Structural basis for activity of and resistance to HIV integrase inhibitors

HIV整合酶抑制剂的活性和耐药性的结构基础

基本信息

批准号：
10238819
负责人：
Dmitry Lyumkis
金额：
$ 66.85万
依托单位：
SALK INSTITUTE FOR BIOLOGICAL STUDIES
依托单位国家：
美国
项目类别：
财政年份：
2017
资助国家：
美国
起止时间：
2017-09-25 至 2022-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10238819
关键词：
Active Sites Address Adopted Affect Binding Biochemical Biological Biological Assay Biology Cells Chromatin Chromosomes Clinic Clinical Collaborations Communities Complement Complex Coupled Cryoelectron Microscopy DNA Data Development Disease Progression Drug Industry Drug Targeting Drug resistance Enzymes Future Generations Genes Guidelines HIV HIV Integrase HIV Integrase Inhibitors HIV-1 Histone H3 Hybrids Individual Infection Institutes Integrase Investigation Macromolecular Complexes Mediating Molecular Mutation Nucleoproteins Ohio Oligonucleotides Pathway interactions Patients Pattern Pharmaceutical Preparations Positioning Attribute Procedures Process Research Resistance Resistance development Resistance profile Resolution Role Scientist Spumavirus Structure Tail Techniques Translating United States National Institutes of Health Universities Variant Viral Virus Virus Integration Work antiretroviral therapy base clinically relevant design drug action drug development effective therapy experimental study improved inhibitor/antagonist insight interest lens epithelium-derived growth factor mutant novel particle profiles in patients prototype public health relevance response structural biology success tool transcriptional coactivator p75 viral DNA viral resistance virology

项目摘要

Abstract The Human Immunodeficiency Virus Type 1 (HIV-1, hereafter referred to as HIV) currently infects ~37 million people worldwide, and the number of infected individuals continues to rise. In the absence of a cure, antiretroviral therapy (ART) represents the primary treatment option, as it slows disease progression, and limits new infections. Integrase (IN) Strand Transfer Inhibitors (INSTIs) are a class of ART that block integration of viral DNA into host chromosomes, a process that is mediated by the viral IN enzyme, which assembles into oligomeric nucleoprotein complexes on the ends of viral DNA, termed “intasomes”. INSTIs selectively target intasomes and represent first-line therapies in the clinic. However, the emergence of IN variants resistant to INSTIs is a major clinical problem. Structural biology approaches can decipher the molecular mechanisms underlying drug action and resistance, providing useful information for rationally improving current therapies. In this proposal, approaches centered around revolutionary advances in cryo-electron microscopy for structural studies will be used to understand how INSTIs interact with their natural drug target, the HIV intasome, as well as mechanisms by which resistance to these drugs emerges. The proposed aims will address several major themes. Aim 1 will define the mechanisms of action of clinical and developmental INSTIs in the context of HIV intasomes. Procedures used to perform high-resolution structural studies of INSTI-bound complexes by cryo-EM will then be adopted to decipher novel, clinically relevant mechanisms of drug resistance that arise in response to INSTI treatment. This work will be complemented using biochemical and virology assays designed to dissect key interactions between HIV intasomes and INSTIs and to validate structural findings. Aim 2 will extend these findings to define INSTI mechanism of action in the presence of biologically relevant cellular factors, including methylated mononucleosomes (mMNs; the natural target for HIV integration) and lens epithelium-derived growth factor (LEDGF). Aim 2 will therefore define the biochemical and structural mechanisms by which INSTIs interact with and inhibit IN catalytic activity in the context of the intasome-LEDGF-mMN complex, thereby elucidating how INSTIs function in infected cells and their precise stage of activity. In addition to providing the first structural information for INSTI interactions with their natural drug target in the presence of relevant cellular factors, this work will: 1) elucidate how mutations within the IN active site disrupt drug binding, 2) define the precise stage and mechanism of action of this important class of drugs in a cellular context, and 3) provide blueprints for the rational improvement of future INSTIs. !

抽象的人类免疫缺陷病毒1型（HIV-1，以下称为HIV）当前感染了约3700万全球人民，受感染者的数量继续增加。在没有治疗的情况下抗逆转录病毒治疗（ART）代表了主要的治疗选择，因为它减慢了疾病的进展并限制了新的感染。整合酶（IN）链转移抑制剂（Instis）是一类艺术，将病毒DNA整合到宿主中染色体，该过程是由酶中病毒介导的，该过程聚集成寡聚核蛋白病毒DNA末端的复合物称为“ intomes”。有选择地针对静态并表示诊所中的一线疗法。然而，抗性研究的变体的出现是主要的临床问题。结构生物学方法可以破译药物作用的分子机制和阻力，为合理改善当前疗法提供有用的信息。在此提案中，方法以结构研究的冷冻电子显微镜的革命进步为中心了解研究所如何与其自然药物靶标相互作用，艾滋病毒Intasome以及通过的机制对这些药物的抗性出现。拟议的目标将解决几个主要主题。 AIM 1将定义临床作用机理并在艾滋病毒英特膜的背景下发展。用于执行高分辨率的程序然后，Cryo-Em对结合结合复合物的结构研究将被采用以破译小说在临床上响应于Insti治疗而产生的耐药性机制。这项工作将是使用旨在剖析HIV之间关键相互作用的生化和病毒学测定法完成 intomes and Instis并验证结构发现。 AIM 2将扩展这些发现以定义Insti 在存在生物学相关的细胞因子的情况下，作用机理，包括甲基化单核小体（MMN；艾滋病毒整合的自然靶标）和镜片上皮生长因子（LEDGF）。因此，AIM 2将定义Intist与Instis相互作用的生化和结构机制并抑制在Intasome-LeDGF-MMN复合物的背景下催化活性，从而阐明了如何 Instis在感染细胞及其活性阶段的功能。除了提供与其自然药物目标相互作用的第一个结构信息相关细胞因素的存在，这项工作将：1）阐明主动部位中的突变如何破坏药物结合，2）定义该重要类药物在细胞中的精确阶段和作用机理背景和3）为未来Intis的合理改进提供了蓝图。呢