The effect of HIV and cocaine abuse on semen exosome composition and function

摘要 艾滋病流行病仍然是一个重大的全球问题，目前有3 330多万人感染艾滋病。 感染艾滋病毒和艾滋病毒相关疾病的人每年夺去180多万人的生命。 各种研究表明，滥用包括可卡因在内的非法药物与吸毒人数增加之间存在联系。 艾滋病毒传播的风险，加速艾滋病毒疾病的进展和艾滋病相关的死亡率。妇女 是新增艾滋病毒病例增长最快的人群。据估计，超过80%的艾滋病毒 性传播导致病例，精液是主要的传播媒介。但只有0.1%的 与受感染个体的异性性交行为导致HIV感染，这表明 精液中所含的蛋白质可以调节精液中HIV颗粒的感染性或适合性。底层 艾滋病毒的传播机制还不清楚。尽管艾滋病研究取得了进展 发病机理和各种药剂抑制病毒感染的能力，预防HIV的方法 传播仍然需要更多的调查。最近，我们发现健康捐献者的精液 含有在大小方面形态上不同的外来体的异源群体 和电子密度。这些外来体含有不同种类的RNA（mRNA和小RNA）， 抑制HIV感染。我们的初步数据显示，类似于来自精液的外泌体， 健康供体，从HIV感染供体纯化的外泌体有效地阻断HIV感染， 来自使用可卡因的HIV感染供体的外来体不抑制HIV。我们的初步研究进一步 HIV感染和可卡因使用调节精液外泌体的RNA组成。 重要的是，来自HIV阴性和HIV感染供体的裂解精液外泌体，而不是供体 使用可卡因的人在无细胞系统中抑制裂解的HIV的逆转录，阻断 HIV U 5 DNA和U 5到Gag的逆转录。这些数据表明精液的直接影响 外泌体对HIV逆转录的影响。在本申请中，研究旨在1）确定 HIV感染和可卡因滥用对精液外泌体抗HIV活性的影响，2）确定 精液外泌体抗HIV作用的机制; 3）描述供体HIV的作用 状态和可卡因使用对精液外泌体组成影响。由于性传播是艾滋病的主要传播途径， 艾滋病毒和精液的传播是主要的载体，本申请中提出的研究将 解决了评估精液外泌体作为保护因子的功效的新挑战 以及捐赠者的艾滋病毒状况和可卡因使用如何调节这一功能。

Abstract The AIDS pandemic remains a significant global problem with over 33.3 million persons currently infected with HIV and HIV-related illnesses claiming the lives of over 1.8 million individuals annually. Various studies have suggested a link between abuse of illicit drugs, including cocaine and increased risk of HIV transmission, accelerated HIV disease progression, and AIDS-related mortality. Women represent the fastest growing population of new HIV cases. It is estimated that more than 80% of HIV cases result from sexual transmission and semen is the primary vector. However, only about 0.1% of acts of heterosexual coitus with an infected individual result in HIV infection, suggesting that factors contained in semen may modulate the infectivity or fitness of HIV particles in semen. The underlying mechanisms of HIV transmission are not well understood. In spite of the progress in studies of HIV pathogenesis and the ability of various agents to suppress virus infection, methods to prevent HIV transmission still require more investigation. Recently, we showed that semen of healthy donors contain heterologous populations of exosomes that are morphologically distinct with respect to size and electron density. These exosomes contain different species of RNA (mRNA and small RNA), and inhibit infection with HIV. Our preliminary data reveal that similar to exosomes from the semen of healthy donors, exosomes purified from HIV infected donors potently block HIV infection while exosomes from HIV infected donors who use cocaine do not inhibit HIV. Our preliminary study further showed that HIV infection and cocaine use modulates RNA composition of semen exosomes. Importantly, lysed semen exosomes from both HIV negative and HIV infected donors but not donors who used cocaine inhibit reverse transcription of lysed HIV in a cell-free system, block the synthesis of HIV U5 DNA, and reverse transcription of U5 to Gag. These data indicate direct effect of semen exosomes on HIV reverse transcription. In this application, studies are designed to 1) determine the effect of HIV infection and cocaine abuse on the anti-HIV activity of semen exosomes, 2) determine the mechanisms of anti-HIV effect of semen exosomes, and 3) characterize the effects of donor HIV status and cocaine use on semen exosome composition. As sexual transmission is the main route for the spread of HIV and semen is the primary vector, the studies proposed in this application will address a new challenge for evaluation of the efficacy of semen exosomes as a protective factor against HIV and how donor HIV status and cocaine use modulate this function.