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: 8606460
• 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/8606460
• 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. PUBLIC HEALTH RELEVANCE: Chronic and end-stage kidney disease (CKD and ESKD) results in significant morbidity and mortality. We propose to examine novel pathways that may provide insight into mechanisms of proteinuric kidney disease and focal glomerulosclerosis (FSGS), a leading cause of childhood CKD and ESKD. We will examine the role of Par1a/b in establishing and maintaining podocyte structure and define polarity protein expression in human nephrotic syndrome and glomerulosclerosis, potentialy identifying new targets for therapeutic intervention or biomarkers for progressive kidney disease. !

描述（由申请人提供）：该提案描述了足细胞生物学和肾小球疾病学术生涯发​​展的五年计划。 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。该计划将促进利用分子遗传学和细胞生物学的专业知识来研究在肾小球疾病中维持足细胞极性的机制。博士。 Katalin Susztak 和 Anne Muesch 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，也是该领域识别肾小球疾病新机制的领导者。内部共同导师 Muesch 博士是发育和分子生物学副教授，他是建立细胞极性机制的专家，该机制将在肾小球疾病的背景下进行检查。此外，为了扩展 PI 的技能并促进她对足细胞极性发展的研究，PI 正在接受来自附近哥伦比亚大学的外部顾问 Jonathan Barasch 博士的胚胎肾培养实践培训。博士。 Susztak、Muesch、Barasch 和其他几位备受推崇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。研究将重点关注顶端-基底极性蛋白 Par1a/b 在肾脏发育过程中建立足细胞极性以及在肾小球疾病中维持足细胞极性中的作用。最近在她导师的指导下进行的工作表明，Par1a/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，Par1a/b 功能的显性负性抑制会引起细胞形状的变化和缝隙隔膜蛋白表达的改变，缝隙隔膜蛋白是支持肾小球滤过器结构和功能的关键足细胞成分。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 Par1a/b 表达发生改变。拟议的实验将需要使用体内小鼠模型对 Par1a/b 功能进行多西环素诱导的足细胞特异性抑制，这将允许研究肾单位发育（胚胎发生过程中的诱导）期间和成年小鼠中的 Par1a/b 功能。在胚胎肾培养物中抑制 Par1a/b 功能的构建体的腺病毒感染将用于进一步研究 Par1a/b 的体外功能。此外，使用嘌呤霉素氨基核苷肾病 (PAN) 诱导大鼠肾病综合征和肾小球硬化实验模型中的 Par1a/b 表达，以及使用对照和患病人类肾脏标本的集合在人类肾病综合征中的表达。具体目标包括：检查肾单位发育过程中 Par1a/b 对足细胞分化的功能，2) 检查 Par1a/b 在维持极化足细胞结构和肾小球滤过功能方面的功能，以及 3) 检查 Par1a/b 在肾病综合征和肾小球硬化的人和实验模型中的表达。 公共卫生相关性：慢性和终末期肾病（CKD 和 ESKD）会导致显着的发病率和死亡率。我们建议研究新的途径，以深入了解蛋白尿肾病和局灶性肾小球硬化症 (FSGS) 的机制，FSGS 是儿童 CKD 和 ESKD 的主要原因。我们将研究 Par1a/b 在建立和维持足细胞结构中的作用，并定义人类肾病综合征和肾小球硬化症中极性蛋白的表达，从而有可能确定治疗干预的新靶标或进行性肾脏疾病的生物标志物。 ！