Transmigration of HIV-1-infected cells in NeuroAIDS

描述(申请人提供)：人类免疫缺陷病毒1型(HIV-1)在血清转换后不久进入大脑。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突出模型是“特洛伊木马”假说，在该假说中，HTV-1感染的CD4T淋巴细胞和单核细胞跨越血脑屏障(BBB)。与之前的工作一致，我们观察到HIV-1感染在体外促进T细胞和单核细胞通过简单的Matrigel无细胞屏障迁移，Matrigel模仿基底膜。更重要的是，我们取得了新的发现，临床上可用的胆固醇生物合成抑制剂(他汀类)有效地抑制HTV-1诱导的移位。这一发现源于脂代谢(威廉姆斯博士)和神经病毒学(穆赫塔尔和波美兰茨博士)这两个不同领域的实验室之间的重要协同作用。此外，正如在导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感染改变了在转生和细胞毒性中发挥核心作用的特定基因的表达，他汀类药物将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类T细胞和单核细胞，我们的无细胞Matrigel模型屏障，以及我们使用具有良好特性的人BMVEC和生长在Transwell插入物中的星形胶质细胞在人类神经元上生长的复杂的细胞BBB模型。有两个特定的目标：目的I；他汀类药物通过无细胞模型基底膜减少HIV-1感染的CD4T细胞和单核细胞迁移的分子机制。三个子目标将研究这种效应是否源于(I)他汀类药物的胆固醇依赖或非依赖性(多效性)作用，(Ii)在我们的重点基因阵列上发现的被感染改变但被他汀类药物修复的特定基因的参与，例如MMPs、TIMPs、paxlin和Rho相关蛋白，以及(Iii)特定的炎症或细胞毒性细胞因子的改变。目的II：通过体外细胞血脑屏障模型观察他汀类药物对HIV-1感染的T细胞和单核细胞迁移的影响。子目标将在某种程度上与目标I类似，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、内皮完整性和移行的影响。总体而言，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性痴呆，并耗尽或根除中枢神经系统作为HIV-1避难所的作用。

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.