Exploring the Role of Semen Amyloids in Promoting HIV Infection and Fertilization

7.项目摘要/摘要： 艾滋病毒最常见的传播途径是性接触，而精液是全球 这种致命病毒的传播。我们的研究表明，精液非但不是艾滋病毒的被动传播媒介， 在体外显著增强艾滋病毒感染，我们进一步鉴定和表征了淀粉样纤维 来自人类精液，增加了HIV-1与其细胞靶点的融合。这些纤维可以增强HIV 感染超过几个数量级，因此是发展一种 局部使用的艾滋病毒杀微生物剂。这项提议的前两个目标集中在更好地描述这些机制的特征 通过这些精液纤维增强HIV的感染性及其对生殖器内存在的细胞的影响 粘膜。在目标1中，我建议使用神经生物学领域的技术来表征 最有效地增强艾滋病毒感染性的精液纤维的形态特征。此信息 将揭示精液中淀粉样蛋白构象的类型，在努力设计特定的 精液中抗宿主因子的抑制剂。在目标2中，我将确定精液纤维是否会导致 宿主细胞中的炎症，以及这在促进艾滋病毒感染方面可能起到的作用。众所周知， 炎症通常通过招募易感靶细胞和促进艾滋病毒传播来促进艾滋病毒的传播 基因转录。了解精液纤维在多大程度上促进宿主炎症 传播对于开发有效防止性传播的杀微生物剂至关重要 病毒。最后，提案的目标3侧重于更好地理解基本的生理学 精原纤维的功能。精液纤维没有进化到促进HIV感染，可能有生物学上的 人类的目标。耐人寻味的是，HIV与其细胞靶点的融合与 精子到卵子，增加了这些纤维在受精过程中发挥作用的可能性。在这个目标中，我 建议进行体外受精(IVF)和体内人工授精实验，以确定精液是否 我们已经确定的纤维可以促进小鼠配子的融合。了解这些纤维是否 促进受精是开发艾滋病毒杀微生物剂的重要信息，还可以 在生殖领域有重大影响。已经在以下方面投入了大量努力 开发一种有效的艾滋病毒杀微生物剂。然而，该领域仍然缺乏一种在以下方面高效的药物 防止艾滋病毒的性传播，部分原因是我们对分子事件缺乏基本了解 周围的粘膜传播艾滋病毒。这项建议侧重于更好地了解艾滋病毒的一个方面。 传播，即自然产生的精液纤维增强艾滋病毒感染性的效果。虽然 该提议仅限于对这些纤维的体外分析，我们正在启动恒河猴的平行实验。 以检查这些纤维在体内的效果。我在指导阶段的计划 如果K99/R00获得资金，它将开发淀粉样蛋白和生殖生物学方面的新技术，并 将这些技能应用于了解艾滋病毒的传播。这将通过努力实现 与我的K99导师和顾问委员会密切合作。我的长期目标是增进我们的理解 围绕着生殖器粘膜中艾滋病毒传播的分子事件，并将其翻译成 将知识转化为开发新类别的艾滋病毒预防药物。

7. PROJECT SUMMARY/ABSTRACT: HIV is most frequently transmitted following sexual contact, and semen is the vehicle fueling the global spread of this deadly virus. Far from being a passive vector for HIV, our research revealed that semen drastically enhances HIV infection in vitro, and we have further identified and characterized amyloid fibrils from human semen that increase HIV-1 fusion to its cellular targets. These fibrils can enhance HIV infection by over several orders of magnitude, and therefore serve as good targets for the development of a topical HIV microbicide. The first two aims of this proposal focus on better characterizing the mechanisms by which these semen fibrils enhance HIV infectivity and their influence on cells present within the genital mucosa. In Aim 1, I propose to use techniques from the field of neurobiology to characterize the morphological characteristics of semen fibrils that most effectively enhance HIV infectivity. This information will reveal the types of amyloid conformations in semen that should be targeted in efforts to design specific inhibitors against host factors in semen. In Aim 2, I will determine whether semen fibrils induce inflammation in host cells, and what role this may have in promoting HIV infection. It is known that inflammation generally facilitates HIV transmission by recruiting susceptible target cells and promoting HIV gene transcription. Understanding the extent to which semen fibrils contribute to host inflammation during transmission will be vital for developing microbicides that are effective in preventing sexual transmission of the virus. Lastly, Aim 3 of the proposal focuses on better understanding the fundamental physiological function of semen fibrils. Semen fibrils did not evolve to promote HIV infection, and may have a biological purpose in humans. Intriguingly, HIV fusion to its cellular target shares many properties with the fusion of a spermatozoon to an egg, raising the possibility for a role for these fibrils in fertilization. In this aim, I propose in vitro fertilization (IVF) and in vivo artificial insemination experiments to determine if the semen fibrils we have characterized promote the fusion of murine gametes. Understanding whether these fibrils serve to promote fertilization is vital information for the development of an HIV microbicide, and could also have a significant impact in the field of reproduction. Substantial efforts have been invested into developing an effective HIV microbicide. However, the field still lacks a drug that is highly effective at preventing the sexual spread of HIV, in part due to our lack of basic understanding of the molecular events surrounding mucosal HIV transmission. This proposal focuses on better understanding one aspect of HIV transmission, namely the effect of naturally-occurring semen fibrils that enhance HIV infectivity. Although the proposal is limited to in vitro analysis of these fibrils, we are initiating experiments in parallel in rhesus macaques to examine the effect of these fibrils in vivo. My plan during the mentored phase of the K99/R00, were it to get funded, is to develop new technical skills in amyloid and reproductive biology and to apply these skills towards understanding HIV transmission. This will be accomplished by working closely with my K99 mentors and advisory committee. My long-term goal is to advance our understanding of the molecular events surrounding HIV transmission in the genital mucosa, and to translate this knowledge into the development of new classes of HIV preventatives.