The Role of Aminopeptidase-A in GBM Thickening in Diabetic Nephropathy

氨肽酶-A 在糖尿病肾病 GBM 增厚中的作用

基本信息

批准号：
9378083
负责人：
Caroline B Marshall
金额：
--
依托单位：
BIRMINGHAM VA 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：
2013-10-01 至 2018-09-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/9378083
关键词：
Address Affect Albuminuria Applications Grants Award BALB/cJ Mouse Binding Biological Breeding Cell Nucleus Chronic Kidney Failure Clinical Communication Culture Media Cytoplasmic Tail Development Development Plans Diabetes Mellitus Diabetic Nephropathy Disease Dissociation Down-Regulation End stage renal failure Epithelial Cells Expression Profiling Extracellular Matrix Family Funding Gene Expression Gene Targeting Genes Glucose Goals Grant Harvest Heterodimerization Human Hyperglycemia Image In Vitro Injury Insulin Insulin Receptor Integral Membrane Protein Kidney Knock-out Knockout Mice Laboratories Link Manuscripts Measurement Mediating Mentors Messenger RNA Modeling Molecular Morphology Mus Nuclear Nuclear Localization Signal Nuclear Translocation Pathogenesis Pathway interactions Patients Phenotype Play Population Prevalence Proteins Publishing Rattus Receptor Down-Regulation Receptor Signaling Regulation Renin-Angiotensin System Reporting Research Role Schedule Small Interfering RNA Streptozocin Testing Thick TimeLine Training United States United States National Institutes of Health Western Blotting Work Writing Zinc Fingers base career development clinical care db/db mouse design diabetic diabetic patient diabetic rat expectation glomerular basement membrane glutamyl aminopeptidase improved in vivo knock-down meetings member migration overexpression patient population podocyte protein protein interaction public health relevance skills targeted treatment transcription factor type I diabetic

项目摘要

DESCRIPTION (provided by applicant): PROJECT SUMMARY Diabetes mellitus is the most common cause of chronic kidney disease. Approximately 40% of diabetic patients develop diabetic nephropathy (DN); and nearly half of all incident cases of end-stage renal disease in the United States are due to DN. Despite widespread use of targeted therapies to lower glucose and to antagonize the renin-angiotensin system, the prevalence of DN has remained stable. Thus, effective and disease-specific therapies for DN are needed. The immediate goal of the applicant is to study the mechanisms underlying the development of glomerular basement membrane (GBM) thickening in DN. The applicant's long- term goal is to understand the pathogenesis of diabetic kidney disease in sufficient detail to develop mechanism-based therapies for DN. A clear and focused career development plan has been designed by the applicant and the mentoring team to provide new and enhanced training in cell-extracellular matrix interactions in DN, insulin receptor signaling, communication and presentation skills, grant and manuscript writing, and responsible conduct in research. There is a plan for progressive increase in independence over the course of the CDA-2 Award. Timelines for the specific aims, laboratory space allocation, scheduled meetings between the applicant and the mentoring team, expectations of the applicant, and the specific role(s) of each mentor are addressed in detail in the career development plan. In addition, the applicant is expected to submit smaller grant proposals in the earlier years and an application for independent funding (VA Merit Award, NIH R01) in the last two years of the CDA-2 Award. Studies currently being conducted by the applicant have demonstrated that aminopeptidase-A (APA), a type II transmembrane protein expressed in visceral glomerular epithelial cells (podocytes), may play an important role in the pathogenesis of GBM thickening in DN. Initial results show that APA knockout mice have significantly thickened GBMs. Moreover, APA is down-regulated in patients with DN and in rat models of type 2 DN. Also, knockdown of the insulin receptor in cultured glomerular epithelial cells reduces the expression of APA. In this proposal, the applicant will study the role of APA downregulation in mediating the expression of genes involved in GBM remodeling. The overall hypothesis is that APA down-regulation in the diabetic state causes displacement of transcriptional factors (ZHX proteins) that migrate into the podocyte nucleus and induce changes in the expression of matrix- related genes, thereby leading to GBM thickening. The goal of the proposed studies is to increase our understanding of key molecular changes that result in the development of GBM thickening and DN. In Specific Aim 1, the effects of APA down-regulation on matrix-related gene expression will be studied. In Specific Aim 2, the importance of APA-ZHX interaction in the development of GBM thickening will be examined. In Specific Aim 3, the effects of insulin receptor deficiency on APA and ZHX proteins will be investigated.

描述（由申请人提供）：项目摘要糖尿病是慢性肾脏疾病的最常见原因。大约40％的糖尿病患者患有糖尿病性肾病（DN）；在美国，几乎一半的终末期肾脏疾病发生的事件是由于DN引起的。尽管广泛使用靶向疗法来降低葡萄糖并拮抗肾素 - 血管紧张素系统，但DN的患病率仍然稳定。因此，需要进行有效的DN和特异性疗法。申请人的直接目标是研究DN中肾小球基底膜（GBM）增厚的基础机制。申请人的长期目标是了解糖尿病肾脏疾病的发病机理，足以开发基于机制的DN疗法。申请人和指导团队设计了一项清晰且重点的职业发展计划，以在DN，胰岛素受体信号传导，沟通和表现技巧，赠款和手稿写作以及负责任的研究中提供新的和增强的培训。在CDA-2奖的过程中，有一个计划逐步提高独立性的计划。在职业发展计划中详细介绍了具体目标的时间表，实验室空间分配，申请人与指导团队之间的预定会议，申请人的期望以及每个导师的特定角色。此外，预计申请人将在较早的几年中提交较小的赠款建议，并在CDA-2奖的最后两年中提交独立资金申请（VA MEARIT Award，NIH R01）。申请人目前正在进行的研究表明，在内脏肾小球上皮细胞（足细胞）中表达的II型跨膜蛋白（APA）在DN中GBM增厚的发病机理中可能起重要作用。初始结果表明，APA敲除小鼠的GBM显着增厚。此外，DN患者和2型DN的大鼠模型中APA被下调。同样，培养的肾小球上皮细胞中胰岛素受体的敲低也降低了APA的表达。在此提案中，申请人将研究APA下调在介导GBM重塑涉及的基因表达中的作用。总体假设是，糖尿病状态的APA下调会导致转录因子（ZHX蛋白）的位移，该因子迁移到足细胞核中，并诱导与基质相关基因表达的变化，从而导致GBM增厚。拟议研究的目的是增加我们对关键分子变化的理解，从而导致GBM增厚和DN的发展。在特定的目标1中，将研究APA下调对基质相关基因表达的影响。在特定的目标2中，将检查APA-ZHX相互作用在GBM增厚开发中的重要性。在特定的目标3中，将研究胰岛素受体缺乏症对APA和ZHX蛋白的影响。