Vascular Complications of Polycystic Kidney Disease

描述(由申请人提供)： 钱博士的长期目标是帮助更好地了解肾脏疾病的病理生理学，特别是常染色体显性遗传性多囊肾病(ADPKD)的血管并发症。她的目标是通过实施这项提议，在五年内成为一名训练有素的独立内科科学家，在遗传学以及分子和细胞生物学方面拥有先进的理论和技术知识。候选人的导师Torres博士是ADPKD临床和血管方面的专家，她的共同导师Harris博士带来了ADPKD遗传学的详细知识。第三位导师法鲁贾博士是钙离子动态平衡方面的专家，他将为提案中的这一部分提供有价值的指导。这项建议的重点是研究ADPKD蛋白、多囊蛋白-1和-2在血管系统中的作用，并确定这些蛋白的缺陷是否与ADPKD相关的血管表现和高血压直接相关。初步研究表明，多囊蛋白在血管平滑肌细胞(VSMC)中表达，它们作为多囊蛋白复合体的一部分相互作用，可能起到维持细胞内钙离子的作用。本方案的前两个特定目标是研究小鼠Pkd1和PKD2基因突变以及人类蛋白在小鼠体内过度表达对VSMC特性的影响。这些将通过研究突变小鼠血管系统的超微结构，在体外进行分析，作为培养的VSMCs和在体内进行分析。具体目标3将使用成像方法、免疫共沉淀和酵母双杂交技术鉴定VSMC多囊蛋白复合体的成分。这些研究的结果将使VSMC多囊蛋白复合体得以定义，并与上皮细胞中的相应复合体进行比较。提案的最后部分(具体目标4)将研究多囊蛋白在维持细胞内钙稳态中的作用，并分析PKD1/2突变的后果。总体而言，这些研究的结果应该能更清楚地了解VSMCs中多囊蛋白的功能，以及这些蛋白的缺失在ADPKD相关的血管异常和高血压中可能发挥的作用。这一信息将为设计合理的治疗这些并发症的方法提供依据。这段时期的实验室学习，加上正式的职业发展计划，旨在将候选人培养为在ADPKD血管方面拥有独特专业知识的内科科学家。

DESCRIPTION (provided by applicant): Dr. Qian's long-term objective is to contribute to a better understanding of kidney disease pathophysiology, especially the vascular complications of autosomal dominant polycystic kidney disease (ADPKD). Her aim, over five-years, is to become a well-trained independent physician scientist with advance theoretical and technical knowledge in genetics and molecular and cellular biology through the implementation of this proposal. The candidate's mentor, Dr. Torres, is an expert in clinical and vascular aspects of ADPKD and her co-mentor, Dr. Harris, brings a detailed knowledge of ADPKD genetics. The third mentor, Dr. Farrugia, is an expert in Ca2+ homeostasis and will provide valuable guidance for that part of the proposal. The particular focus of this proposal is to study the role of the ADPKD proteins, polycystin-1 and -2 in the vasculature and to determine if defects in these proteins are directly related to the vascular manifestations and hypertension associated with ADPKD. Preliminary studies have indicated that the polycystins are expressed in vascular smooth muscle cells (VSMC) and that they interact as a part of a polycystin complex that may play a role in maintaining intracellular Ca2+ The first two Specific Aims of the proposal are to examine the consequences of mutation to the murine Pkdl and Pkd2 genes, and over expression of the human protein in mouse, on the characteristics of VSMCs. These will be analyzed in vitro, as cultured VSMCs and in vivo, by study of the ultrastructure of the vasculature in the mouse mutants. Specific Aim 3 will identify components of a VSMC polycystin complex using imaging methods, co-immunoprecipitation and the yeast two- hybrid technique. The results of these studies will allow a VSMC polycystin complex to be defined and comparison made to corresponding complexes in epithelial cells. The final part of the proposal (Specific Aim 4) will examine the role of the polycystins in maintaining intracellular calcium homeostasis and analyze the consequences of Pkd1/2 mutation. Overall, the results from these studies should provide a clearer view of the function of the polycystins in VSMCs and the likely role that loss of these proteins may play in the vascular abnormalities and hypertension associated with ADPKD. This information will provide the basis for designing rational therapies for the treatment of these complications. This period of laboratory based study, plus the formalized career development program, is designed to train the candidate as a physician scientist with a unique expertise in vascular aspects of ADPKD.