Role of Peroxidasin in Glomerular Matrix Homeostasis

过氧化物酶在肾小球基质稳态中的作用

基本信息

批准号：
9265081
负责人：
Gautam Bhave
金额：
$ 15.85万
依托单位：
VANDERBILT UNIVERSITY MEDICAL CENTER
依托单位国家：
美国
项目类别：
财政年份：
2013
资助国家：
美国
起止时间：
2013-07-15 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9265081
关键词：
Academic Medical Centers Acids Affect Afferent Neurons Apoptosis Atherosclerosis Attention Basement membrane Binding Biochemical Biochemistry Biological Assay Biology Career Choice Cell Adhesion Cell Communication Cell Surface Proteins Cell physiology Cells Chemicals Clinical Collagen Type IV Defect Development Diabetes Mellitus Diabetic Nephropathy Disease Doctor of Medicine Doctor of Philosophy End stage renal failure Environment Enzymes Epithelial Cells Exhibits Extracellular Matrix Fibrosis Functional disorder Funding Generations Glucose Goodpasture antigen Growth Factor Head Homeostasis Hospitals Hydrogen Peroxide Hyperglycemia Hypertension Hypochlorous Acid Immunoglobulin Domain Immunoglobulins Inflammation Injury Integrins Intervention Investigation Kidney Kidney Diseases Knowledge Laboratories Lead Leucine-Rich Repeat Lipids Manuscripts Mass Spectrum Analysis Mediating Medicine Mentors Modeling Mus Nature Nephrology Neurons Neurosciences Oxidants Pathogenesis Pathogenicity Pathologic Patients Peroxidases Pharmacology Phenotype Phloroglucinol Play Post-Translational Protein Processing Production Protein Binding Domain Proteins Publications Published Comment Publishing Pyridoxamine Reactive Oxygen Species Renal function Research Role Scientist Seminal Signal Transduction Site Superoxides Tertiary Protein Structure Testing Time Tissues Training Translating Tubular formation Tyrosine United States National Institutes of Health Up-Regulation Wit Work base career development college crosslink db/db mouse diabetic experience glomerular basement membrane glycation halogenation improved in vitro Model in vivo inhibitor/antagonist innovation insight instructor interstitial kidney cell mouse model mutant novel novel therapeutics oxidative damage peroxidasin podocyte public health relevance therapeutic target von Willebrand Factor

项目摘要

DESCRIPTION (provided by applicant): Diabetic nephropathy (DN) is the leading cause of end-stage renal disease in the U.S. A central derangement in diabetic nephropathy is excessive generation of reactive oxygen species (ROS). Superoxide and its product, hydrogen peroxide (H2O2), are ROS that have garnered greatest attention in DN. We recently discovered that peroxidasin, a protein embedded in extracellular matrix, converts H2O2 to highly reactive hypobromous (HOBr) and hypochlorous (HOCl) acid, collectively denoted as hypohalous acids (HOX). Peroxidasin uses HOX to form novel sulfilimine bonds (S=N) in basement membrane collagen IV, the first identified function for peroxidasin. But, in a diabetic milieu saturated wit H2O2 substrate, peroxidasin may produce excessive HOX. HOX are highly reactive and oxidatively halogenate proteins to adversely affect cell function. In preliminary work using a mouse model of diabetic nephropathy, we have found that glomerular and tubular peroxidasin expression and HOX mediated protein damage are increased. Based on these findings, we hypothesize that peroxidasin plays a pathogenic role in diabetic nephropathy by generating toxic HOX that halogenate proteins to alter cell function and promote extracellular matrix expansion. To test this hypothesis, Aim 1 will determine how peroxidasin functions within basement membranes examining the interaction between collagen IV and peroxidasin. In Aim 2, we will examine the role of peroxidasin and HOX in hyperglycemia induced renal cell dysfunction and matrix accumulation as an in vitro model of diabetic nephropathy. Using innovative mass spectrometry, we will identify specific sites of HOX mediated protein halogenation associated with peroxidasin upregulation. In Aim 3, we will directly test the role of peroxidasin and HOX in a mouse model of diabetic nephropathy. Building upon the mass spectrometry studies in Aim 2, we will identify protein halogenation sites in vivo. Furthermore, we will pharmacologically block peroxidasin and determine whether the intervention ameliorates diabetic nephropathy. The proposed work holds promise to provide insight into the pathogenesis of diabetic nephropathy and establish peroxidasin as a potential therapeutic target. Environment The proposed studies will be conducted at the Vanderbilt University Medical Center within the Division of Nephrology. The division has 14 NIH funded laboratories with over 13 million dollars in annual NIH funding and is one of only nine NIH George O'Brien Kidney Research Centers in the U.S. thus providing Dr. Bhave ample financial, administrative, and scientific support. Dr. Bhave is primarily mentored by Dr. Billy Hudson, a well- established scientist with nearly 40 years of NIH funding. His expertise resides in basement membrane and type IV collagen biochemistry and his seminal work includes the identification of the Goodpasture antigen and the development of pyridoxamine as a glycation inhibitor for the treatment of diabetic nephropathy. Dr. Bhave is also supported by Drs. Raymond Harris and Roy Zent as co-mentors to help develop an independent line of investigation and career path. Dr. Harris is Chief of the Division of Nephrology and his experience in models of diabetic nephropathy is crucial to the proposed work. Dr. Zent heads a well-funded group studying renal development and integrins and will provide expertise in the investigation of peroxidasin and cell-matrix interactions. Taken together, Dr. Bhave's mentors collectively represent over 75 years of experience in career development, mentoring, and scientific investigation. Candidate Dr. Gautam (Jay) Bhave is currently an Instructor in the Division of Nephrology at Vanderbilt with >80% of his time dedicated to bench research. He completed his M.D. and Ph.D. degrees at Baylor College of Medicine with High Honors and clinical training at Johns Hopkins Hospital and Vanderbilt. His graduate work examined sensitization of sensory neurons with tissue injury and inflammation and garnered high-profile, first author publications in Nature Neuroscience, Neuron, and PNAS. After clinical training, Dr. Bhave joined the lab of Dr. Billy Hudson as a research fellow and began investigating how a newly discovered sulfilimine (S=N) cross-link in the collagen IV protein network is formed. The work has reached fruition with the discovery of a catalyzing enzyme, peroxidasin, and a first author manuscript describing this work was recently published in Nature Chemical Biology accompanied by a commentary and cover illustration. Ultimately, Dr. Bhave hopes to translate his seminal biochemical discovery into an independent line of investigation examining the role of peroxidasin in matrix remodeling and expansion in renal disease.

描述（由申请人提供）：糖尿病性肾病（DN）是美国终末期肾脏疾病的主要原因，糖尿病性肾病中的中枢疾病是过量产生活性氧（ROS）。超氧化物及其产物，过氧化氢（H2O2）是在DN中获得最大关注的ROS。我们最近发现，过氧化物素是一种嵌入细胞外基质中的蛋白质，将H2O2转化为高反应性低含量（HOBR）和次氯酸盐（HOCL）酸，统称为低钙酸（HOX）。 Peroxidasin使用HOX在基底膜胶原蛋白IV中形成新型的硫亚胺键（S = N），这是过氧化物素的第一个鉴定功能。但是，在H2O2底物的糖尿病环境中，过氧化物素可能会产生过多的HOX。 HOX具有高反应性和氧化性卤代蛋白，可对细胞功能产生不利影响。在使用糖尿病肾病小鼠模型的初步工作中，我们发现肾小球和管状过氧化物素表达和HOX介导的蛋白质损伤增加了。基于这些发现，我们假设过氧化物素通过产生有毒Hox来在糖尿病性肾病中起致病作用，从而使卤代蛋白质以改变细胞功能并促进细胞外基质扩张。为了检验这一假设，AIM 1将确定在基底膜内的过氧化物素功能如何检查胶原蛋白IV和过氧化物素之间的相互作用。在AIM 2中，我们将检查过氧化物素和HOX在高血糖诱导的肾细胞功能障碍和基质积累中的作用，作为糖尿病性肾病的体外模型。使用创新的质谱法，我们将确定与过氧化物素上调有关的HOX介导的蛋白质卤素化的特定位点。在AIM 3中，我们将直接测试过氧化物素和HOX在A中的作用糖尿病肾病的小鼠模型。在AIM 2中的质谱研究基础上，我们将在体内鉴定蛋白质卤素化位点。此外，我们将在药理学上阻断过氧化物素，并确定干预措施是否可以改善糖尿病性肾病。提出的工作有望洞悉糖尿病性肾病的发病机理，并确立过氧化物素作为潜在的治疗靶点。环境该拟议的研究将在肾脏病科内的范德比尔特大学医学中心进行。该部门拥有14个NIH资助的实验室，拥有超过1300万美元的年度NIH资金，并且是美国仅有的9个NIH George O'Brien肾脏研究中心之一，因此为Bhave博士提供了丰富的财务，行政和科学支持。 Bhave博士主要由Billy Hudson博士指导，Billy Hudson博士拥有近40年的NIH资金。他的专业知识位于地下膜和IV型胶原蛋白生物化学，他的开创性工作包括鉴定良好的抗原和吡id胺作为治疗糖尿病性肾病的糖基化抑制剂的发展。 Bhave博士也得到了博士的支持。雷蒙德·哈里斯（Raymond Harris）和罗伊·Zent（Roy Zent）担任联合主管，以帮助发展独立的调查和职业道路。 Harris博士是肾脏病科主任，他在糖尿病肾病模型中的经验对于拟议的工作至关重要。 Zent博士负责研究肾脏发育和整联蛋白的资金丰富的小组，并将在研究过氧化物素和细胞 - 基质相互作用方面提供专业知识。综上所述，Bhave博士的导师共同代表了75年的职业发展，指导和科学研究经验。候选人Gautam博士（Jay）Bhave目前是Vanderbilt肾脏科学系的讲师，> 80％的时间致力于板凳研究。他完成了医学博士学位和博士学位约翰·霍普金斯医院和范德比尔特（Johns Hopkins Hospital）和范德比尔特（Vanderbilt）的贝勒医学院学位，获得高荣誉和临床培训。他的研究生工作检查了对感官神经元的组织损伤和炎症的敏感性，并获得了引人注目的自然神经科学，神经元和PNA中的第一作者出版物。经过临床培训，Bhave博士加入了Billy Hudson博士的实验室研究员，并开始研究胶原蛋白IV蛋白网络中新发现的硫胺（S = N）交联。由于发现了一种催化酶，过氧化物素的催化酶，这项工作已经实现了，描述这项工作的第一作者手稿最近发表在《自然化学生物学》中，并附有评论和封面插图。最终，Bhave博士希望将他的精确生化发现转化为独立的研究系列，研究过氧化物素在基质重塑和肾脏疾病中的扩张中的作用。