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

探索精液淀粉样蛋白在促进 HIV 感染和受精中的作用

• 批准号: 8956470
• 负责人: Nadia R Roan
• 金额: $ 24.9万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-15 至 2016-11-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8956470
• 关键词: AIDS prevention; Accounting; Acquired Immunodeficiency Syndrome; Address; Advisory Committees; Amyloid; Amyloid Fibrils; Antiviral Agents; Artificial Insemination; Atomic Force Microscopy; Biochemical; Biological; Biological Assay; Cell physiology; Cells; Characteristics; Chemotaxis; Data; Development; Enhancers; Epidemic; Epithelial; Event; Female; Fertility Rates; Fertilization; Fertilization in Vitro; Funding; Future; Genetic Transcription; Genital system; Germ Cells; Goals; HIV; HIV Infections; HIV-1; Human; Immune; In Situ; In Vitro; Inflammation; Inflammatory; Integration Host Factors; Knowledge; Local Microbicides; Macaca mulatta; Measures; Mentors; Microscopy; Molecular; Molecular Conformation; Mouse Strains; Mucous Membrane; Mus; Neurobiology; Oocytes; Pattern recognition receptor; Persons; Pharmaceutical Preparations; Phase; Physiological; Process; Production; Property; Proteins; Recruitment Activity; Reproduction; Reproductive Biology; Research; Role; SIV; Seminal Plasma; Seminal fluid; Sexual Transmission; Signal Pathway; Signal Transduction; Techniques; Testing; Tissues; Translating; Viral; Virus; Virus Diseases; Work; abstracting; anti-HIV microbicide; antimicrobial; cellular targeting; chemokine; conformer; cytokine; design; egg; improved; in vivo; inhibitor/antagonist; insight; killings; male; microbicide; prevent; prophylactic; research study; response; sexual HIV transmission; skills; sperm cell; transmission process; vector

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.

7. 项目概要/摘要： 艾滋病毒最常在性接触后传播，精液是全球艾滋病毒的传播媒介 这种致命病毒的传播。我们的研究表明，精液远不是艾滋病毒的被动载体， 在体外显着增强 HIV 感染，我们进一步鉴定和表征了淀粉样原纤维 来自人类精液，可增加 HIV-1 与其细胞靶标的融合。这些原纤维可以增强艾滋病毒 感染超过几个数量级，因此可以作为开发 局部 HIV 杀菌剂。该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传染性及其对生殖器内细胞的影响 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 最有效增强艾滋病毒传染性的精液原纤维的形态特征。此信息 将揭示精液中淀粉样蛋白构象的类型，在设计特定的 精液中宿主因子的抑制剂。在目标 2 中，我将确定精液原纤维是否会诱发 宿主细胞的炎症，以及这在促进艾滋病毒感染中可能发挥的作用。据了解 炎症通常通过招募易感靶细胞并促进艾滋病毒传播来促进艾滋病毒传播 基因转录。了解精液原纤维在多大程度上导致宿主炎症 传播对于开发有效预防性传播的杀菌剂至关重要 病毒。最后，该提案的目标 3 侧重于更好地理解基本生理学 精液原纤维的功能。精液原纤维的进化并没有促进艾滋病毒感染，并且可能具有生物学意义 对人类的目的。有趣的是，HIV 与其细胞靶标的融合与 精子转化为卵子，提高了这些原纤维在受精过程中发挥作用的可能性。为了这个目标，我 提出体外受精（IVF）和体内人工授精实验以确定精液是否 我们表征的原纤维促进小鼠配子的融合。了解这些原纤维是否 促进受精的作用是开发艾滋病毒杀微生物剂的重要信息，并且还可以 在生殖领域产生重大影响。已投入大量精力 开发一种有效的艾滋病毒杀微生物剂。然而，该领域仍缺乏一种高效的药物 防止艾滋病毒的性传播，部分原因是我们对分子事件缺乏基本了解 周围粘膜HIV传播。该提案的重点是更好地了解艾滋病毒的一个方面 传播，即自然产生的精液原纤维增强艾滋病毒传染性的作用。虽然 该提案仅限于这些原纤维的体外分析，我们正在恒河猴中启动并行实验 猕猴来检查这些原纤维在体内的作用。我在指导阶段的计划 K99/R00，如果获得资助，将开发淀粉样蛋白和生殖生物学方面的新技术， 运用这些技能来了解艾滋病毒的传播。这将通过工作来完成 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的理解 围绕生殖器粘膜中艾滋病毒传播的分子事件，并将其转化为 将知识投入到新型艾滋病预防药物的开发中。