Transmigration of HIV-1-infected cells in NeuroAIDS

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。病毒进入中枢神经系统的一个重要模型是“特洛伊木马”假说，其中HTV- 1感染的CD4+ t淋巴细胞和单核细胞通过血脑屏障（BBB）转运。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简单的脱细胞屏障Matrigel，它模仿基底膜。更重要的是，我们发现临床上可用的胆固醇生物合成抑制剂（他汀类药物）能有效抑制HTV-1诱导的转运。这一发现源于两个不同领域的实验室之间的重要协同作用，脂质代谢（Williams博士）和神经病毒学（Williams博士）。Mukhtar and Pomerantz)。此外，正如引言和修订的初步研究中所指出的，我们最近也发现了他汀类药物的神经保护作用。我们的中心假设是HIV-1感染改变了在细胞迁移和细胞毒性中起核心作用的特定基因的表达，而他汀类药物使这些基因的表达恢复到正常水平。我们现有的实验系统包括原代人t细胞和单核细胞，我们的Matrigel脱细胞屏障模型，以及我们使用在人类神经元上的transwell插入物中生长的具有良好特征的人类bmvec和星形胶质细胞创建的复杂细胞血脑屏障模型。有两个具体目标：第一目标；他汀类药物通过脱细胞模型基底膜减少hiv -1感染的CD4+ t细胞和单核细胞转移的分子机制三个子目标将研究这种效应是否来自(i)他汀类药物的胆固醇依赖性或非依赖性（多效性）效应，（ii）在我们的重点基因阵列上发现的特定基因的参与，这些基因被感染改变，但被他汀类药物恢复，例如MMPs， TIMPs， paxillin和rho相关蛋白，以及（iii）特定炎症或细胞毒性细胞因子的改变。目的二：他汀类药物通过体外细胞血脑屏障模型对hiv -1感染的t细胞和单核细胞转移的影响。子目标将在一定程度上与目标1相似，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、内皮完整性和转运的影响。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预防或治疗艾滋病相关痴呆以及耗尽或根除作为HIV-1避难所的中枢神经系统提供新的用途。

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.