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）：该提案描述了一项为期五年的足够细胞生物学和肾小球疾病的学术生涯的计划。 PI已经完成了一项由NIH支持的，结构化的奖学金培训计划，用于儿科和发育肾脏科，现在将通过独特的跨部门资源整合来扩展她的科学技能。该计划将促进使用分子遗传学和细胞生物学的专业知识，以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。她的主要导师苏兹塔克（Susztak）博士是内科/肾脏学的副教授，也是她在识别肾小球疾病的新机制方面的领导者。内部联合学者Muesch博士是开发和分子生物学副教授，是建立细胞极性的机制专家，将在肾小球疾病的背景下进行检查。此外，为了扩大PI的曲目，并促进了她对足细胞极性发展的研究，PI正在接受外部顾问Jonathan Barasch博士在哥伦比亚大学附近接受外部顾问Jonathan Barasch博士接受胚胎肾脏文化的动手培训。博士。 Susztak，Muesch，Barasch和其他几位备受推崇的发展肾病学家将组成一个咨询委员会，以提供科学和职业建议。研究将侧重于顶端极性蛋白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方面的作用。在她的导师的指导下进行的最近的工作表明，par1a/b在发展肾上腺和啮齿动物和人类足细胞中表达。培养的足细胞中PAR1A/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，它们是支持肾小球滤波器结构和功能的关键足细胞成分。在糖尿病性肾病，肾病综合征和肾小球硬化的啮齿动物模型中，肾小球par1a/b的表达改变了。提出的实验将使用体内小鼠模型对强力霉素诱导的，足细胞特异性的par1a/b功能抑制，这将允许研究肾单位发育期间的PAR1A/B功能（在依次生成期间的诱导）和成年小鼠和成年小鼠。在胚胎肾脏培养中抑制PAR1A/B功能的构建体的腺病毒感染将用于进一步研究PAR1A/B体外的功能。此外，使用毒素综合征和肾小球硬化的实验模型中的PAR1A/B表达，使用毒素氨基核苷肾脏病（PAN）在大鼠和人类肾病综合征中诱导的，使用控制和患病的人类肾脏肾脏标本的集合。具体目的包括：检查肾脏分化的肾单位发育过程中的par1a/b功能，2）检查par1a/b的功能，以维持偏光性足细胞结构和肾小球滤波器功能，以及3）检查par1a/b在人类和实验模型的肾病综合征和胶质膜疾病中的par1a/b的表达。