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：
8700396
负责人：
Gautam Bhave
金额：
$ 14.51万
依托单位：
VANDERBILT UNIVERSITY
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/8700396
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 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 Pathologic Patients Peroxidases Phenotype Phloroglucinol Play Post-Translational Protein Processing Production Protein Binding Domain Proteins Publications Published Comment Publishing Pyridoxamine Reactive Oxygen Species Renal Hypertension 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 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）：糖尿病肾病（DN）是美国终末期肾病的主要原因。糖尿病肾病的核心紊乱是活性氧（ROS）的过度产生。超氧化物及其产物过氧化氢 (H2O2) 是 DN 中最受关注的 ROS。我们最近发现过氧化物酶（一种嵌入细胞外基质中的蛋白质）可将 H2O2 转化为高反应性次溴酸 (HOBr) 和次氯酸 (HOCl)，统称为次卤酸 (HOX)。过氧化物酶利用 HOX 在基底膜胶原 IV 中形成新型硫亚胺键 (S=N)，这是过氧化物酶第一个确定的功能。但是，在 H2O2 底物饱和的糖尿病环境中，过氧化物酶可能会产生过量的 HOX。 HOX 是高反应性和氧化卤化蛋白，会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，我们发现肾小球和肾小管过氧化物酶表达以及 HOX 介导的蛋白质损伤增加。基于这些发现，我们假设过氧化物酶通过产生有毒的 HOX 来卤化蛋白质以改变细胞功能并促进细胞外基质扩张，从而在糖尿病肾病中发挥致病作用。为了检验这一假设，目标 1 将确定过氧化物酶在基底膜内的功能，检查 IV 型胶原蛋白和过氧化物酶之间的相互作用。在目标 2 中，我们将作为糖尿病肾病的体外模型，研究过氧化物酶和 HOX 在高血糖诱导的肾细胞功能障碍和基质积累中的作用。使用创新的质谱分析法，我们将确定 HOX 介导的与过氧化物酶上调相关的蛋白质卤化的特定位点。在目标 3 中，我们将直接测试过氧化物酶和 HOX 在糖尿病肾病小鼠模型。基于目标 2 中的质谱研究，我们将鉴定体内蛋白质卤化位点。此外，我们将通过药物阻断过氧化物酶并确定干预措施是否可以改善糖尿病肾病。这项工作有望深入了解糖尿病肾病的发病机制，并将过氧化物酶确立为潜在的治疗靶点。环境拟议的研究将在范德比尔特大学医学中心肾脏病科进行。该部门拥有 14 个 NIH 资助的实验室，每年 NIH 资助超过 1300 万美元，是美国仅有的 9 个 NIH 乔治·奥布莱恩肾脏研究中心之一，从而为 Bhave 博士提供了充足的财务、行政和科学支持。 Bhave 博士主要受到 Billy Hudson 博士的指导，Billy Hudson 博士是一位知名科学家，获得 NIH 近 40 年的资助。他的专业知识在于基底膜和 IV 型胶原生物化学，他的开创性工作包括 Goodpasture 抗原的鉴定以及开发吡哆胺作为治疗糖尿病肾病的糖化抑制剂。 Bhave 博士也得到了 Drs. 的支持。雷蒙德·哈里斯 (Raymond Harris) 和罗伊·赞特 (Roy Zent) 作为共同导师，帮助制定独立的调查路线和职业道路。 Harris 博士是肾病科主任，他在糖尿病肾病模型方面的经验对于拟议的工作至关重要。 Zent 博士领导着一个资金充足的肾脏发育和整合素研究小组，并将提供过氧化物酶和细胞基质相互作用研究方面的专业知识。总的来说，Bhave 博士的导师在职业发展、指导和科学研究方面总共拥有超过 75 年的经验。候选人 Gautam (Jay) Bhave 博士目前是范德堡大学肾脏科的讲师，他 80% 以上的时间致力于实验室研究。他完成了医学博士和博士学位。在贝勒医学院获得最高荣誉学位，并在约翰·霍普金斯医院和范德比尔特接受临床培训。他的研究生工作研究了感觉神经元对组织损伤和炎症的敏感性，并在《Nature Neuroscience》、《Neuron》和《PNAS》上发表了备受瞩目的第一作者论文。经过临床培训后，Bhave 博士加入 Billy Hudson 博士的实验室担任研究员，开始研究新发现的 IV 型胶原蛋白网络中的硫亚胺 (S=N) 交联是如何形成的。随着催化酶过氧化酶的发现，这项工作已经取得了成果，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》上，并附有评论和封面插图。最终，Bhave 博士希望将他的开创性生化发现转化为独立的研究路线，检查过氧化物酶在肾脏疾病基质重塑和扩张中的作用。