PROTECTING CNS CELLS FROM HIV AND HIV-INDUCED INJURY

描述（由申请人提供）：在这个R21资助申请中，我们在这里提议研究基因传递抑制HIV复制和HIV对CNS细胞的毒性。艾滋病毒脑病是艾滋病的一种常见且具有潜在破坏性的并发症。HW脑病的发展在很大程度上反映了HIV感染的脑细胞，特别是单核细胞来源的巨噬细胞（MDM）和小胶质细胞。它也可能涉及某些HIV蛋白对神经元的毒性。我们将使用基因传递技术来研究在原代培养的人MDM和小胶质细胞中抑制H1V感染和复制，并检查神经元免受HIV gp120包膜糖蛋白诱导凋亡的能力的保护。从talphag缺失的SV40 （rSV40s）衍生的重组基因递送载体永久转导HW未选择的关键中枢神经系统靶点，包括小胶质细胞、神经元和单核细胞源性巨噬细胞（MDM），体外效率为>98%。我们发现HIV在MDM和小胶质细胞中复制，携带HIV抑制转基因的rSV40s保护这些细胞免受HIV复制。我们还发现gp120在培养的神经元中引起细胞凋亡，而下调细胞膜CXCR4或提供过氧化氢酶等抗氧化酶的rSV40载体可以保护这些神经元。我们建议利用这些有希望的发现来研究rSV40基因传递以保护CNS细胞免受HIV感染。为了验证这一假设，我们提出以下具体目标：1.)确定在中枢神经系统细胞中单独或联合抑制HIV-1的最佳转基因。2)利用基因传递保护nt2来源的神经元免受HIV基因产物诱导的凋亡。尽管中枢神经系统艾滋病毒感染的频率和可怕的后果，患有艾滋病毒脑病的人几乎没有治疗选择。我们建议使用rSV40基因递送到中枢神经系统来解决这一治疗挑战，既保护大脑免受HIV感染，又减轻HIV诱导的中枢神经系统功能障碍。

DESCRIPTION (provided by applicant): In this R21 grant application, we propose here to study gene delivery to inhibit HIV replication and HIV toxicity for CNS cells. HIV encephalopathy is a common and potentially devastating complication of AIDS. Development of HW encephalopathy largely reflects HIV infection of brain cells, especially monocyte-derived macrophages (MDM) and microglia. It also may involve toxicity of certain HIV proteins for neurons. We will use gene delivery techniques to study inhibition of H1V infection and replication in primary cultured human MDM and microglia, and to examine protection of neurons from the ability of HIV gp120 envelope glycoprotein to induce apoptosis. Recombinant gene delivery vehicles derived from Talphag-deleted SV40 (rSV40s) permanently transduce unselected key CNS targets for HW, including microglia, neurons, and monocyte-derived macrophages (MDM) with >98% efficiency in vitro. We have found that HIV replicates in MDM and microglia, and that rSV40s carrying HIV inhibitory transgenes protect these cells from HIV replication. We have found as well that gp120 causes apoptosis in cultured neurons, and that rSV40 vectors that downregulate cell membrane CXCR4 or that provide antioxidant enzymes, such as catalase, protect these neurons. We propose to exploit these promising findings to study rSV40 gene delivery to protect the CNS cells from HIV. This proposal is based on the following hypothesis: To test this hypothesis, we propose the following specific aims: 1.) Identify optimal transgenes individually and in combination to inhibit HIV-1 in CNS cells. 2.) use gene delivery to protect NT2-derived neurons from apoptosis induced by HIV gene products. Despite the frequency and dire consequences of CNS HIV infection, people afflicted with HIV encephalopathy have few treatment options. We propose to address this therapeutic challenge using rSV40 gene delivery to the CNS, both to protect the brain from HIV infection and to mitigate HIV-induced CNS dysfunction.