Structural and Biochemical Effects of Capsid-targeting Molecules on HIV-1 Capsid Assembly

衣壳靶向分子对 HIV-1 衣壳组装的结构和生化影响

• 批准号: 10619783
• 负责人: WILLIAM MICHAEL MCFADDEN
• 金额: $ 4.77万
• 依托单位: EMORY UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-12-29 至 2026-12-28
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10619783
• 关键词: Acquired Immunodeficiency Syndrome; Adherence; Affect; Affinity; Anti-Retroviral Agents; Antiviral Agents; Antiviral resistance; Binding; Binding Proteins; Biochemical; Biological Assay; Biological Availability; Capsid; Capsid Proteins; Cells; Characteristics; Clinical; Complex; Cryoelectron Microscopy; Detection; Deuterium; Dipeptides; Dissociation; Drug Targeting; Drug resistance; Electron Microscopy; Epidemic; Future; Glycine; HIV; HIV Infections; HIV-1; Hydrogen; Hydrophobicity; In Vitro; Individual; Infection; Integrase; Integration Host Factors; Label; Life; Mass Spectrum Analysis; Medicine; Morphology; Multi-Drug Resistance; Mutation; Nature; Negative Staining; Nuclear Import; Optics; Patient Care; Patients; Pharmacotherapy; Phenotype; Phenylalanine; Proteins; RNA-Directed DNA Polymerase; Reporting; Research; Resistance; Resolution; Resources; Role; Scanning; Science; Site; Structure; Surface Plasmon Resonance; System; Therapeutic; Tube; Viral; Viral Drug Resistance; Viral Genome; Virion; Virus; Virus Replication; Visualization; analog; antiretroviral therapy; antiviral drug development; career; clinical practice; combat; design; drug resistant virus; experimental study; high risk; improved; insight; monomer; mutant; nanomolar; next generation; novel; novel strategies; particle; peptidomimetics; pressure; prevent; scaffold; skills; stoichiometry; success; therapy resistant; viral fitness

Abstract HIV infection impacts over 37 million individuals, with over 2/3 of these patients receiving antiretroviral therapies (ART). ART is sustained for a patient’s life and can lead to the emergence of drug-resistant HIV-1. To combat drug-resistance, an improved and diverse set of antiretrovirals are needed for clinical use. To this extent, the HIV-1 capsid is an excellent target for antiretroviral therapies as it has numerous, essential roles throughout the HIV-1 replication cycle. Compounds that target the capsid protein (CA), known as capsid effectors (CEs), offer a novel class of HIV-1 antivirals for potential clinical use. One CE with marked success is lenacapavir, developed by Gilead Sciences. Lenacapavir is exceptionally potent, however, early results show treatment with lenacapavir can cause the emergence of antiviral-resistant HIV-1. Our lab has previously reported highly potent antiretrovirals that target the same site as lenacapavir, within the FG-binding pocket. Compounds that bind to the FG-binding pocket of CA mimic a conserved phenylalanine-glycine (FG) dipeptide motif found in many host factors reported to bind CA. Here, I will characterize structural changes to the HIV-1 capsid upon treatment with highly potent CEs that bind the FG-binding pocket and calculate the biochemical parameters of CA•CE interactions for this class of antiretroviral therapeutics. The nature of CA•CE interactions will be assessed in wild-type (WT) and drug-resistant viruses to further our understanding of antiviral resistance. Electron microscopy (EM) will be used to visualize drug-resistant capsid assemblies and discern structural changes relative to WT assemblies (AIM 1). Rates of capsid assembly and changes to thermal stability upon drug-treatment will be calculated using assays designed to probe CA•CA interactions (AIM 2). These aims will study mutations that confer antiviral-resistance and results will be compared to WT CA to identify those with similar phenotypes and therefore higher risks of resistance. Further, CA•CE biochemical parameters of affinity and dissociation rates will be solved by label-free optical detection. This study will further our mechanistic understanding of compounds that bind to the FG-binding pocket of CA. Overall, these results will enable future research to strategically improve antiretrovirals, aiming to combat the HIV-1 epidemic.

抽象的 艾滋病毒感染影响超过3700万人，其中2/3例接受抗逆转录病毒疗法 （艺术）。艺术是为患者的生命而维持的，可以导致耐药HIV-1的出现。战斗 抗药性，临床使用需要改善和潜水的抗逆转录病毒。在这个程度上 HIV-1衣壳是抗逆转录病毒疗法的绝佳靶标，因为它在整个过程中具有许多重要的作用 HIV-1复制周期。靶向胶囊蛋白（CA）的化合物，称为衣壳效应（CES），提供 一种新型的HIV-1抗病毒药，用于潜在的临床用途。 Lenacapavir是一位获得明显成功的CE，开发了 吉利德科学。 Lenacapavir具有异常潜力，但是，早期结果显示了Lenacapavir的治疗 可能导致抗病毒HIV-1的出现。我们的实验室以前已经报道了高潜力的抗逆转录病毒 该位置与Lenacapavir相同的位置，在FG结合口袋内。结合FG结合的化合物 CA模拟苯丙氨酸 - 甘氨酸（FG）二肽基序中发现的许多宿主因子中发现 结合大约在这里，我将在高度有效的治疗后表征HIV-1 CAPSID的结构变化 结合FG结合口袋并计算CA的生化参数的CE•CE相互作用 抗逆转录病毒疗法类别。 CA的性质•CE相互作用将在野生型（WT）和 抗药性病毒，以进一步了解我们对抗病毒抗性的理解。电子显微镜（EM）将使用 可视化耐药的衣壳组件并辨别相对于WT组件的结构变化（AIM 1）。 在药物治疗时将使用CAPSID组装的速率和对热稳定性的变化，将使用测定法计算 旨在探测CA•CA相互作用（AIM 2）。这些目的将研究抗病毒抗病毒的突变 并将结果与​​WT CA进行比较，以识别具有相似表型的人，因此 反抗。此外，CA•亲和力和解离率的生化参数将通过无标签来解决 光学检测。这项研究将进一步我们对与FG结合结合的化合物的机械理解 大约的口袋。总体而言，这些结果将使未来的研究能够从战略上改善抗逆转录病毒，旨在 战斗HIV-1流行病。