Effects of HIV-1 neurotoxins on lipid rafts-associated proteins

 描述（由申请人提供）：本研究利用疱疹病毒转运蛋白US 9的天然特征作为研究并可能调节与HIV相关神经认知障碍（HAND）相关的脂筏依赖性细胞过程的新方法。由于脂筏代表了可能参与这些疾病的关键分子事件的动态平台，我们的研究将利用US 9特性来追踪HIV诱导的脂筏变化，并改变淀粉样前体蛋白（APP）加工-用于机械和干预目的。拟议的实验将测试的假设，即HIV-1 gp 120/达特有利于运输淀粉样蛋白酶的膜微区，导致神经毒性，这些病毒蛋白不影响神经元内分布的US 9。这项工作将产生一个新的和强大的策略，能够直接研究HIV-1蛋白诱导的脂筏变化和淀粉样蛋白生成之间的联系，以及可能发现一种新的治疗方法，以减少神经元功能障碍。简而言之，我们将使用US 9作为神经元蛋白转运的分子示踪剂，以研究由HIV-1 gp 120和达特诱导的参与神经毒性的脂筏相关蛋白的神经元内分布的改变-包括但不限于APP加工酶（Aim 1）。此外，通过US 9介导的改变APP加工的工程酶的靶向，我们将增加将APP加工转向非淀粉样蛋白生成途径的细胞酶的表达，并测试HIV蛋白在这些条件下的神经毒性作用（目的2）。为了确定病毒蛋白的体外和体内作用，这些目标包括在原代神经元培养物和小动物模型中的实验。如果成功的话，这项研究将有助于描述脂筏变化在HIV诱导的神经元损伤中的作用，确定淀粉样蛋白生成和神经元存活/损伤之间的联系，并导致潜在的应用，旨在减少疾病晚期有害蛋白的局部积累。因此，无论是技术方法和范围，这项研究是高度创新和重大的，并预计将产生重大影响的领域的手。

 DESCRIPTION (provided by applicant): This research exploits natural features of the herpesvirus transport protein US9 as a novel approach to study, and possibly modulate, lipid rafts dependent cellular processes relevant to HIV Associated Neurocognitive Disorders (HAND). As lipid rafts represent dynamic platforms for key molecular events likely involved in these conditions, our studies will capitalize on US9 properties to trace HIV-induced changes in lipid rafts, and alter Amyloid Precursor Protein (APP) processing - for both mechanistic and interventional purposes. The proposed experiments will test the hypothesis that HIV-1 gp120/tat favor transport of amyloidogenic enzymes to membrane microdomains, leading to neurotoxicity, and that these viral proteins do not affect intraneuronal distribution of US9. This work will generate a new and powerful strategy capable of studying directly the link between HIV-1 proteins induced changes of lipid rafts and amyloidogenesis, as well as possibly discovering a novel therapeutic approach to decreasing neuronal dysfunction. Briefly, we will use US9 as a molecular tracer of neuronal protein transport to study alterations induced by HIV-1 gp120 and tat in the intraneuronal distribution of lipid rafts-associated proteins that are involved in neurotoxiciy - including, but not limited to, APP processing enzymes (Aim 1). Moreover, through US9-mediated targeting of engineered enzymes altering APP processing, we will increase expression of cellular enzymes that shift APP processing toward the non-amyloidogenic pathway and test the neurotoxic effects of the HIV proteins under these conditions (Aim 2). In order to determine both in vitro and in vivo effects of the viral proteins, these aims include experiments in primary neuronal cultures and small animal models. If successful, this research will help delineate the role of lipid rafts changes in HIV-induced neuronal damage, determine the link between amyloidogenesis and neuronal survival/injury, and lead to potential applications intended to reduce local accumulation of noxious proteins at advanced stages of disease. Thus, both the technical approach and scope of this research are highly innovative and significant, and expected to exert a substantial impact on the field of HAND.