Kidney disease mechanisms associated with human genetic variation

与人类遗传变异相关的肾脏疾病机制

基本信息

批准号：
8642932
负责人：
Leslie A Bruggeman
金额：
$ 54.27万
依托单位：
CASE WESTERN RESERVE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-07-20 至 2018-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8642932
关键词：
AIDS-Associated Nephropathy Accounting Address African African American American Animals Apolipoproteins Autophagocytosis Autophagosome Binding Biological Biology Biopsy Specimen Blood Cardiovascular Diseases Cell Culture Techniques Cell Line Cells Cessation of life Chromosomes, Human, Pair 22 Chronic Kidney Failure Code Complementary DNA Complex Data Development Diagnosis Disease Docking Electron Microscopy End stage renal failure Environmental Risk Factor Expenditure Experimental Designs Gene Expression Profile Genes Genetic Epistasis Genetic Predisposition to Disease Genetic Variation Genotype Goals HIV HIV Infections HIV-1 Haplotypes Healthcare Home environment Homeostasis Human Human Genetics Immunofluorescence Immunologic In Situ In Situ Hybridization In Vitro Infection Infection prevention Injury Kidney Kidney Diseases Kidney Failure Knowledge Location Lysosomes Maps Mediating Medicare Metabolic Methods Modeling Mus NPHS2 protein Normal tissue morphology Pathogenesis Pathology Pathway interactions Patients Pattern Phenotype Population Prevention therapy Protein Biosynthesis Proteins Publishing Regulation Resistance Risk SNAP receptor Severities Signal Transduction Stress Testing Time Tissues Transgenic Mice Trypanosoma United States Variant Viral Viral Proteins Virus base diabetic disease diagnosis disease phenotype genetic association health disparity high risk improved in vivo in vivo Model mouse model nef Protein nephrin non-diabetic novel overexpression particle pathogen podocyte promoter public health relevance research study response risk variant transgene expression uptake vector

项目摘要

Chronic kidney disease (CKD), like many complex disease phenotypes, reflects dependent interactions between genetic susceptibilities and environmental factors. APOL1 variants explain much of the excess risk of advanced non-diabetic CKD in patients of African ancestry. However, only some patients with risk genotypes develop kidney disease, suggesting that genetic epistasis or environmental stress is required to trigger variant APOL1-dependent kidney injury. To investigate how APOL1 variants result in CKD, we have focused on HIV- associated nephropathy (HIVAN), the disease most robustly associated with APOL1 risk haplotypes and clearly dependent on an environmental factor, HIV-1 infection. Our preliminary data demonstrate APOL1 is expressed in the podocyte, and APOL1 overexpression activates autophagy. Although mTor-dependent suppression of core autophagy may cause diabetic glomerulopathy, a renal pathology not associated with APOL1 variants, we provide evidence that the genetic association of APOL1 with CKD results from APOL1- dependent regulation of distinct, selective autophagy pathways, which result in the destruction of pathogens. We hypothesize that APOL1 is an autophagic adaptor that tethers a docking SNARE (VAMP8) displayed on lysosomes to the autophagosomal protein LC3-II. The tethering function of APOL1 is activated by binding a triggering molecule, the HIV protein Nef, and results in the selective degradation of autophagosomes carrying HIV viral particles and proteins. Variant APOL1 proteins are functionally defective, permitting HIV to persist in the infected cell and continue synthesis of viral proteins including Nef. Nef blocks autophagic elimination of the virus and is critical for development of HIVAN. Our proposed experiments address three unanswered and novel questions regarding APOL1 function in CKD. Is APOL1 synthesized in situ in kidney and what is its subcellular home? Does circulating or renal-expressed variant APOL1 mediate kidney disease? Does dysregulation of autophagy pathways activated in response to specific environmental stresses result in kidney disease in patients with APOL1 risk genotypes? These questions are addressed with the following Specific Aims: 1. Determine APOL1 expression and intracellular location in normal kidney, and determine if APOL1 localization varies with risk genotype and/or disease diagnosis; 2. Generate in vivo models for the study of APOL1 function; 3. Characterize protein interactions between APOL1 with VAMP8 and the HIV protein Nef and examine normal and variant APOL1 regulation of autophagy in cell lines and cultured podocytes. These studies can result in novel, mechanism-based therapies, improve health disparities in African American patients and identify novel mechanisms of human CKD.

慢性肾脏病 (CKD) 与许多复杂的疾病表型一样，反映了依赖性相互作用遗传易感性和环境因素之间的关系。 APOL1 变异在很大程度上解释了非洲血统患者的晚期非糖尿病 CKD。然而，只有部分具有风险基因型的患者发展为肾脏疾病，表明需要遗传上位或环境压力来触发变异 APOL1 依赖性肾损伤。为了研究 APOL1 变异如何导致 CKD，我们重点关注 HIV- 相关性肾病 (HIVAN)，这种疾病与 APOL1 风险单倍型密切相关，显然取决于环境因素，即 HIV-1 感染。我们的初步数据表明 APOL1 是在足细胞中表达，APOL1 过度表达会激活自噬。虽然依赖 mTor 核心自噬的抑制可能会导致糖尿病肾小球病，这是一种与糖尿病肾病无关的肾脏病理学 APOL1 变体，我们提供证据表明 APOL1 与 CKD 的遗传关联源自 APOL1- 不同的选择性自噬途径的依赖性调节，从而导致病原体的破坏。我们假设 APOL1 是一个自噬适配器，它连接显示在上的对接 SNARE (VAMP8) 溶酶体结合自噬体蛋白 LC3-II。 APOL1 的束缚功能是通过结合触发分子，HIV 蛋白 Nef，并导致携带的自噬体选择性降解 HIV 病毒颗粒和蛋白质。变体 APOL1 蛋白存在功能缺陷，使得 HIV 能够持续存在感染细胞并继续合成包括 Nef 在内的病毒蛋白。 Nef 阻断自噬消除病毒，对于 HIVAN 的发展至关重要。我们提出的实验解决了三个悬而未决的问题关于 CKD 中 APOL1 功能的新问题。 APOL1 是在肾脏中原位合成的吗？它的作用是什么？亚细胞之家？循环或肾脏表达的 APOL1 变异是否介导肾脏疾病？做响应特定环境压力而激活的自噬途径失调会导致肾脏损伤 APOL1 风险基因型患者会罹患疾病吗？这些问题通过以下具体内容得到解决目的： 1. 测定APOL1在正常肾脏中的表达及细胞内定位，并确定APOL1是否存在定位随风险基因型和/或疾病诊断而变化； 2. 生成用于研究的体内模型 APOL1功能； 3. 表征 APOL1 与 VAMP8 和 HIV 蛋白 Nef 之间的蛋白质相互作用检查细胞系和培养足细胞中正常和变异的 APOL1 对自噬的调节。这些研究可以产生新颖的、基于机制的疗法，改善非裔美国人的健康差异患者并确定人类 CKD 的新机制。