TRPC6 Regulation and its Role in Glomerular Pathology

描述(由申请人提供)： 研究发现，非特异性阳离子通道TRPC6的突变可导致遗传性局灶性节段性肾小球硬化。本项目旨在明确TRPC6在正常和异常肾小球功能中的作用，以确定TRPC6影响肾小球功能的关键信号通路。为此，候选人将：1)描述疾病相关突变子集增强TRPC6通道活性的机制；2)评估肾小球内各种细胞类型中TRPC6功能异常的相对重要性；以及3)确定NFAT转录因子家族作为肾小球TRPC6信号转导靶点的潜在作用。应聘者过去在许多基本的细胞生物学方法方面都有经验。此外，在Pollak博士实验室的博士后培训期间，他已经开始了解研究通道生物学、转录激活和开发肾脏疾病的尿液模型所需的技术和实验设计。他已经生成了相当多与该项目相关的初步数据。在他计划的项目期间，SchLondorff博士将能够掌握几项新技术，包括细胞内钙成像方法、Cre-Lox转基因动物生成、小鼠肾脏病理分析、免疫组织化学和微阵列基因表达技术。除了Pollak实验室的熟练成员，施隆多夫博士还获得了哈佛医学院几位有成就的专家的合作，以帮助他在这些领域的发展。 公共卫生相关性：局灶性节段性肾小球硬化(FSGS)是一种见于多种疾病状态的肾脏病变，在需要透析或肾移植的终末期肾病患者中占很大比例，目前缺乏有效的治疗方法。通过更好地了解离子通道中的遗传缺陷是如何导致这种疾病的，该项目希望确定关键过程，这些过程的中断会导致多种形式的肾脏疾病的FSGS和肾功能障碍。我们的目标是，这些见解将确定新的潜在目标，针对这些目标制定治疗干预措施。

DESCRIPTION (provided by applicant): Mutations in TRPC6, a nonspecific cation channel, have been found to lead to hereditary forms of focal segmental glomerulosclerosis. This project aims to define the role of TRPC6 in normal and abnormal glomerular function, with the goal of identifying the critical signaling pathways influenced by TRPC6. To this end, the candidate will: 1) characterize the mechanism whereby a subset of disease-associated mutations enhance TRPC6 channel activity; 2) assess the relative importance of abnormal TRPC6 function within various cell types within the glomerulus; and 3) establish the potential role of the NFAT transcription factor family as a target of TRPC6 signaling in the glomerulus. The candidate has past experience in a host of basic cell biology methods. In addition, during his post-doctoral fellowship training in Dr. Pollak's lab he has begun to develop an understanding of the techniques and experimental design required for studying channel biology, transcriptional activation, and for developing urine models of kidney disease. He has generated considerable preliminary data pertinent to the project. During his proposed project period, Dr. Schlondorff will be able to master several new techniques, including methods of intracellular calcium imaging, Cre-Lox transgenic animal generation, analysis of murine renal pathology, immunohistochemistry, and microarray gene expression technology. In addition to skilled members of the Pollak laboratory, Dr. Schlondorff has garnered the cooperation of several accomplished experts at the Harvard Medical School to assist him in these areas of development. PUBLIC HEALTH RELEVANCE: Focal segmental glomerular sclerosis (FSGS) is a kidney lesion seen in a number of disease states, contributes significantly to the population of patients with end-stage renal disease requiring dialysis or kidney transplantation, and currently lacks effective therapies. By gaining a better understanding of how genetic defects in an ion channel can lead to this disease, this project hopes to identify critical processes whose disruption lead to FSGS and kidney dysfunction in multiple forms of kidney disease. Our goal is that these insights will identify novel potential targets against which to develop therapeutic interventions.