC, (2) demonstrate that autophagy enhances PCa cell survival and promotes NED, and (3) show that autophagy attenuation increases PCa cell death. Autophagy induction and function will be analyzed using immuno-blot and -staining for autophagy markers, such as LC3, and using acidic vesicle dyes, such as monodansylcadaverin. PCa cells will be subjected to genetic and pharmacological autophagy modulation and assayed for cell viability and the NED phenotype. My investigation will identify pathways to attenuate autophagy and redirect bone metastatic PCa cells towards cell death.