Mechanisms of HIV-1 Hypersensitivity to Islatravir

HIV-1 对 Islatravir 过敏的机制

基本信息

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
项目类别：
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&apos -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

项目摘要

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.

项目概要/摘要截至 2020 年，有 3770 万名人类免疫缺陷病毒 1 型 (HIV-1) 感染者。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坚持每日给药迫切需要具有长效潜力的强效抗病毒药物来预防耐药突变。 Islatravir（4'-乙炔基-2-氟-2'-脱氧腺苷），通常称为 EFdA，是一种高效长效的 HIV- 1 核苷逆转录酶（RT）转位抑制剂目前处于 III 期临床试验中。试验中，EFdA 与其他 HIV-1 抗病毒药物（包括非核苷 RT 抑制剂）联合使用。建议使用非核苷 RT 抑制剂 (NNRTI) 治疗，包括多拉韦林 (Doravirine)、依曲韦林 (Etravirine)、利匹韦林 (Rilpivirine) 或 Elsulfavirine 会导致 RT F227C 突变，从而使 HIV-1 更容易受到 EFdA 的影响。然而，为什么位于远处非核苷 RT 抑制剂口袋的突变会导致 RT 变成对 EFdA 过敏，一种与聚合酶活性位点结合的化合物尚不清楚。需要了解 RTF227C 对 EFdA 过敏的机制以及这种突变如何影响 HIV-1 我认为 NNRTI 结合口袋处存在 F227C 突变。结构上改变了聚合酶活性位点，导致对 EFdA 过敏。为了检验这一假设，我将首先进行测试。使用 X 射线晶体学确定 RTF227C 对 EFdA 过敏的结构基础（目标 1）。我将通过生化电池确定 RTF227C 对 EFdA 过敏的生化基础这些测定将测试 F227C 突变如何影响 EFdA-三磷酸掺入的能力。 RT 解封 EFdA 终止的引物，以及 RT 沿着核酸易位的能力（目标 2）。最后，我将确定 F227C 突变如何影响 HIV-1 对各种 EFdA 和 RT 抑制剂的敏感性通过生化和细胞分析进行组合（目标 3）。 RT 生物学背景以及在 EFdA 发展中的作用，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为我的培训做出贡献。这些计划中的研究不仅将增进对 RT 耐药突变如何传递的理解对 EFdA 过敏，但也有助于开发与 EFdA 的优化组合，用于 HIV-1 的治疗。