Role of Par1 Polarity Proteins in Podocyte Development and Glomerular Disease

Par1 极性蛋白在足细胞发育和肾小球疾病中的作用

• 批准号: 8418732
• 负责人: Kimberly Jean Reidy
• 金额: $ 13.34万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-04-01 至 2016-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8418732
• 关键词: Actins; Address; Adult; Advisory Committees; Apical; Biological Markers; Biology; Biopsy; Cell Polarity; Cell Shape; Cell membrane; Cellular biology; Childhood; Chronic; Collection; Complex; Cytoskeleton; Development; Diabetic Nephropathy; Disease; Dominant-Negative Mutation; Doxycycline; Embryo; End stage renal failure; Epithelial Cells; Experimental Models; Family member; Fellowship; Focal Segmental Glomerulosclerosis; Focal glomerulosclerosis; Foot Process; Future; Human; Immunofluorescence Immunologic; In Vitro; Infection; Internal Medicine; Investigation; Kidney; Kidney Diseases; Maintenance; Mentors; Molecular Biology; Molecular Genetics; Morbidity - disease rate; Mus; Nephrology; Nephrons; Nephrosis; Nephrotic Syndrome; Pathway interactions; Patients; Phosphorylation; Program Development; Protein-Serine-Threonine Kinases; Proteins; Proteinuria; Puromycin Aminonucleoside; Rattus; Regulation; Renal glomerular disease; Research; Resources; Rodent; Rodent Model; Role; Shapes; Signal Transduction; Specimen; Structure; Therapeutic Intervention; Time; Training; Training Programs; United States National Institutes of Health; Universities; Work; career; glomerulosclerosis; in vivo; insight; mortality; mouse model; nephrin; nephrogenesis; novel; podocyte; professor; programs; protein expression; protein function; public health relevance; research study; skills; slit diaphragm; therapeutic target

DESCRIPTION (provided by applicant): This proposal describes a five year program for development of an academic career in podocyte biology and glomerular disease. The PI has completed an NIH-supported, structured fellowship training program in Pediatric and Developmental Nephrology and will now expand upon her scientific skills through a unique integration of interdepartmental resources. This program will promote expertise in use of molecular genetics and cell biology to investigate mechanisms that maintain podocyte polarity in the setting of glomerular disease. Drs. Katalin Susztak and Anne Muesch will mentor the PI's scientific development. Her primary mentor, Dr. Susztak, is an Associate Professor of Internal Medicine/ Nephrology and a leader in her field in identifying novel mechanisms of glomerular disease. The internal co-mentor, Dr. Muesch, an Associate Professor in Development and Molecular Biology, is an expert in mechanisms that establish cell polarity, which will be examined in the context of glomerular disease. In addition, to expand the PI's repertoire and facilitate her study of the development of podocyte polarity, the PI is receiving hands-on training in embryonic kidney culture from an external advisor, Dr. Jonathan Barasch, who is located nearby at Columbia University. Drs. Susztak, Muesch, Barasch and several other highly regarded developmental nephrologists will form an advisory committee to provide scientific and career advice. Research will focus on the role of apical-basal polarity proteins Par1a/b in establishing podocyte polarity during kidney development and in maintaining podocyte polarity in the setting of glomerular disease. Recent work performed under the guidance of her mentors demonstrated that Par1a/b is expressed in developing nephrons and in rodent and human podocytes. Dominant negative suppression of Par1a/b function in cultured podocytes induced changes of cell shape and altered expression of slit diaphragm proteins, which are key podocyte components that support glomerular filter structure and function. Glomerular Par1a/b expression was altered in rodent models of diabetic nephropathy, nephrotic syndrome and glomerulosclerosis. The proposed experiments will entail doxycycline-inducible, podocyte-specific suppression of Par1a/b function using an in vivo mouse model, which will allow study of Par1a/b function during nephron development (induction during embyrogenesis) and in adult mice. Adenoviral infection of constructs suppressing Par1a/b function in embryonic kidney culture will be used to further study the function of Par1a/b in vitro. In addition, Par1a/b expression in an experimental model of nephrotic syndrome and glomerulosclerosis, using puromycin aminonucleoside nephrosis (PAN) induced in rats, and in human nephrotic syndrome, using a collection of control and diseased human kidney specimens. Specific aims include: Examine Par1a/b function during nephron development on podocyte differentiation, 2) Examine Par1a/b function in maintaining a polarized podocyte structure and glomerular filter function, and 3) Examine expression of Par1a/b in human and experimental models of nephrotic syndrome and glomerulosclerosis.

描述（由申请人提供）：本提案描述了一个为期五年的计划，在足细胞生物学和肾小球疾病的学术生涯的发展。PI已经完成了NIH支持的儿科和发育肾病学结构化奖学金培训计划，现在将通过独特的跨部门资源整合来扩展她的科学技能。该项目将促进分子遗传学和细胞生物学的应用，以研究肾小球疾病中维持足细胞极性的机制。Katalin Susztak和Anne Muesch博士将指导PI的科学发展。她的主要导师Susztak博士是内科/肾病学副教授，也是她在确定肾小球疾病新机制领域的领导者。内部共同导师Muesch博士是发育和分子生物学副教授，是建立细胞极性机制的专家，这将在肾小球疾病的背景下进行检查。此外，为了扩大PI的技能并促进其对足细胞极性发育的研究，PI正在接受外部顾问Jonathan Barasch博士的胚胎肾培养实践培训，该顾问位于附近的哥伦比亚大学。Susztak博士、Muesch博士、Barasch博士和其他几位德高望重的发育肾病学家将组成一个咨询委员会，提供科学和职业建议。研究将集中在顶端-基底极性蛋白Par 1a/B在肾脏发育过程中建立足细胞极性和在肾小球疾病背景下维持足细胞极性的作用。最近在她的导师指导下进行的工作表明，Par 1a/B在发育中的肾单位以及啮齿动物和人类足细胞中表达。在培养的足细胞中，对Par 1a/B功能的显性负抑制诱导细胞形状的变化和狭缝隔膜蛋白的表达改变，狭缝隔膜蛋白是支持肾小球滤过器结构和功能的关键足细胞组分。在糖尿病肾病、肾病综合征和肾小球硬化的啮齿动物模型中，肾小球Par 1 a/B表达改变。所提出的实验将需要使用体内小鼠模型对Par 1a/B功能进行强力霉素诱导的足细胞特异性抑制，这将允许在肾单位发育期间（胚胎发生期间的诱导）和成年小鼠中研究Par 1a/B功能。腺病毒感染抑制胚胎肾培养中Par 1a/B功能的构建体将用于进一步研究Par 1a/B的体外功能。此外，在肾病综合征和肾小球硬化的实验模型中，使用嘌呤霉素氨基糖苷肾病（PAN）在大鼠中诱导Par 1a/B表达，以及在人肾病综合征中，使用对照和患病人肾标本的集合。具体目标包括：检查Par 1a/B在肾单位发育期间对足细胞分化的功能，2）检查Par 1a/B在维持极化足细胞结构和肾小球滤过功能中的功能，和3）检查Par 1a/B在肾病综合征和肾小球硬化症的人和实验模型中的表达。