HIV-1 EnvLRS: a scalable, sequence-based assay for in-depth assessment of HIV-1 bNAb resistance

HIV-1 EnvLRS：一种可扩展、基于序列的检测方法，用于深入评估 HIV-1 bNAb 耐药性

• 批准号: 10324540
• 负责人: Gregory Michael Laird
• 金额: $ 30万
• 依托单位: ACCELEVIR DIAGNOSTICS, LLC
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-07-23 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10324540
• 关键词: AIDS clinical trial group; Acquired Immunodeficiency Syndrome; Address; Anti-Retroviral Agents; Antibodies; Archives; Base Sequence; Biological Assay; CD4 Positive T Lymphocytes; Cells; Chemistry; Clinical; Clinical Trials; Clonality; DNA; Data; Development; Diagnostic; Disease Progression; Dose; Dyes; Emulsions; Epitopes; Formulation; Foundations; Frequencies; Generations; Genes; Genetic Recombination; Genomic DNA; HIV; HIV Infections; HIV Seropositivity; HIV envelope protein; HIV-1; Immune system; Individual; Institutes; Left; Leukapheresis; Measurement; Microfluidics; Minor; Molecular; Participant; Performance; Peripheral Blood Mononuclear Cell; Persons; Pharmaceutical Preparations; Phase; Phenotype; Plasmids; Population; Recovery; Regimen; Reproducibility; Resistance; Retroviridae; Safety; Sampling; Sequence Analysis; Series; Small Business Innovation Research Grant; System; Testing; Time; Variant; Viral; Viral Vector; Virus Replication; Wisconsin; antiretroviral therapy; assay development; base; bioinformatics pipeline; clinical implementation; individual patient; neutralizing antibody; novel; novel strategies; novel therapeutic intervention; phase I trial; prediction algorithm; resistance mutation; side effect; single molecule; small molecule; success; virology

PROJECT SUMMARY/ABSTRACT Human immunodeficiency virus type-1 (HIV-1) is a retrovirus that infects CD4+ T cells of the immune system. If left untreated, people living with HIV-1 (PLWH) will progress to AIDS and may ultimately die as a result. Combination antiretroviral therapy (ART) with small-molecule drugs is extremely effective at stopping the replication of HIV-1 in infected individuals but requires daily dosing often with considerable side effects. Importantly, despite the success of this approach at suppressing HIV-1 replication to clinically undetectable levels, antiretroviral therapy is not curative. This is due to the persistence of HIV-1 in a silent, or latent, state within long-lived CD4+ T cells at extremely low frequencies. These latently infected cells are not targeted by current small-molecule ART regimens. As a result, PLWH must remain on lifelong antiretroviral therapy. Broadly neutralizing antibodies targeting critical epitopes in HIV-1 Env (HIV-1 bNAbs) are currently being developed as a potential approach to eliminate latent HIV-1 and/or as an alternative to small-molecule ART. HIV-1 bNAbs offer several advantages over traditional small-molecule ART, including the potential for long-acting formulations, reduced side effects, and the potential to eliminate latently infected cells over time. However, a major challenge to the implementation of HIV-1 bNAbs in treatment and cure is pre-existing variation or resistance in the bNAb-targeted epitopes. Scalable clinical tests are needed to determine (1) whether people who will receive HIV-1 bNAbs have pre-existing resistance, and (2) personalize HIV-1 bNAb combinations to each individual. To address this critical unmet need, AccelevirDx is developing the HIV-1 EnvLRS assay as the first scalable sequence-based test to assess HIV-1 bNAb resistance and predict antibody efficacy by in-depth env sequence analysis. Broadly, this proposal aims to (1) analytically qualify AccelevirDx’s novel, proprietary HiFi-dePCR approach for env amplification underlying HIV-1 EnvLRS, (2) determine the reproducibility of the HIV-1 EnvLRS assay of samples from PLWH, and (3) apply the assay to a recently completed ACTG A5340 clinical trial of the HIV-1 bNAb VRC01 in PLWH to assess assay performance and utility.

项目摘要/摘要 人类免疫缺陷病毒1型(HIV-1)是一种逆转录病毒，感染免疫的CD4+T细胞 系统。如果不治疗，艾滋病毒携带者(PLWH)将发展为艾滋病，并可能最终死于 结果。抗逆转录病毒疗法(ART)与小分子药物联合治疗对阻止 艾滋病毒-1在感染者身上复制，但需要每日剂量，往往有相当大的副作用。 重要的是，尽管这种方法成功地将HIV-1复制抑制到临床无法检测到 水平，抗逆转录病毒治疗是不能治愈的。这是由于HIV-1持续处于沉默或潜伏状态 在长寿命的CD4+T细胞中以极低的频率出现。这些潜伏感染的细胞不是 目前的小分子抗逆转录病毒疗法。因此，PLWH必须继续接受终身抗逆转录病毒治疗。 针对HIV-1env关键表位的广谱中和抗体(HIV-1 bNAbs)目前正在 被开发为消除潜伏的HIV-1和/或作为小分子的替代品的潜在方法 艺术。与传统的小分子抗逆转录病毒疗法相比，HIV-1 bNAbs具有几个优势，包括 长效配方，减少副作用，以及随着时间的推移消除潜伏感染细胞的可能性。 然而，在治疗和治疗中实施艾滋病毒-1bNAbs的一个主要挑战是预先存在的。 BNAb靶向表位的变异或抗药性。需要可扩展的临床测试来确定(1) 将接受HIV-1 bNAb的人是否具有预先存在的抵抗力，以及(2)个性化HIV-1 bNAb 每个个体的组合。为了解决这一关键的未得到满足的需求，AccelevirDx正在开发HIV-1 EnvLRS检测作为第一个可扩展的基于序列的测试来评估HIV-1bNAb耐药性并预测 通过深入的env序列分析获得抗体效价。总的来说，这项提议的目的是(1)分析合格 用于HIV-1 EnvLRS的新的、专有的高保真-去聚合酶链式反应方法，(2) 确定来自PLWH的样本HIV-1 EnvLRS检测的重复性，以及(3)将该检测应用于 最近完成的ACTG A5340临床试验对HIV-1bNAb VRC01在PLWH中的评估分析 性能和实用性。