Epithelial gene expression in HIV-associated nephropathy

HIV相关肾病的上皮基因表达

• 批准号: 6669123
• 负责人: MICHAEL J ROSS
• 金额: $ 12.78万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2002
• 资助国家: 美国
• 起止时间: 2002-09-30 至 2007-09-29
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6669123
• 关键词: HIV infections; clinical research; epithelium; gene expression; genetic mapping; genetically modified animals; host organism interaction; human immunodeficiency virus 1; human tissue; kidney disorder; laboratory mouse; microarray technology; recombinant virus; renal tubule; representational difference analysis

DESCRIPTION (provided by applicant): HIV-associated nephropathy (HIVAN) is the most common cause of chronic renal failure in HIV-1 infected patients. The most prominent histologic feature of HIVAN is dilatation of the renal tubules. Investigators in our laboratory have previously demonstrated, in biopsy specimens from patients with HIVAN, that renal tubular epithelial cells are infected with HIV-1. However, the cellular events following HIV-1 infection, contributing to progressive renal failure in HIVAN are not well understood. We have demonstrated previously, that HIV-1 infection may involve epithelial cells from several portions of the nephron, including the proximal tubule. We have isolated proximal tubular epithelial cells from HIVAN biopsy specimens and conditionally immortalized them with the temperature-sensitive SV40 large T antigen. These cells (HPT-1) grow indefinitely under permissive conditions (33C), and when differentiated at 37C, express typical proximal tubular phenotypic markers. The purpose of the proposed-studies is to determine the host genes that are differentially expressed following infection of renal tubular epithelial cells with HIV-1. Once candidate host genes are identified, we will map the HIV-1 responsible for inducing altered expression of these genes. In the proposed studies, we will use a VSV-pseudotyped replication defective HIV-1 gag/pol deletion construct to transduce HPT-1 cells. Following transduction, we will use representational difference analysis (RDA) and oligonucleotide expression micrarrays to determine the cellular genes that are differentially expressed following transduction by HIV-1. Differential expression of cellular genes will be confirmed by standard molecular techniques. Once host candidate genes are identified, we will transduce HPT-1 cells with a series of HIV-1 multigenic mutant constructs to map the specific HIV-1 genes responsible for the observed alterations in cellular gene expression. These studies should elucidate essential mechanisms of HIV-1 pathogenesis in HIVAN. It is also possible that genes implicated in the pathogenesis of HIVAN maybe important factors contributing to the marked predisposition of African-Americans to nearly all causes of renal failure.

描述（由申请人提供）： 与HIV相关的肾病（Hivan）是HIV-1感染患者慢性肾衰竭的最常见原因。 Hivan最突出的组织学特征是肾小管的扩张。我们实验室中的研究人员以前已经证明，在Hivan患者的活检标本中，肾小管上皮细胞被HIV-1感染。然而，HIV-1感染后的细胞事件，导致Hivan进行性肾衰竭的导致尚未得到充分了解。 我们先前已经证明，HIV-1感染可能涉及来自肾单位几个部分的上皮细胞，包括近端小管。我们已经从HIVAN活检标本中分离出近端管状上皮细胞，并用温度敏感的SV40大T抗原使它们有条件永生。这些细胞（HPT-1）在允许的条件下无限期生长（33C），当在37C时区分时，会表达典型的近端管状表型标记。 所提出的研究的目的是确定肾小管上皮细胞用HIV-1差异表达的宿主基因。一旦确定了候选宿主基因，我们将绘制负责诱导这些基因表达改变的HIV-1。 在拟议的研究中，我们将使用VSV型型复制有缺陷的HIV-1 GAG/POL缺失构建体来传递HPT-1细胞。转导后，我们将使用表示差异分析（RDA）和寡核苷酸表达微箭头来确定HIV-1转导后差异表达的细胞基因。细胞基因的差异表达将通过标准分子技术证实。一旦鉴定出宿主候选基因，我们将用一系列HIV-1多基因突变体构建体将HPT-1细胞转导HPT-1细胞，以绘制负责观察到的细胞基因表达改变的特定HIV-1基因。 这些研究应阐明Hivan中HIV-1发病机理的基本机制。还可能与Hivan的发病机理有关的基因可能导致非洲裔美国人明显倾向于几乎所有肾衰竭原因。