VIRAL PATHOGENESIS OF HIV ASSOCIATED NEPHROPATHY

HIV 相关肾病的病毒发病机制

• 批准号: 6862320
• 负责人: Mary E. Klotman
• 金额: $ 31.8万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-07-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6862320
• 关键词: African American; HIV envelope protein; HIV infections; biopsy; caucasian American; clinical research; epithelium; genetic polymorphism; genetic susceptibility; genetically modified animals; human immunodeficiency virus 1; human subject; kidney disorder; laboratory mouse; laser capture microdissection; podocyte; protein sequence; protein structure function; racial /ethnic difference; renal failure; virus genetics; virus infection mechanism; virus protein

HIV-associated nephropathy (HIVAN) is now the third leading cause of renal failure in African Americans, the most common cause of chronic renal failure in HIV-1 infected individuals and renal disease is now the fourth leading cause of death in these patients. Accumulating evidence supports a direct role of HIV-1 infection of renal glomerular and tubule epithelial cells in HIVAN pathogenesis. Furthermore, quasispecies analysis of HIV-1 envelope sequences simultaneously derived from renal epithelial cells and peripheral blood mononuclear cells show renal-specific subclusters consistent with active replication of HIV-1 in the renal compartment. In vitro and transgenic mouse model data suggest that direct expression of HIV-1 genes, particularly nef, in renal epithelium can produce phenotypic changes and alterations in expression of cell genes involved in proliferation and differentiation that are consistent with changes associated with HIVAN. Project 2 will further define the direct role of infection of renal epithelial cells in HIVAN pathogenesis. To determine if renal epithelial infection alone can account for the disease, renal tissue will be examined from HIV-infected patients who have an alternative etiology of renal disease. This will include those of African descent as well as Caucasians. Examination of tissue will include in situ DNA PCR and confirmatory laser dissection of epithelial cells with PCR amplification of HIV sequences. To further our understanding of viral lentry into this unique compartment, we will phenotypically characterize the HIV-1 envelopes directly obtained from renal epithelium by laser capture dissection. In light of our previous published work and preliminary data demonstrating the effects of Nef and, to a lesser degree, Vpr on podocytes, we will use transgeneic mouse modeling to study the contribution of individual gene products expressed in specific cell types to pathogenesis. Targeted expression will be achieved either through direct expression of the transgene from site-specific promoters or the use of conditional transgenic constructs. Furthermore, nef sequences derived directly from renal epithelium will be genotypically and phenotypically characterized to determine if unique polymorphisms are associated with the development of HIVAN. The proposed studies should provide critical information regarding the role of epithelial infection in HIVAN pathogenesis and the interaction of HIV-1 with this unique reservoir and will impact on therapeutic interventions to prevent this devastating complication.

与艾滋病毒相关的肾病（Hivan）现在是非洲肾衰竭的第三大主要原因 美国人是HIV-1感染个体慢性肾衰竭和肾脏疾病的最常见原因，现在是这些患者的第四大死亡原因。积累的证据支持HIV-1感染HIVAN发病机理中肾肾小球和小管上皮细胞的直接作用。此外，同时源自肾上皮细胞和外周血单核细胞同时得出的HIV-1包膜序列的准分析显示，肾脏室中与HIV-1一致的肾脏特异性亚核细胞。体外和转基因小鼠模型的数据表明，肾上皮中HIV-1基因，特别是NEF的直接表达可以产生与与Hivan相关的变化一致的与增殖和分化有关的细胞基因表达的变化和变化。项目2将进一步定义肾上皮细胞在Hivan发病机理中感染的直接作用。为了确定肾上皮感染是否可以解释该疾病，将检查肾脏组织的肾脏感染患者，这些患者具有肾脏疾病的替代病因。这将包括非洲血统和高加索人的那些。组织的检查将包括原位DNA PCR和使用HIV序列PCR扩增的上皮细胞的确认激光解剖。为了进一步了解病毒式倾斜度，我们将表型从激光捕获的解剖中表征直接从肾上皮获得的HIV-1信封。鉴于我们以前发表的工作和初步数据，证明了NEF的影响，并且在较小程度上对Podocytes的影响，我们将使用经元小鼠建模来研究特定细胞类型对发病机理中表达的单个基因产物的贡献。有针对性的表达将通过从位点特异性启动子直接表达转基因或使用条件转基因构建体来实现。此外，直接从肾上皮得出的NEF序列将在基因型和表型上表征，以确定独特的多态性是否与Hivan的发展有关。拟议的研究应提供关键 有关上皮感染在Hivan发病机理中的作用以及HIV-1与这种独特储层的相互作用的信息，并将影响治疗干预措施以防止这种毁灭性的并发症。