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.

摘要 艾滋病毒感染影响了3700多万人，其中三分之二以上的患者接受抗逆转录病毒治疗 （ART）。抗逆转录病毒疗法持续患者的一生，并可能导致耐药性HIV-1的出现。打击 为了克服耐药性，临床使用需要一套改进的和多样化的抗逆转录病毒药物。在这方面， HIV-1衣壳是抗逆转录病毒疗法的极好靶点，因为它在整个免疫过程中具有许多重要作用。 HIV-1复制周期。靶向衣壳蛋白（CA）的化合物，称为衣壳效应物（CE）， 一类具有潜在临床用途的新型HIV-1抗病毒药物。一个具有显著成功的CE是lenacapavir， 由吉利德科学公司提供。Lenacapavir是非常有效的，然而，早期结果显示， 会导致HIV-1产生抗药性。我们的实验室以前报道过高效的抗逆转录病毒药物 靶向与来那卡帕韦相同的位点，在FG结合口袋内。与FG结合蛋白结合的化合物 CA的口袋模拟了在许多宿主因子中发现的保守的苯丙氨酸-甘氨酸（FG）二肽基序 绑定CA。在这里，我将描述的结构变化的HIV-1衣壳治疗后，高效 结合FG结合口袋的CE，并为此计算CA·CE相互作用的生化参数。 抗逆转录病毒疗法的类别。CA·CE相互作用的性质将在野生型（WT）和 耐药病毒，以进一步了解抗病毒药物的耐药性。将使用电子显微镜（EM） 使耐药衣壳组装体可视化并辨别相对于WT组装体（AIM 1）的结构变化。 药物处理后衣壳组装速率和热稳定性变化将使用测定法计算 旨在探测CA·CA相互作用（AIM 2）。这些目标将研究赋予抗病毒药物抗性的突变 并将结果与WT CA进行比较，以确定具有相似表型的患者，因此具有更高的 阻力进一步地，CA·CE的亲和和解离速率的生化参数将通过无标记的 光学检测这项研究将进一步加深我们对与FG结合的化合物的机制的理解。 口袋CA总的来说，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， 抗击HIV-1流行病。