Transmigration of HIV-1-infected cells in NeuroAIDS

NeuroAIDS 中 HIV-1 感染细胞的迁移

• 批准号: 6893204
• 负责人: Kevin Jon Williams
• 金额: $ 23.4万
• 依托单位: THOMAS JEFFERSON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2004
• 资助国家: 美国
• 起止时间: 2004-12-20 至 2006-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6893204
• 关键词: AIDS; AIDS dementia complex; RNA interference; T lymphocyte; antihypercholesterolemic agent; blood brain barrier; cell migration; chemokine; chemoprevention; cytokine; gene expression; genetic regulation; helper T lymphocyte; human immunodeficiency virus 1; human subject; matrigel; membrane model; metalloendopeptidases; microarray technology; monocyte; neuropathology; protein biosynthesis; proteomics; tissue /cell preparation; transport inhibitor; virus infection mechanism

DESCRIPTION (provided by applicant): Human immunodeficiency virus type 1 (HIV-1) enters the brain soon after serconversion. Consequently, the central nervous system (CNS) becomes a sanctuary for virus, and it can be damaged by virus or virally encoded proteins. A prominent model for viral entry into the CNS is the "Trojan Horse" hypothesis, in which HTV- 1-infected CD4+ T-lymphocytes and monocytes transmigrate across the blood-brain barrier (BBB). Consistent with prior work, we observed that HIV-1 infection enhances T-cell and monocyte transmigration in vitro across a simple acellular barrier of Matrigel, which mimics basement membrane. More importantly, we made the novel discovery that clinically available inhibitors of cholesterol biosynthesis (statins) potently inhibit HTV-1 -induced transmigration. This finding arose from an essential synergy between two laboratories in different fields, lipid metabolism (Dr. Williams) and neurovirology (Drs. Mukhtar and Pomerantz). In addition, as noted in the Introduction and revised Preliminary Studies, we recently found a neuroprotective effect of statins as well. Our central hypothesis is that HIV-1 infection alters the expression of specific genes that play a central role in transmigration and cytotoxicity, and that statins restore the expression of these genes towards normal levels. Experimental systems available to us include primary human T-cells and monocytes, our acellular model barrier of Matrigel, and a sophisticated cellular BBB model that we created using well-characterized human BMVECs and astrocytes grown in transwell inserts over human neurons. There are two Specific Aims: Aim I; Molecular mechanisms by which statins reduce transmigration of HIV-1-infected CD4+ T-cells and monocytes through an acellular model basement membrane. Three sub-Aims will study whether this effect results from (i) cholesterol-dependent or -independent (pleiotropic) effects of statins, (ii) involvement of specific genes identified on our focused gene array as altered by infection but restored by statins, e.g., MMPs, TIMPs, paxillin, and rho-related proteins, and (iii) alterations in specific inflammatory or cytotoxic cytokines. Aim II: Effects of statins on transmigration of HIV-1-infected T-cells and monocytes through a cellular blood brain barrier model in vitro. Sub-aims will somewhat parallel Aim I, focusing on the effects of infection and statin treatment on specific gene expression, cytokine release, endothelial integrity, and transmigration. Overall, our proposed studies will provide new insights into HIV-1 neuropathogenesis. In addition, our work may provide a new use for statins to prevent or treat AIDS-related dementia and to deplete or eradicate the CNS as an HIV-1 sanctuary.

描述（由申请人提供）：人类免疫缺陷病毒1型（HIV-1）在血清转化后不久进入大脑。因此，中枢神经系统（CNS）成为病毒的避难所，它可以被病毒或病毒编码的蛋白质破坏。病毒进入CNS的一个突出模型是“特洛伊木马”假说，其中HIV- 1感染的CD 4 + T淋巴细胞和单核细胞穿越血脑屏障（BBB）。与先前的工作一致，我们观察到HIV-1感染增强了T细胞和单核细胞在体外穿过一个简单的无细胞基质胶屏障的迁移，该屏障模拟基底膜。更重要的是，我们有了新的发现，临床上可用的胆固醇生物合成抑制剂（他汀类药物）有效地抑制HIV-1诱导的迁移。这一发现源于两个实验室在不同领域的重要协同作用，脂质代谢（威廉姆斯博士）和神经病毒学（Mukhtar和Pomerantz博士）。此外，正如引言和修订的初步研究中所指出的，我们最近发现他汀类药物也具有神经保护作用。我们的中心假设是，HIV-1感染改变了特定基因的表达，这些基因在细胞迁移和细胞毒性中起着核心作用，他汀类药物使这些基因的表达恢复到正常水平。我们可用的实验系统包括原代人T细胞和单核细胞，我们的Matrigel非细胞模型屏障，以及我们使用在transwell插入物中生长在人神经元上的良好表征的人BMVEC和星形胶质细胞创建的复杂细胞BBB模型。有两个具体目标：目标一;他汀类药物减少HIV-1感染的CD 4 + T细胞和单核细胞通过非细胞模型基底膜迁移的分子机制三个子目标将研究这种效应是否由以下因素引起：（i）他汀类药物的胆固醇依赖性或非依赖性（多效性）效应，（ii）在我们的聚焦基因阵列上鉴定为被感染改变但被他汀类药物恢复的特定基因的参与，例如，MMP、TIMP、桩蛋白和rho相关蛋白，以及（iii）特异性炎症或细胞毒性细胞因子的改变。目的II：他汀类药物对HIV-1感染的T细胞和单核细胞通过细胞血脑屏障模型的影响。子目标将在某种程度上平行于目标I，重点关注感染和他汀类药物治疗对特定基因表达、细胞因子释放、内皮完整性和迁移的影响。总的来说，我们提出的研究将为HIV-1神经发病机制提供新的见解。此外，我们的工作可能为他汀类药物提供新的用途，以预防或治疗艾滋病相关的痴呆症，并消耗或根除CNS作为HIV-1避难所。