Dynamics of HIV Core Interactions

HIV核心相互作用的动态

• 批准号: 10650881
• 负责人: Stefan G Sarafianos
• 金额: $ 117.14万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-06-22 至 2027-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10650881
• 关键词: Antiviral Agents; Avidity; Behavior; Binding; Binding Proteins; Binding Sites; Biochemistry; Capsid; Capsid Proteins; Caring; Cell Nucleus; Cells; Complex; Computing Methodologies; Coupled; Cryoelectron Microscopy; Cytoplasm; Data; Dependence; Development; Drug Industry; Drug Targeting; Drug resistance; Ensure; Environment; Foundations; Future; Glycine; Goals; HIV; HIV-1; Hydrophobicity; Infection; Knowledge; Label; Lead; Mediating; Molecular; Molecular Biology; Nuclear; Nuclear Import; Nuclear Pore; Nuclear Pore Complex; Outcome; Patients; Peptides; Persons; Pharmaceutical Chemistry; Phase III Clinical Trials; Phenylalanine; Phosphotransferases; Prevention; Productivity; Proteins; Proteomics; Regulatory Pathway; Research; Resistance; Resistance profile; Route; Shapes; Site; Structure; Surface; Validation; Variant; Viral; Viral Drug Resistance; Virus; Virus-like particle; X-Ray Crystallography; activation-induced cytidine deaminase; base; biophysical techniques; cofactor; experience; frontier; improved; in silico; inhibitor; innovation; insight; live cell microscopy; molecular modeling; multidisciplinary; next generation; novel; novel therapeutics; protein protein interaction; resistance mechanism; screening; small molecule; small molecule inhibitor; structural biology; structural determinants; targeted treatment; therapeutic target; virology; virus core; virus host interaction

ABSTRACT, PROJECT 1 The overarching goals of the proposed studies are to i) elucidate dynamic, multifaceted interplay between HIV-1 and host cell that regulates infection and ii) define the virus-host interactions as therapeutic targets. The post-fusion journey of HIV-1 across the cytoplasm and via nuclear pore complex (NPC) is regulated by dynamic interactions of the conically shaped virus capsid shell with variety host proteins that either aid (dependency factors) or inhibit (restriction factors) infection. However, full identity and definitions of mechanisms of action of the cellular binding partners of HIV-1 capsid remain largely unknown. Furthermore, it is not clear how HIV-1 has evolved so that its capsid shell can bind selectively to diverse and seemingly unrelated regulatory host proteins that dictate the outcome of infection. In Aim 1, we will discover and characterize novel, dynamic interactions between HIV-1 capsid and host cell during virus ingress. Specifically, our efforts will focus on uncovering previously unrecognized, dynamic networks of cellular proteins that regulate HIV-1’s journey across the cytoplasm and through the NPC. Furthermore, we will elucidate structural determinants for selective and avid binding of these proteins to HIV-1 capsid. The hydrophobic capsid pocket, which engages several HIV-1 host dependency factors, has been successfully targeted by small molecule inhibitors. However, a relatively low barrier to resistance to current capsid targeting antivirals is a serious concern. In Aim 2, we will characterize interactions of existing and novel small molecule inhibitors with distinct sites on HIV-1 capsid. Specifically, we will define how known HIV-1 capsid inhibitors influence a dynamic structure of the conical HIV-1 capsid shell. Moreover, we will discover novel small molecules that bind at distinct sites on HIV-1 capsid and retain activity against viral variants resistant to current inhibitors. To accomplish above two aims we have assembled a highly collaborative B-HIVE team with complementary, multidisciplinary expertise in cryo-ET, cryo-EM, X-ray crystallography, HDX-MS, native MS, molecular modeling, live cell microscopy, virology, molecular biology, biochemistry, and medicinal chemistry. Our research will generate unprecedented insight into virus-host interactions that regulate HIV- 1’s journey during the virus ingress and define these interactions as the principal therapeutic target. Moreover, our structural and mechanistic characterization of multiple, distinct small molecule binding sites on the conical capsid surface will provide powerful means for ongoing efforts in pharmaceutical industry to rationally develop next generation of long-acting capsid inhibitors with enhanced barrier to resistance to transform care of the people living with HIV-1.

摘要，项目1 拟议研究的总体目标是：i）阐明以下方面之间动态的、多方面的相互作用： HIV-1和调节感染的宿主细胞，以及ii）将病毒-宿主相互作用定义为治疗靶点。 HIV-1通过细胞质和核孔复合物（NPC）的融合后旅程由以下调节： 锥形病毒衣壳壳与各种宿主蛋白质的动态相互作用， （依赖因素）或抑制（限制因素）感染。然而，完整的身份和定义 HIV-1衣壳的细胞结合配偶体的作用机制仍然是未知的。此外，委员会认为， 目前还不清楚HIV-1是如何进化的，以至于它的衣壳可以选择性地结合多种表面上看起来 决定感染结果的不相关调节宿主蛋白质。在目标1中，我们将发现和 表征病毒侵入期间HIV-1衣壳与宿主细胞之间的新型动态相互作用。 具体来说，我们的努力将集中在发现以前未被认识到的，动态的蜂窝网络， 调节HIV-1穿越细胞质和NPC的蛋白质。此外，我们将 阐明这些蛋白质与HIV-1衣壳选择性和亲和性结合的结构决定因素。的 与几种HIV-1宿主依赖性因子结合的疏水衣壳口袋已经成功地被 被小分子抑制剂靶向。然而，对当前衣壳的抗性的相对低的屏障 以抗病毒药物为目标是一个严重问题。在目标2中，我们将描述现有的和新的小分子之间的相互作用。 在HIV-1衣壳上具有不同位点的分子抑制剂。具体来说，我们将定义已知的HIV-1衣壳蛋白 抑制剂影响锥形HIV-1衣壳壳的动态结构。此外，我们将发现新的 在HIV-1衣壳上不同位点结合并保留抗病毒变异体活性的小分子 目前的抑制剂。为了实现上述两个目标，我们组建了一个高度协作的B-HIVE团队 在cryo-ET，cryo-EM，X射线晶体学，HDX-MS，native MS，分子建模，活细胞显微镜，病毒学，分子生物学，生物化学和医学 化学.我们的研究将对调节艾滋病毒的病毒-宿主相互作用产生前所未有的洞察力- 1的旅程在病毒入侵和定义这些相互作用作为主要的治疗目标。 此外，我们对多个不同的小分子结合位点的结构和机制表征， 将为制药工业的持续努力提供强有力的手段， 合理开发下一代长效衣壳抑制剂，增强对 改变对HIV-1感染者的护理。