Mesonephric Model of Podocyte Injury and Regeneration

摘要 本K99/R 00应用程序的目的是利用我开发的一种新的动物模型系统， 肾病综合征与足细胞再生的分子机制研究。这将有助于 我的研究生涯向独立调查员的位置过渡。 肾病综合征（NS）是一种以蛋白尿、低白蛋白血症、水肿和高蛋白血症为特征的肾脏疾病。 高脂血症，由于肾小球滤过屏障（GFB）功能的破坏。过度的原因 75%的激素抵抗型肾病综合征（SRNS）患者至今仍不明原因，无有效治疗方法 可用于该疾病，其导致局灶节段性肾小球硬化（FSGS）并导致 终末期肾病（ESKD）。SRNS遗传病因的新发现及啮齿类动物的研究 足细胞损伤模型揭示了足细胞在GFB的正常功能中的关键作用， NS的发病机制。然而，NS的啮齿动物模型不适合高通量筛选 用于该疾病的治疗剂。最近的研究已经确定了肾祖细胞的一个子集， 成年人肾脏能够再生肾上皮细胞，包括足细胞，这表明 新的干/祖细胞为基础的方法治疗肾病综合征。然而， 足细胞的再生机制 为了解决这些问题，我建议建立斑马鱼中肾作为一个新的模式系统， 肾病综合征：（1）利用转基因斑马鱼模型，我已经产生了筛选 改善蛋白尿的化学物质;（2）研究表达wt 1b的肾祖细胞的潜力 细胞再生斑马鱼足细胞及足细胞再生过程中基因表达的研究 已建立的GFB试验，用于筛选温度敏感性肾病斑马鱼突变体， 确定NS的新遗传原因。研究成果将提供（1）新的 化学化合物，可以导致治疗NS;（2）新的见解足细胞的机制， 再生;（3）参与NS发病机制的新基因;（4）新的斑马鱼模型， 疾病 作为儿科和传染病系的博士后研究员， 密歇根大学，我致力于开发斑马鱼中肾成为一种新的动物模型系统， 肾病综合症作为我的职业目标我的长期职业目标是成为一名 肾脏研究领域的独立研究者，专注于肾脏疾病的动物模型。的 该奖项的培训（K99）阶段将由Friedhelm Hildebrandt博士指导，他是 霍华德休斯医学研究所和国际公认的人类遗传学领域的领导者， 小儿肾脏疾病，包括肾病综合征。密歇根大学有几十个活跃的， 肾脏发育、肾脏疾病和斑马鱼发育的合作研究小组 和遗传学，这为我的培训和职业发展提供了理想的环境。

ABSTRACT The objective of this K99/R00 application is to utilize a new animal model system that I have developed for the study of molecular mechanisms of nephrotic syndrome and podocyte regeneration. This will aid the transition of my research career toward an independent investigator position. Nephrotic syndrome (NS) is a kidney disease characterized by proteinuria, hypoalbuminemia, edema, and hyperlipidemia, due to the disruption of renal glomerular filtration barrier (GFB) function. The causes of over 75% of steroid resistant nephrotic syndrome (SRNS) cases are still unknown and no effective therapy is available for the disease, which results in focal segmental glomerular sclerosis (FSGS) and leads to end-stage kidney disease (ESKD). Recent discoveries of genetic causes of SRNS and studies of rodent models of podocyte damage revealed the pivotal role of podocytes in the normal function of GFB and the pathogenesis of NS. However, rodent models for NS are not suitable for high-throughput screening of therapeutical agents for the disease. Recent studies have identified a subset of renal progenitor cells in adult human kidney that are capable of regenerating renal epithelial cells, including podocytes, suggesting a novel stem/progenitor cell-based approach for the treatment of nephrotic syndrome. However, little is known about the regenerative mechanism of podocytes. To address these questions, I propose to establish zebrafish mesonephros as a new model system for nephrotic syndrome by: (1) utilizing the transgenic zebrafish models that I have generated to screen for chemicals ameliorating proteinuria; (2) investigating the potential of wt1b-expressing renal progenitor cell to regenerate podocytes in zebrafish and gene expression during podocyte regeneration; (3) using the established GFB assay(s) to screen for temperature-sensitive nephrotic zebrafish mutants and identify novel genetic causes of NS. Accomplishment of the proposed research will provide (1) new chemical compounds that can lead to therapies for NS; (2) new insights into the mechanism of podocyte regeneration; (3) new genes that are involved in the pathogenesis of NS; and (4) new zebrafish models for the disease. Being a postdoctoral research fellow in the Department of Pediatrics and Communicable Diseases, University of Michigan, I am committed to develop zebrafish mesonephros into a new animal model system for nephrotic syndrome as my immediate career goal. My long-term career goal is to establish myself as an independent investigator in the field of kidney research, focusing on animal models of kidney diseases. The training (K99) phase of this award will be mentored by Dr. Friedhelm Hildebrandt, who is an investigator of Howard Hughes Medical Institute and internationally recognized leader in the fields of human genetics of pediatric kidney diseases, including nephrotic syndrome. University of Michigan has dozens of active and collaborative research groups working on kidney development, kidney diseases, and zebrafish development and genetics, which provides an ideal environment for my training and career development.