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

7.项目总结/摘要： 艾滋病毒最常在性接触后传播，精液是全球艾滋病毒传播的媒介。 这种致命病毒的传播。我们的研究表明，精液远不是艾滋病病毒的被动载体， 在体外实验中，它能显著增强HIV感染，我们进一步鉴定和表征了淀粉样纤维 可以增加HIV-1与细胞靶点的融合这些纤维可以增强艾滋病毒 感染超过几个数量级，因此可以作为发展的良好目标。 外用艾滋病毒杀微生物剂本提案的前两个目标侧重于更好地描述机制的特征 这些精液原纤维增强了艾滋病毒的感染力，并对生殖器内的细胞产生了影响， 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 精液原纤维的形态特征，最有效地提高艾滋病毒的感染性。这些信息 将揭示精液中淀粉样蛋白构象的类型，这应该是设计特异性 精液中的宿主因子抑制剂。在目标2中，我将确定精液原纤维是否诱导 宿主细胞中的炎症，以及这在促进HIV感染中可能起什么作用。已知的是 炎症通常通过招募易感靶细胞和促进HIV传播来促进HIV传播。 基因转录了解精液原纤维在多大程度上有助于宿主炎症， 传播将是至关重要的开发杀微生物剂，有效地防止性传播 病毒最后，该提案的目标3侧重于更好地理解基本的生理 精液纤维的功能精液原纤维没有进化到促进艾滋病毒感染，可能有生物学上的影响。 人类的目的。有趣的是，HIV与其细胞靶点的融合与HIV与其细胞靶点的融合具有许多特性。 精子到卵子，提高了这些纤维在受精中的作用的可能性。为此，我 提出体外受精（IVF）和体内人工授精实验，以确定精液是否 我们已经表征的原纤维促进鼠配子的融合。了解这些纤维 有助于促进受精是开发HIV杀微生物剂的重要信息， 在生殖领域产生了重大影响。投入了大量的努力， 开发一种有效的艾滋病毒杀菌剂。然而，该领域仍然缺乏一种高效的药物， 防止艾滋病毒的性传播，部分原因是我们缺乏对分子事件的基本了解 粘膜艾滋病毒传播。这项建议的重点是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作用。虽然 该建议仅限于这些纤维的体外分析，我们正在恒河猴中进行平行实验 猕猴来检查这些原纤维在体内的作用。我的计划在指导阶段， K99/R 00，如果得到资助，将开发淀粉样蛋白和生殖生物学的新技术技能， 应用这些技能来了解艾滋病毒的传播。这将通过以下方式实现： 与我的K99导师和咨询委员会密切合作。我的长期目标是增进我们对 围绕HIV在生殖器粘膜中传播的分子事件， 将这些知识用于开发新的艾滋病预防类别。

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.