The Role of Cholesterol in HIV Pathogenesis: Implications for NPC2

描述（由申请人提供）：先前的研究表明，HIV-1优先从富含胆固醇的微结构域（称为脂筏）萌发，并且胆固醇是HIV传染性和结构完整性所必需的。此外，胆固醇的存在也是HIV进入易感细胞和释放感染性病毒粒子所必需的。HIV感染病毒对胆固醇的需求使我们研究了HIV-1的萌芽和组装与胆固醇运输之间的关系。Niemann Pick Type C （NPC2）蛋白在细胞内胆固醇运输中起着至关重要的作用。鉴于病毒对胆固醇的需求和NPC2在胆固醇运输中的作用，我们假设NPC2的活性在HIV-1生物发生的晚期是必需的。HIV-1诱导的脂肪营养不良和随之而来的心血管综合征，以及胆固醇在病毒发病机制中的关键作用，使本研究与NIH消除HIV/AIDS健康差距和促进公众健康的使命高度相关。这个项目的发现也可能为其他与胆固醇相关的差异提供见解。为此，制定了以下具体目标：1)测试NPC2是否在HIV-1复制中起作用。2)以鼻咽癌为模型，探讨胆固醇转运影响病毒产生的机制。将用于实现这些目标的主要科学技术是Western分析，PCR，基因沉默，ELISA，流式细胞术和显微镜。

DESCRIPTION (provided by applicant): Previous studies have shown that HIV-1 preferentially buds from cholesterol-rich microdomains called lipid rafts, and that cholesterol is required for HIV infectivity and structural integrity. Moreover, the presence of cholesterol is also required both for HIV entry into susceptible cells as well as for release of infectious virions. The viral requirement of cholesterol for HIV infectivity led us to study the relationship between HIV-1 budding and assembly and cholesterol trafficking. Niemann Pick Type C (NPC2) protein plays a vital role in the intracellular trafficking of cholesterol. Given the viral requirement of cholesterol for productivity and given the role of NPC2 in cholesterol trafficking, we hypothesize that NPC2 activity is required in the late stages of HIV-1 biogenesis. HIV-1 induced lipodystrophy and the consequent cardiovascular syndromes, as well as the critical role of cholesterol in viral pathogenesis, make this study highly relevant to NIH's mission to eliminate HIV/AIDS health disparities and to promote public health. The findings from this project may also offer insight into other cholesterol- associated disparities. To this end, the following specific aims have been developed: 1) To test whether or not NPC2 plays a role in HIV-1 replication. 2) To examine the mechanism by which cholesterol trafficking effects viral production using NPC disease as a model. The primary scientific techniques that will be utilized to address these aims are Western analysis, PCR, gene silencing, ELISA, flow cytometry, and microscopy.