Mechanisms of HIV-1 Hypersensitivity to Islatravir

HIV-1 对 Islatravir 过敏的机制

• 批准号: 10731747
• 负责人: ALEXA Anne SNYDER
• 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: 2022-07-01 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10731747
• 关键词: 2' -deoxyadenosine; Action Potentials; Active Sites; Affect; Antiviral Agents; Binding; Biochemical; Biological Assay; Biology; Categories; Cells; Clinical Trials; Combined Modality Therapy; Complex; Computer software; DNA; Data; Development; Distant; Dose; Drug Combinations; Drug resistance; Effectiveness; Environment; Enzymes; Equipment; Excision; Future; HIV-1; Hydrophobicity; Hydroxyl Radical; Hypersensitivity; Individual; Kinetics; Lead; Methods; Molecular; Mutation; NNRTI-resistance; Nucleic Acids; Nucleosides; Patient Noncompliance; Patients; Pharmaceutical Preparations; Pharmacotherapy; Phase III Clinical Trials; Polymerase; Predisposition; Protocols documentation; RNA-Directed DNA Polymerase; Regimen; Resistance; Reverse Transcriptase Inhibitors; Role; Scientist; Site; Structure; Techniques; Testing; Training; Treatment Efficacy; Viral; Viral Drug Resistance; X-Ray Crystallography; antagonist; antiretroviral therapy; combat; compliance behavior; crosslink; design; drug resistance development; ds-DNA; experimental study; inhibitor; insight; mutant; non-nucleoside reverse transcriptase inhibitors; novel strategies; nucleoside analog; prevent; resistance mutation; synergism; treatment strategy; tripolyphosphate

PROJECT SUMMARY/ ABSTRACT As of 2020, 37.7 million patients are living with human immunodeficiency virus type 1 (HIV-1). Combination antiretroviral therapies (cARTs) are used to treat HIV-1 infected patients, but the effectiveness of these cARTs is threatened by viral drug resistance mutations most frequently caused by patient non-adherence to daily dosing regimens. Potent antivirals with long-acting potential are urgently needed to prevent drug resistance mutations. Islatravir (4’-ethynyl-2-fluoro-2’-deoxyadenosine), often referred to as EFdA, is a highly potent long-acting HIV- 1 nucleoside reverse transcriptase (RT) translocation inhibitor currently in phase III clinical trials. In these clinical trials, EFdA is administered in combination with other HIV-1 antivirals including non-nucleoside RT inhibitors. Interestingly, non-nucleoside RT inhibitor (NNRTI) treatment, which include Doravirine, Etravirine, Rilpivirine, or Elsulfavirine, leads to the RT F227C mutation, which remarkably makes HIV-1 more susceptible to EFdA. However, why a mutation located at the distant non-nucleoside RT inhibitor pocket would cause RT to become hypersensitive to EFdA, a compound that binds at the polymerase active site is not understood. Studies are needed to understand the mechanism of RTF227C hypersensitivity to EFdA and how this mutation affects HIV-1 susceptibility to various RT antivirals. I hypothesize that the F227C mutation at the NNRTI binding pocket structurally alters the polymerase active site, causing hypersensitivity to EFdA. To test this hypothesis, I will first determine the structural basis of RTF227C hypersensitivity to EFdA using X-ray crystallography (Aim 1). Second, I will determine the biochemical basis of RTF227C hypersensitivity to EFdA through a battery of biochemical assays. These assays will test how the F227C mutation impacts EFdA-triphosphate incorporation, the ability of RT to unblock EFdA-terminated primers, and the ability of RT to translocate along the nucleic acid (Aim 2). Finally, I will determine how the F227C mutation affects HIV-1 susceptibility to various EFdA and RT inhibitor combinations through biochemical and cell-based assays (Aim 3). I will take full advantage of our lab’s background in RT biology and role in the development of EFdA, as it provides the necessary environment, equipment, and streamlined protocols to complete these proposed experiments and contribute to my training. These planned studies will not only advance the understanding of how RT drug resistance mutations impart hypersensitivity to EFdA but will also aid in the development of optimized combinations with EFdA for the treatment of HIV-1.

项目总结/摘要 截至2020年，有3770万人感染人类免疫缺陷病毒1型（HIV-1）。组合 抗逆转录病毒疗法（cART）用于治疗HIV-1感染患者，但这些cART的有效性 受到病毒耐药性突变的威胁，最常见的原因是患者不依从每日给药 养生法迫切需要具有长效潜力的强效抗病毒药物来防止耐药性突变。 Islatravir（4 '-乙炔基-2-氟-2'-脱氧腺苷），通常称为EFdA，是一种高效长效HIV-1抑制剂。 1核苷逆转录酶（RT）易位抑制剂目前在III期临床试验。这些临床 在临床试验中，EFdA与其他HIV-1抗病毒药物（包括非核苷RT抑制剂）联合给药。 有趣的是，非核苷RT抑制剂（NNRTI）治疗，包括多拉韦林、依曲韦林、阿匹韦林或依曲韦林， Elsulfavirine导致RT F227 C突变，这显著地使HIV-1对EFdA更敏感。 然而，为什么位于远端非核苷RT抑制剂口袋的突变会导致RT成为 由于对EFdA超敏感，因此不了解在聚合酶活性位点结合的化合物。研究是 需要了解RTF 227 C对EFdA超敏反应的机制以及这种突变如何影响HIV-1 对各种RT抗病毒药物的敏感性。我假设NNRTI结合口袋的F227 C突变 在结构上改变聚合酶活性位点，引起对EFdA的超敏反应。为了验证这一假设，我将首先 使用X射线晶体学确定RTF 227 C对EFdA超敏的结构基础（目的1）。第二、 我将通过一系列生化试验来确定RTF 227 C对EFdA超敏反应的生化基础。 分析。这些测定将测试F227 C突变如何影响EFdA-三磷酸掺入， RT去阻断EFdA终止的引物，以及RT沿核酸沿着易位的能力（目的2）。 最后，我将确定F227 C突变如何影响HIV-1对各种EFdA和RT抑制剂的易感性 通过生物化学和基于细胞的测定（Aim 3），我会充分利用我们实验室 RT生物学背景和在EFdA开发中的作用，因为它提供了必要的环境， 设备和简化的协议来完成这些拟议的实验，并有助于我的培训。 这些计划中的研究不仅将促进对RT耐药突变如何影响 但也将有助于开发与EFdA的优化组合，用于治疗 治疗HIV-1。