Cryptoprotective Autophagy in Bone Metastatic Prostate Cancer

项目总结/摘要 候选人：我的近期职业目标是获得必要的培训和技能，使其具有竞争力，并为R 01或类似的资助机制积累足够的数据。我的长期职业目标是成为一名终身教授，资助癌症生物学教授，导师和科学管理员。为此，我的职业生涯计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）磨练我的沟通技巧，4）获得更多的指导经验。 工作环境：我能够成功地实现我的目标，因为我可以使用赖斯大学、德克萨斯州医学中心的最先进的设备、核心设施和教育资源，并通过包括特拉华州大学、埃默里大学和雪松西奈大学在内的前列腺癌P01合作项目。我还有一个由杰出的导师组成的团队，他们是前列腺癌和癌症健康差异领域的领导者。这些导师包括我的主要导师玛丽·法拉奇-卡森博士、共同导师利兰·钟博士、诺拉·纳沃内博士和洛弗尔·琼斯博士，他们都同意为我提供职业发展指导。 前列腺癌（PCa）是北美男性的第二大死因，其中80%的PCa死亡率是由于骨转移。PCa肿瘤可含有神经元样神经内分泌（NE）PCa细胞，其与不良预后相关。NE PCa细胞对细胞凋亡具有抗性，分泌蛋白质以诱导细胞增殖和血管生成，并且可以去分化以促进肿瘤生长。我们实验室最近发表的数据显示，骨髓基质细胞（BMSC）旁分泌信号诱导共培养的骨转移性前列腺癌细胞凋亡。然而，骨转移性前列腺癌细胞亚群可以避免细胞死亡并进行神经内分泌分化（NED）。已经显示介导癌细胞存活和细胞分化的一个过程是自噬，细胞内降解和再循环的稳态过程。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）剖析暴露于BMSC的PCa细胞中的自噬调节，（2）证明自噬增强PCa细胞存活并促进NED，（3）表明自噬衰减增加PCa细胞死亡。自噬诱导和功能将使用免疫印迹和自噬标记物（如LC 3）染色以及使用酸性囊泡染料（如单丹酰尸胺）进行分析。将对PCa细胞进行遗传和药理学自噬调节，并测定细胞活力和NED表型。我的研究将确定减弱自噬和重定向骨转移性前列腺癌细胞走向细胞死亡的途径。

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.