Co-evolution of HIV-1 Regulators in CNS Disease

HIV-1 调节因子在中枢神经系统疾病中的共同进化

• 批准号: 6884008
• 负责人: Brian Wigdahl
• 金额: $ 35.63万
• 依托单位: DREXEL UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-04-15 至 2009-03-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6884008
• 关键词: AIDS /HIV neuropathy; CD4 molecule; biochemical evolution; cell line; gene expression; genetic registry /resource /referral center; genetic regulatory element; genotype; human fetus tissue; human immunodeficiency virus 1; immunocytochemistry; molecular biology information system; molecular cloning; nucleic acid repetitive sequence; polymerase chain reaction; protein binding; protein sequence; protein structure function; virus DNA; virus genetics; virus infection mechanism; virus protein; virus replication

DESCRIPTION (provided by applicant): HIV-1-associated pathogenesis in the immune system and central nervous system (CNS) is dependent on viral replication, which can be controlled by viral binding and entry, receptor/co-receptor usage, host cell proliferation and activation, cellular differentiation, antiretroviral therapy status, and regulation of viral expression by the long terminal repeat (LTR) in conjunction with viral and cellular regulatory proteins. Viral proteins that regulate LTR function and viral expression include the Tat trans-activator protein and viral protein R (Vpr), which can induce LTR activity through several cis-acting LTR elements. The replicative capacity of HIV-1 is further affected by evolution of the viral genotype (including tat, vpr, and the LTR) during the course of infection. The hypothesis of these investigations is that co-evolved viral regulatory elements (CVRE) Tat, Vpr, and the LTR alter the viral replicative phenotype, affect the spread of virus between compartmentalized cell types, and affect the course and severity of CNS disease. The specific aims of this application are to (1) establish a CVRE sequence database and a bank of DNA clones derived from HIV-1-infected immune cells (activated T cells, non-activated T cells, monocytic cells) and CNS-resident cells (microglia, perivascular macrophages, astrocytes) isolated from patients that differ in HAART and dementia status; (2) analyze the CVRE database for intrapatient variations (specific to cell type or compartmentalized to the CNS or immune system) and interpatient variations (HAART and dementia status-specific) in co-evolved Vpr, Tat, and LTR sequences; (3) construct infectious molecular clones (IMCs) incorporating select CVRE sequences and analyze each IMC-derived virus for CD4 binding, co-receptor utilization, and replication phenotype in immune- and CNS-derived cell lines and primary cells; and (4) determine the ability of co-evolved Tat-LTR, Vpr-LTR, and Tat-Vpr-LTR combinations from brain-derived CVRE clones to support transient expression, and establish correlations between transient LTR induction, dementia status, and HAART status. This unique approach to studying regulated HIV-1 expression in the context of CNS disease and HAART will provide greater insights into HIV-1-associated pathogenesis and may lead to more efficacious treatments of HIV-1 CNS disease.

描述(申请人提供)：免疫系统和中枢神经系统(CNS)中与HIV-1相关的发病机制依赖于病毒复制，病毒复制可通过病毒结合和进入、受体/辅助受体的使用、宿主细胞的增殖和激活、细胞分化、抗逆转录病毒治疗状态以及通过结合病毒和细胞调节蛋白的长末端重复序列(LTR)调节病毒表达来控制。调节LTR功能和病毒表达的病毒蛋白包括TAT反式激活蛋白和病毒蛋白R(VPR)，它们可以通过几个顺式作用的LTR元件诱导LTR活性。HIV-1的复制能力进一步受到感染过程中病毒基因(包括TAT、VPR和LTR)进化的影响。这些研究的假设是，共同进化的病毒调节元件(CVRE)TAT、VPR和LTR改变了病毒的复制表型，影响了病毒在不同细胞类型之间的传播，并影响了CNS疾病的进程和严重程度。这项应用的具体目的是(1)建立来自HIV-1感染的免疫细胞(激活的T细胞、非激活的T细胞、单核细胞)和从HAART和痴呆状态不同的患者分离的CNS驻留细胞(小胶质细胞、血管周围巨噬细胞、星形胶质细胞)的CVRE序列数据库和DNA克隆库；(2)分析CVRE数据库中共同进化的VPR、TAT和LTR序列中的患者内变异(特定于细胞类型或划分为CNS或免疫系统)和患者间变异(HAART和痴呆状态特定)；(3)构建含有精选CVRE序列的感染性分子克隆(IMCs)，分析每个IMC衍生病毒在免疫和中枢神经系统衍生细胞系和原代细胞中的CD4结合、共受体利用和复制表型；以及(4)从脑源性CVRE克隆中确定共同进化的TAT-LTR、VPR-LTR和TAT-VPR-LTR组合支持瞬时表达的能力，并建立瞬时LTR诱导、痴呆状态和HAART状态之间的关联。这种在CNS疾病和HAART背景下研究HIV-1调控表达的独特方法将提供对HIV-1相关发病机制的更多洞察，并可能导致对HIV-1 CNS疾病的更有效治疗。