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 拟议的研究的总体目标是i）阐明动态，多方面的相互作用 HIV-1和调节感染的宿主细胞，ii）将病毒宿主相互作用定义为治疗靶标。 HIV-1跨细胞质和核孔复合物（NPC）的融合后旅程受到 带有多种宿主蛋白​​的预期形状病毒capsid壳的动态相互作用，这要么有助于 （依赖性因素）或抑制（限制因素）感染。但是，完全身份和定义 HIV-1衣壳的细胞结合伴侣的作用机制仍然很大未知。此外， 目前尚不清楚HIV-1是如何发展的，因此它的衣壳外壳可以选择性地结合到多样性和看似 决定感染结果的无关调节宿主蛋白。在AIM 1中，我们会发现并 表征病毒入学期间HIV-1衣壳和宿主细胞之间的新型动态相互作用。 特别是，我们的努力将集中于揭示先前未知的细胞动态网络 调节HIV-1穿越细胞质和NPC的蛋白质。此外，我们会的 阐明这些蛋白质与HIV-1衣壳的选择性和狂热结合的结构确定剂。这 与多个HIV-1宿主依赖性因素相关的疏水性衣壳袋已成功成功 由小分子抑制剂靶向。但是，对电流capsid的阻力相对较低 靶向抗病毒药是一个严重的问题。在AIM 2中，我们将表征现有和新颖的小型相互作用 HIV-1衣壳上具有不同位点的分子抑制剂。具体而言，我们将定义如何知道HIV-1 CAPSID 抑制剂会影响锥形HIV-1衣壳壳的动态结构。而且，我们会发现小说 小分子在HIV-1衣壳上的不同位点结合并保持对病毒变异的活性抗性 到当前的抑制剂。为了实现以上目标，我们组建了一个高度协作的B-HY团队 具有Cryo-ET，Cryo-EM，X射线晶体学，HDX-MS的完整，多学科专业知识 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。我们的研究将产生对调节HIV-的病毒宿主相互作用的前所未有的见解。 1在病毒中的旅程，并将这些相互作用定义为主要治疗靶点。 此外，我们对多个不同的小分子结合位点的结构和机械表征 在圆锥形的衣壳表面将为制药行业的持续努力提供强大的手段 理性地发展下一代长效封闭抑制剂，具有增强的抵抗力的障碍 转变对HIV-1的人们的护理。