Sidekick proteins in HIV-associated nephropathy

HIV 相关肾病中的 Sidekick 蛋白

• 批准号: 7252596
• 负责人: Lewis Kaufman
• 金额: $ 12.87万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-07-01 至 2010-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7252596
• 关键词: AIDS-Associated Nephropathy; Actins; Address; Adhesions; Affect; Architecture; Axon; Binding; Biopsy; Bundling; Cell Adhesion Molecules; Cells; Cessation of life; Chromosome Pairing; Cultured Cells; Cytoskeleton; Data; Development; Disease; Disruption; Employee Strikes; Focal Segmental Glomerulosclerosis; Foot Process; Functional disorder; Genes; Genetic; HIV-1; Human; Immunoglobulin Domain; Immunoglobulins; In Vitro; Infection; Integral Membrane Protein; Kidney; Kidney Diseases; Kidney Failure; Maps; Mediating; Microfilaments; Microtubules; Mouse Strains; Mus; Mutate; Names; Neurons; Orthologous Gene; Pathogenesis; Pathologic; Pattern; Phenotype; Play; Predisposition; Protein Overexpression; Proteins; Range; Renal glomerular disease; Research Personnel; Resistance; Role; Scaffolding Protein; Specificity; Synapses; Transgenes; Transgenic Mice; Transgenic Organisms; Up-Regulation; alpha Actinin; domain mapping; extracellular; glomerular basement membrane; in vivo; migration; mouse model; neuronal guidance; podocyte; prevent; programs; renal epithelium

DESCRIPTION (provided by applicant): HIV-associated nephropathy (HIVAN) is a leading cause of renal failure in Blacks. The main pathologic feature of HIVAN is collapsing glomerulopathy, which is caused by the direct infection of podocytes by HIV-1. Infected podocytes show unique phenotypic changes including dedifferentiation, increased proliferation, and changes in the actin cytoskeleton. We have cloned a host gene named sidekick-1 (sdk-1), a transmembrane protein of the immunoglobulin superfamily, as being highly upregulated in HIV-1 transgenic podocytes, and we believe sdk-1 to have a key role in HIVAN pathogenesis. We have shown that sdk-1 is highly upregulated in glomeruli in both HIV-1 transgenic mice and in human kidney biopsies. Our data also show that sdk-1 and its ortholog sidekick-2 (sdk-2) function as homophilic adhesion molecules in cells such that each sidekick prefers to bind to its own kind, and we have begun to map the domains responsible. Moreover, sdk-1 and sdk-2 were shown by others to function as neuronal guidance molecules, targeting axons to specific synapses; this suggests that sdk-1 and sdk-2 may analogously have a critical role in determining and maintaining glomerular architecture normally and in disease. In this proposal: 1. We will determine: a) the critical domain(s) of sdk-1 and sdk-2 responsible for the specificity of sdk-1 and sdk-2's homophilic interactions; b) whether targeted disruption of this critical domain's function in HIV-1 podocytes in vitro and in vivo can prevent HIV-1 induced phenotypic changes. 2. We will identify phenotypic changes in podocytes induced by sdk-1 expression. We will determine: a) to what extent does targeted overexpression of sdk-1 in podocytes in transgenic mice recapitulate HIVAN pathophysiology; b) how overexpression of sdk-1 in podocytes effects their ability to interact with glomerular basement membrane components; c) if sdk-1 upregulation induced by HIV-1 infection changes podocye-podocyte adhesion in vitro? 3. We will characterize the interaction of sdk-1 with the actin cytoskeleton. We will identify:a) the potential intracellular sdk-1 interacting proteins including alpha actinin-4; b) the effects of sdk-1 expression on organization of actin filaments and microtubules in podocytes and on expression of other actin-associated molecules. 4. We will better characterize expression patterns of sdk-1 in normal kidney, HIVAN and other glomerular diseases and examine differences in sdk-1 and sdk-2 expression between HIVAN susceptible and HIVAN resistant mouse strains. These studies should elucidate the mechanisms by which sdk-1 contributes to podocyte dysfunction in HIVAN.

描述（由申请人提供）： HIV相关性肾病（HIVAN）是黑人肾衰竭的主要原因。HIVAN的主要病理特征是由HIV-1直接感染足细胞引起的塌陷性肾小球病。感染足细胞表现出独特的表型变化，包括去分化，增殖增加，肌动蛋白细胞骨架的变化。我们已经克隆了一个名为sidekick-1（sdk-1）的宿主基因，它是免疫球蛋白超家族的一种跨膜蛋白，在HIV-1转基因足细胞中高度上调，我们相信sdk-1在HIVAN发病机制中具有关键作用。我们已经证明，sdk-1在HIV-1转基因小鼠和人肾活检的肾小球中高度上调。我们的数据还显示sdk-1和它的直系同源物sidekick-2（sdk-2）在细胞中起着嗜同性粘附分子的作用，因此每个sidekick都倾向于与自己的同类结合，我们已经开始绘制相关的结构域。此外，sdk-1和sdk-2被其他人证明作为神经元引导分子，将轴突靶向特定的突触;这表明sdk-1和sdk-2可能类似地在正常和疾病中决定和维持肾小球结构中起关键作用。在本提案中： 1.我们将确定：a）sdk-1和sdk-2的关键结构域负责sdk-1和sdk-2的亲同种相互作用的特异性; B）体外和体内靶向破坏HIV-1足细胞中该关键结构域的功能是否可以预防HIV-1诱导的表型变化。 2.我们将鉴定sdk-1表达诱导的足细胞表型变化。我们将确定：a）sdk-1在转基因小鼠足细胞中的靶向过表达在多大程度上再现了HIVAN病理生理学; B）sdk-1在足细胞中的过表达如何影响它们与肾小球基底膜组分相互作用的能力; c）HIV-1感染诱导的sdk-1上调是否改变了体外足细胞-足细胞粘附？ 3.我们将描述sdk-1与肌动蛋白细胞骨架的相互作用。我们将确定：a）潜在的细胞内sdk-1相互作用蛋白，包括α辅肌动蛋白-4; B）sdk-1表达对足细胞中肌动蛋白丝和微管组织的影响，以及对其他肌动蛋白相关分子表达的影响。 4.我们将更好地表征sdk-1在正常肾脏、HIVAN和其他肾小球疾病中的表达模式，并检查HIVAN易感和HIVAN抗性小鼠品系之间sdk-1和sdk-2表达的差异。 这些研究应该阐明sdk-1在HIVAN中促进足细胞功能障碍的机制。