The role of FAT10 in the pathogenesis of HIV-associated nephropathy

FAT10在HIV相关肾病发病机制中的作用

• 批准号: 7339445
• 负责人: MICHAEL J ROSS
• 金额: $ 34.23万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2012-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7339445
• 关键词: AIDS-Associated Nephropathy; African; African American; Alleles; Apoptosis; Biopsy Specimen; Breeding; Cells; Chronic Kidney Failure; Code; Development; Disease; Disease Progression; Epithelial; Epithelial Cells; Exons; FAT gene; Gene Deletion; Genes; Genetic; Genetic Polymorphism; HIV; HIV Infections; HIV-1; Haplotypes; Human; In Vitro; Individual; Inflammation; Kidney; Kidney Diseases; Kidney Failure; Knockout Mice; Laboratories; Lead; Mediator of activation protein; Mus; N-terminal; NF-kappa B; Pathogenesis; Patients; Phenotype; Phosphotransferases; Predisposition; Production; Proteins; Research Personnel; Risk; Role; Signal Transduction; Single Nucleotide Polymorphism; Stimulus; Testing; Transgenic Mice; Transgenic Model; Tubular formation; UBD protein; Ubiquitin Like Proteins; Up-Regulation; Variant; insight; knock-down; novel; prevent; programs; small hairpin RNA; vector

DESCRIPTION (provided by applicant): Tubulointerstitial renal disease is an important component of the pathobiology of HIV-associated nephropathy (HIVAN), the most common cause of chronic renal failure in HIV-infected individuals. In HIVAN, HIV infection of renal tubular epithelial cells (RTECs) leads to dysregulated apoptosis and production of proinflammatory molecules, both of which contribute importantly to progressive renal failure. HIVAN occurs almost exclusively in people of African ancestry and the genetic background of the individual is a critical determinant of whether renal epithelial infection by HIV will lead to progressive renal disease. We have recently demonstrated that the ubiquitin-like protein FAT10 is highly upregulated by HIV-infection of human RTECs in vitro and in HIVAN biopsy specimens, that FAT10 expression induces apoptosis in RTECs, and that preventing FAT10 expression with shRNA constructs ameliorates HIV-induced RTEC apoptosis. Preliminary studies in our laboratory have also demonstrated that FAT10 expression upregulates NF-KB signaling in human RTECs and may therefore induce production of proinflammatory mediators by these cells. We have identified four alleles of the FAT10 gene, one of which is significantly more common in patients with HIVAN. Since RTEC apoptosis and production of proinflammatory mediators by RTECs are important factors in HIVAN pathogenesis, we hypothesize that polymorphisms in the FAT10 gene alter its ability to induce RTEC apoptosis and NF-kB activation in RTECs. We will test these hypotheses in the following three Specific Aims: 1: To determine the effects of nonsynonymous FAT10 SNPs in FAT10 upon its subcellular localization and proapoptotic function. In this aim, we will determine how the four variants of the FAT10 protein differ in their subcellular localization and their ability to induce apoptosis. 2: To define the role of FAT10 in NF-KB signaling in human renal tubular epithelial cells and to determine if the FAT10 alleles differ in their ability to stimulate NF-KB signaling. In aim 2, we will study the mechanisms by FAT10 activates NF-KB signaling. 3: To determine whether FAT10 expression is necessary for the development of the HIVAN phenotype in the HIV-1 transgenic model of HIVAN. In aim 3, we will breed FAT10 knockout mice with HIV-1 transgenic mice and study the effect of FAT gene deletion upon the HIVAN phenotype. These studies will elucidate novel mechanisms of disease progression in HIVAN.

描述（由申请人提供）：肾小管间质肾脏疾病是HIV相关肾病（Hivan）病理生物学的重要组成部分，这是HIV感染者慢性肾衰竭的最常见原因。在Hivan中，肾小管上皮细胞（RTEC）的HIV感染导致凋亡失调和促炎分子的产生，这两种分子都对渐进性肾衰竭做出了重要贡献。 Hivan几乎完全出现在非洲血统的人群中，个人的遗传背景是HIV肾上皮感染是否会导致肾脏肾脏疾病的关键决定因素。我们最近证明，泛素样蛋白FAT10在体外和HIVAN活检标本中的HIV感染高度上调，即FAT10表达在RTEC中诱导凋亡，而SHRNA用SHRNA构建的FAT10表达可缓解HIV诱导的HIV诱导的RTEC RTEC凋亡。我们实验室的初步研究还表明，FAT10表达上调人RTEC中的NF-KB信号传导，因此可能诱导这些细胞促炎性介质的产生。我们已经确定了FAT10基因的四个等位基因，其中一个在Hivan患者中更为常见。由于RTEC的RTEC凋亡和RTEC促炎介质的产生是Hivan发病机理中的重要因素，因此我们假设FAT10基因中的多态性改变了其诱导RTEC凋亡和RTEC中NF-KB激活的能力。我们将在以下三个特定目的中检验这些假设：1：确定非同义FAT10 SNP在FAT10中对其亚细胞定位和促凋亡功能的影响。在此目标中，我们将确定脂肪10蛋白的四个变异在其亚细胞定位以及它们诱导凋亡的能力方面有何不同。 2：定义FAT10在NF-KB信号传导中的作用在人肾小管上皮细胞中，并确定FAT10等位基因刺激NF-KB信号传导的能力是否有所不同。在AIM 2中，我们将通过FAT10研究该机制会激活NF-KB信号传导。 3：确定在HIVAN的HIV-1转基因模型中，FAT10表达是否对于开发Hivan表型是必需的。在AIM 3中，我们将用HIV-1转基因小鼠繁殖Fat10敲除小鼠，并研究脂肪基因缺失对Hivan表型的影响。这些研究将阐明希文疾病进展的新机制。