Tat endolysosome escape and HAND

项目摘要 我们对达特内溶酶体逃逸及其在肿瘤发病机制中的作用的理解存在根本性的差距。 HIV-1相关的神经认知障碍（HAND）。这里的目标是确定分子机制 达特通过何种方式逃逸内溶酶体，以及这种逃逸导致神经毒性的程度。我们将 测试我们的中心假设，即达特通过由V0扇区形成的蛋白孔逃逸内溶酶体。 v-ATP酶，这种逃逸是由内溶酶体双孔通道（TPC）释放的钙促进的， 这种钙依赖性达特内溶酶体逃逸过程，LTR的上游事件， 反式激活先于并促成Tat诱导的神经毒性。根据我们的初步调查结果， 新的假设将通过追求三个具体目标进行测试。(1)确定v-ATP酶的参与 达特内溶酶体逃逸中的TPC。(2)确定V-ATP酶和TPC在Tat诱导的细胞凋亡中的参与。 神经毒性(3)确定v-ATP酶和TPC在Tat诱导的学习和认知中的参与 损伤以及HAND样病理学。建议的工作具有很强的创新性 因为它关注于确定达特通过蛋白质孔逃离内溶酶体的机制 由v-ATP酶的V0区形成;这一过程可由内溶酶体释放的钙调节。 这种新的钙依赖性达特内溶酶体逃逸过程可能是一个早期和上游事件， Tat诱导的神经毒性。成功完成拟议的研究将为HIV-1提供新的见解 感染和HAND的发病机制，并可能产生新的和有效的治疗策略。

Project Abstract Fundamental gaps exist in our understanding in Tat endolysosome escape and its role in the pathogenesis of HIV-1 associated neurocognitive disorders (HAND). The objective here is to determine molecular mechanisms by which Tat escapes endolysosomes, and the extent to which this escape contributes to neurotoxicity. We will test our central hypothesis that Tat escapes endolysosomes through protein pores formed by the V0 sector of v-ATPase, that this escape is facilitated by calcium released from endolysosome two-pore channels (TPCs), and that such a calcium-dependent Tat endolysosome escape process, an upstream event of LTR transactivation, precedes and contributes to Tat-induced neurotoxicity. Guided by our preliminary findings, this novel hypothesis will be tested by pursuing three specific aims. (1) Determine the involvement of v-ATPase and TPCs in Tat endolysosome escape. (2) Determine the involvement of v-ATPase and TPCs in Tat-induced neurotoxicity. (3) Determine the involvement of v-ATPase and TPCs in Tat-induced learning and cognitive impairment as well as HAND-like pathology in Tat transgenic mice. The proposed work is highly innovative because it focuses on determining mechanisms by which Tat escapes endolysosomes via protein pores formed by V0 sector of v-ATPase; a process that can be regulated by calcium released from endolysosomes. Such a novel calcium-dependent Tat endolysosome escape process might be an early and upstream event of Tat-induced neurotoxicity. Successful completion of the proposed studies will provide novel insights into HIV-1 infection and the pathogenesis of HAND and may yield new and effective therapeutic strategies against HAND.