Vascular Complications of Polycystic Kidney Disease

多囊肾的血管并发症

• 批准号: 6891356
• 负责人: QI QIAN
• 金额: $ 11.98万
• 依托单位: MAYO CLINIC ROCHESTER
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-06-01 至 2008-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6891356
• 关键词: calcium channel; calcium flux; cardiovascular disorder; cardiovascular imaging /visualization; disease /disorder model; gene mutation; genetically modified animals; hypertension; immunoprecipitation; laboratory mouse; medical complication; molecular pathology; polycystic kidney; protein localization; protein structure function; proteins; tissue /cell culture; vascular smooth muscle; yeast two hybrid system

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.

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多囊肾病（ADPKD）的血管并发症。她的目标是在五年内通过实施该提案，成为一名训练有素的独立医师科学家，在遗传学、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，她的搭档 Harris 博士带来了 ADPKD 遗传学的详细知识。第三位导师 Farrugia 博士是 Ca2+ 稳态方面的专家，将为提案的这一部分提供宝贵的指导。该提案的特别重点是研究 ADPKD 蛋白、多囊蛋白-1 和 -2 在脉管系统中的作用，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，并且它们作为多囊蛋白复合物的一部分相互作用，可能在维持细胞内 Ca2+ 方面发挥作用。该提案的前两个具体目标是检查鼠 Pkd1 和 Pkd2 基因突变以及小鼠中人类蛋白过度表达对 VSMC 特征的影响。这些将在体外（培养的 VSMC）和体内（通过研究小鼠突变体的脉管系统超微结构）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 多囊蛋白复合物的成分。这些研究的结果将允许定义 VSMC 多囊蛋白复合物，并与上皮细胞中相应的复合物进行比较。该提案的最后部分（具体目标 4）将检查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可以更清楚地了解 VSMC 中多囊蛋白的功能，以及这些蛋白的丢失在与 ADPKD 相关的血管异常和高血压中可能发挥的作用。该信息将为设计治疗这些并发症的合理疗法提供基础。这一阶段的实验室研究加上正式的职业发展计划，旨在将候选人培养成为一名在 ADPKD 血管方面拥有独特专业知识的医师科学家。