Generating SEVI disaggregases to prevent HIV infection

描述(申请人提供)：人类免疫缺陷病毒1型(HIV-1)是获得性免疫缺陷综合征(AIDS)的病原体，已在全球感染约6000万人，并导致超过2500万人死亡。性传播是HIV-1感染的主要途径，最近在精液中发现了促进这一感染途径的因素。前列腺酸性磷酸酶片段是精液的主要成分，形成淀粉样纤维，与艾滋病毒病毒粒子结合，可以促进感染几个数量级，最高可达105倍。因此，一个潜在的预防策略是部署消除这些淀粉样蛋白形式的药物，这些淀粉样蛋白被称为精液衍生病毒感染增强因子(SEVI)纤维。不幸的是，淀粉样蛋白纤维是出了名的稳定，很难根除。在其他 在环境中，它们与各种致命的神经退行性疾病有关。然而，各种微生物已经将淀粉样蛋白的形成用于有益的目的，并且已经进化出可以快速逆转淀粉样蛋白形成的系统。一种天然蛋白质已经出现，它以前所未有的速度分解淀粉样纤维：蛋白质解聚酶Hsp104。HSP104能迅速溶解几种蛋白质的淀粉样蛋白，包括酵母蛋白Sup35和Ure2，以及在帕金森氏症中形成淀粉样纤维的？-突触核蛋白。我们假设，可以产生HSP104或SEVI优化的变体，以快速溶解或重塑SEVI原纤维，从而减少SEVI增强的艾滋病毒感染。因此，我们的目标是：(1)开发快速分解SEVI纤维的Hsp104变体；(2)确定分解后的产品是否降低了促进HIV感染的能力。这些研究将为开发SEVI解聚酶作为预防性制剂提供基础，最终目标是将它们合并到一种凝胶或溶液中，以溶解精液中的SEVI纤维并减少艾滋病毒的性传播。逆转艾滋病毒原纤维形成(而不是简单地抑制它)和阻断艾滋病毒的性传播的能力将提供对抗全球艾滋病毒/艾滋病大流行的强大和亟需的武器。我们以宿主蛋白构象(SEVI纤丝)为靶点的方法与以病毒为靶点的传统杀菌方法有根本不同，这一策略预计将与直接抗病毒策略协同作用。 与公共卫生相关：人类免疫缺陷病毒1型(HIV-1)是导致获得性免疫缺陷综合征(AIDS)的病毒，通过性传播，HIV-1感染通过人类精液中宿主蛋白形成的纤维增加约100,000倍。在这里，我们计划设计特定的酶来靶向并消除这些宿主蛋白纤维，从而阻止HIV-1感染。实现我们的目标将使迫切需要的预防艾滋病毒-1感染和防治全球艾滋病毒/艾滋病流行病的新战略成为可能。

DESCRIPTION (provided by applicant): Human immunodeficiency virus type-1 (HIV-1), the causative agent of acquired immune deficiency syndrome (AIDS), has infected ~60 million people worldwide and caused over 25 million deaths. Sexual transmission is the major route of HIV-1 infection and factors that promote this infectious route have recently been identified in semen. Fragments of prostatic acid phosphatase are a major component of semen and form amyloid fibrils that bind HIV virions and can promote infection by several orders of magnitude up to 105-fold. Therefore, a potential preventative strategy is to deploy agents that eliminate these amyloid forms, which are termed Semen-derived Enhancer of Virus Infection (SEVI) fibrils. Unfortunately, amyloid fibrils are notoriously stable and difficult to eradicate. In other settings, they are connected with various fatal neurodegenerative disorders. However, various microbes have harnessed the amyloid form for beneficial purposes, and systems have evolved that can rapidly reverse amyloid formation. One natural protein has emerged that resolves amyloid fibrils with unprecedented alacrity: the protein disaggregase, Hsp104. Hsp104 rapidly solubilizes amyloid forms of several proteins, including yeast prion proteins Sup35 and Ure2, as well as ?-synuclein, which forms amyloid fibrils in Parkinson's disease. We hypothesize that Hsp104 or SEVI-optimized variants can be generated to rapidly dissolve or remodel SEVI fibrils and thereby diminish SEVI-enhanced HIV infection. Thus, we aim to: (1) Develop Hsp104 variants that rapidly disassemble SEVI fibrils~ and (2) Determine whether disassembled products have diminished ability to promote HIV infection. These studies will provide the foundations for developing SEVI disaggregases as preventative agents with the ultimate goal of incorporating them into a gel or solution that dissolves SEVI fibrils in semen and reduces sexual transmission of HIV. The ability to reverse fibril formation (rather than simply inhibit it) and blck sexual transmission of HIV will provide a powerful and much needed weapon against the global HIV/AIDS pandemic. Our approach of targeting a host protein conformer (SEVI fibrils) is fundamentally different from traditional microbicidal approaches that target the virus, and this strategy is anticipated to synergize with direct antiviral strategies. PUBLIC HEALTH RELEVANCE: Human immunodeficiency virus type-1 (HIV-1), the virus that causes acquired immune deficiency syndrome (AIDS), is transmitted sexually and HIV-1 infection is enhanced approximately 100,000-fold by fibers formed by host proteins in human semen. Here, we plan to engineer specific enzymes that target and eliminate these host protein fibers and thereby block HIV-1 infection. Realization of our objectives will empower urgently needed new strategies to prevent HIV-1 infection and combat the global HIV/AIDS pandemic.