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 HIV 感染影响了超过 3700 万人，其中超过 2/3 的患者正在接受抗逆转录病毒治疗 (ART) ART 会持续患者一生，并可能导致耐药 HIV-1 的出现。 由于耐药性，临床使用需要一套改进且多样化的抗逆转录病毒药物。 HIV-1 衣壳是抗逆转录病毒治疗的绝佳靶标，因为它在整个生命周期中发挥着众多重要作用。 HIV-1 复制周期的目标是衣壳蛋白 (CA)，称为衣壳效应子 (CE)。 lenacapavir 是一类具有潜在临床用途的新型 HIV-1 抗病毒药物。 吉利德科学公司生产的 Lenacapavi 非常有效，但早期结果显示 Lenacapavi 具有治疗作用。 可能导致出现抗病毒药物 HIV-1，我们的实验室此前曾报道过高效的抗逆转录病毒药物。 与 Lenacapavi 相同的位点，位于 FG 结合袋内，与 FG 结合的化合物结合。 CA 口袋模拟在许多报道的宿主因子中发现的保守苯丙氨酸-甘氨酸 (FG) 二肽基序 在这里，我将描述经过高效治疗后 HIV-1 衣壳的结构变化。 结合 FG 结合袋并计算 CA•CE 相互作用的生化参数的 CE CA•CE 相互作用的性质将在野生型 (WT) 和 将使用电子显微镜（EM）来进一步了解耐药病毒。 可视化耐药衣壳组件并辨别相对于 WT 组件的结构变化 (AIM 1)。 将使用测定法计算衣壳组装率和药物处理后热稳定性的变化 旨在探测 CA•CA 相互作用（AIM 2）。这些目标将研究赋予抗病毒耐药性的突变。 结果将与 WT CA 进行比较，以确定具有相似表型的结果，从而识别出具有更高风险的结果 此外，CA•CE的亲和力和解离率生化参数将通过无标记来解决。 这项研究将进一步加深我们对与 FG 结合的化合物的机理理解。 总体而言，这些结果将使未来的研究能够战略性地改进抗逆转录病毒药物，旨在 抗击 HIV-1 流行病。