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

 描述(申请人提供)：这项研究利用疱疹病毒运输蛋白US9的自然特征作为一种新的方法来研究并可能调节与艾滋病毒相关的神经认知障碍(HAND)相关的脂筏依赖的细胞过程。由于脂筏代表了这些情况下可能涉及的关键分子事件的动态平台，我们的研究将利用US9的特性来追踪HIV诱导的脂筏变化，并改变淀粉样前体蛋白(APP)的加工-用于机械和干预目的。拟议的实验将检验这样的假设，即HIV-1gp120/Tat有利于淀粉样酶向膜微域的运输，导致神经毒性，并且这些病毒蛋白不影响US9的神经元内分布。这项工作将产生一种新的强大的策略，能够直接研究HIV-1蛋白诱导的脂筏变化和淀粉样蛋白形成之间的联系，以及可能发现一种减少神经元功能障碍的新的治疗方法。简而言之，我们将使用US9作为神经元蛋白质运输的分子示踪剂，以研究HIV-1gp120和Tat在参与神经毒性的脂筏相关蛋白质的神经元内分布中引起的变化，包括但不限于APP处理酶(目标1)。此外，通过US9介导的靶向改变APP加工的工程酶，我们将增加将APP加工转移到非淀粉样变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效应(目标2)。为了确定病毒蛋白在体外和体内的作用，这些目标包括在原代神经元培养和小动物模型中进行实验。如果成功，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，确定淀粉样蛋白发生和神经元存活/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，本研究的技术方法和研究范围都具有很高的创新性和重大意义，并有望对手部领域产生实质性的影响。

 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.