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

摘要 艾滋病毒感染影响了3700万人，其中超过三分之二的患者接受了抗逆转录病毒治疗 (艺术)。抗逆转录病毒治疗将持续患者的一生，并可能导致抗药性HIV-1的出现。去战斗 耐药，临床需要一套改进的和多样化的抗逆转录病毒药物。从这个意义上说， HIV-1衣壳蛋白是抗逆转录病毒治疗的极佳靶点，因为它在整个 HIV-1复制周期。以衣壳蛋白(CA)为靶标的化合物，称为衣壳效应器(CES)，提供 一类潜在临床应用的新型HIV-1抗病毒药物。一种取得显著成功的CE是莱那帕韦，已开发 作者：Gilead Sciences。然而，早期的研究结果显示，来那帕韦的治疗效果非常显著。 可导致出现具有抗病毒耐药性的HIV-1。我们的实验室此前曾报道过高效抗逆转录病毒药物 靶向与来那帕韦相同的部位，在FG结合口袋内。与FG结合的化合物 CA的Pocket模拟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 来绑定CA。在这里，我将描述在高度有效的治疗后艾滋病毒-1衣壳的结构变化 CES结合FG结合口袋并计算CA·CE相互作用的生化参数 一类抗逆转录病毒治疗药物。CA·CE相互作用的性质将在野生型(WT)和 抗药性病毒进一步加深了我们对抗病毒耐药性的理解。将使用电子显微镜(EM) 可视化耐药衣壳组件并辨别与WT组件相关的结构变化(目标1)。 药物治疗时衣壳的组装速率和热稳定性的变化将使用化验来计算 旨在探索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。 并将结果与WT CA进行比较，以确定那些表型相似的人，因此风险更高 抵抗。此外，CA·CE的亲和力和解离率等生化参数将通过无标记的方法求解 光学探测。这项研究将进一步加深我们对FG结合化合物的机理的理解 加州的口袋。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疫情。

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.