Impact of Non - B HIV - 1 Subtype on second line Protease Inhibitor Regimens in Africa (INSPIRE)

非 B HIV-1 亚型对非洲二线蛋白酶抑制剂治疗方案的影响 (INSPIRE)

• 批准号: 10556406
• 负责人: Manhattan E Charurat
• 金额: $ 62.12万
• 依托单位: UNIVERSITY OF MARYLAND BALTIMORE
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-04-27 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10556406
• 关键词: Address; Adherence; Affect; Africa; Anti-Retroviral Agents; Antiretroviral drug resistance; Archives; Area; Binding; Biological Assay; Cameroon; Cells; Clinical; Cloning; Code; Coupled; Data; Drug resistance; Drug resistance pathway; Epidemiology; Evolution; Failure; Future; GAG Gene; Genes; Genetic; Genetic Polymorphism; Genetic Recombination; Genetic Variation; Genomic Segment; Genotype; Goals; HIV; HIV Infections; HIV drug resistance; HIV resistance; HIV-1; HIV-1 drug resistance; Human; In Vitro; Infection; Infrastructure; Integrase; Laboratories; Link; Longitudinal cohort; Measures; Mediating; Methods; Modification; Multi-Drug Resistance; Mutation; Nested Case-Control Study; Nigeria; Nigerian; Outcome; Pathway interactions; Patients; Pattern; Peptide Hydrolases; Pharmaceutical Preparations; Pharmacotherapy; Pharmacy facility; Phenotype; Plasma; Poland; Predictive Factor; Predisposition; Protease Inhibitor; Public Health; Publications; RNA; RNA-Directed DNA Polymerase; Recombinants; Regimen; Reporting; Research; Research Design; Research Personnel; Resistance; Risk; Role; Sampling; Site-Directed Mutagenesis; Specimen; System; Testing; Treatment Failure; Uganda; Variant; Viral; Viral Genome; Virus; Virus Replication; Yeasts; case control; clinically significant; cohort; design; drug resistant virus; env Genes; fitness; improved; inhibitor; mutant; next generation sequencing; non-nucleoside reverse transcriptase inhibitors; novel; pol genes; public health research; recombinant virus; resistance mutation; response; success; transmission process; virology

Abstract of research The finding that most patients with virologic failure (HIV-1 RNA >=1,000cp/mL) on second-line PI- containing regimens lack PI-resistance mutations in the protease (PR) gene is one of the main enigmas of antiretroviral drug resistance research. We hypothesized that env and gag sequences from these patients contain compensatory mutations, specifically in the Env, that confer PI resistance. A role of the gp41 CT in PI resistance may be linked to the role of virus maturation, triggered by PR, in activating Env fusion activity. In recent years, increasing numbers of clinical reports have observed failure of DTG-containing therapy in the absence of integrase (IN) mutations, suggesting that mutations outside IN may confer resistance in these patients. Our studies demonstrated the first instance of de novo selection of Env mutations that confer resistance to Dolutegravir (DTG) in vitro (unpublished data). We attribute this phenotype to the ability of the Env mutants to mediate highly efficient cell-to-cell transmission, resulting in an increase in the multiplicity of infection. In addition, up to 20-, 6- and 24-fold reduction in susceptibility to ATV, DRV and LPV, respectively, was observed in absence of PR mutations among HIV-1 CRF_02AG and subtype G infected patients. These findings have broad implications for our understanding of Env and Gag functions and the evolution of HIV-1 drug resistance. Our goal is to identify factors that predict virological failure (VF) on ATV/r or LPV/r as patients with VF on a PI-containing regimens have suboptimal future treatment and may be at an increased risk for developing integrase resistance inhibitor on a third-line regimen. A collaborative team involving investigators led by the Institute of Human Virology Nigeria will exploit robust longitudinal cohorts (Cameroon, Nigeria and Uganda) of patients on 2L regimens, coupled to well- characterized and archived clinical specimens that underpin a rigorous nested case-control study design. To test these interlocking hypotheses, we propose to: (i) use our newly developed HIV-xGen next generation sequencing method to extend understanding of viral evolution and HIV drug resistance across the viral genome and to determine whether there is effect modification by non-B HIV-1 subtypes, (ii) characterize recombinant viruses carrying mutations identified in Aim 1, using established phenotypic assays to determine their effect on drug susceptibility and site-directed mutagenesis, and lastly (iii) determine differences in replicative fitness on drug susceptibility with use of a yeast recombination-based cloning system. Our plans to study these patients and their viruses (subtype A, C, D, G, and CRF02_AG, CRF43_02G, HIV-1 O and HIV-1 N) provides a unique opportunity to determine the phenotypic and clinical significance of genotypic changes in HIV-1 gag, gag-pol and env genes. All in all, this proposal will permit exciting epidemiological, clinical, and public health research opportunities.

研究摘要 研究发现，大多数二线 PI 治疗失败的患者（HIV-1 RNA >=1,000cp/mL） 含有蛋白酶 (PR) 基因中缺乏 PI 抗性突变的方案是主要的原因之一 抗逆转录病毒药物耐药性研究的谜团。我们假设 env 和 gag 序列 来自这些患者的补偿性突变，特别是在 Env 中，赋予 PI 反抗。 gp41 CT 在 PI 抗性中的作用可能与病毒成熟的作用有关， 由 PR 触发，激活 Env 融合活性。近年来，临床上越来越多 报告观察到在缺乏整合酶 (IN) 的情况下含 DTG 的治疗失败 突变，表明 IN 之外的突变可能会给这些患者带来耐药性。我们的 研究证明了 Env 突变的从头选择的第一个实例，该突变赋予 体外对多替拉韦 (DTG) 的耐药性（未发表的数据）。我们将这种表型归因于 Env 突变体介导高效细胞间传播的能力，从而导致 感染复数增加。此外，还可以减少高达 20、6 和 24 倍 在没有 PR 突变的情况下，观察到分别对 ATV、DRV 和 LPV 的易感性 HIV-1 CRF_02AG 和 G 亚型感染患者中。这些发现具有广泛的 对我们理解 Env 和 Gag 功能以及 HIV-1 药物进化的影响 反抗。我们的目标是确定预测 ATV/r 或 LPV/r 病毒学失败 (VF) 的因素： 接受含 PI 治疗方案的 VF 患者未来治疗效果不佳，可能处于 在三线治疗方案中出现整合酶耐药抑制剂的风险增加。一个协作的 由尼日利亚人类病毒学研究所领导的研究人员组成的团队将利用强大的 采用 2L 方案的患者的纵向队列（喀麦隆、尼日利亚和乌干达），加上良好的 表征和存档的临床标本支持严格的巢式病例对照研究 设计。 为了检验这些相互关联的假设，我们建议：(i) 接下来使用我们新开发的 HIV-xGen 世代测序方法扩展对病毒进化和艾滋病毒耐药性的理解 跨病毒基因组并确定是否存在非 B HIV-1 的效应修饰 亚型，(ii) 表征携带目标 1 中确定的突变的重组病毒，使用 建立表型测定以确定其对药物敏感性和位点定向的影响 诱变，最后（iii）确定复制适应性对药物敏感性的差异 使用基于酵母重组的克隆系统。我们计划研究这些患者及其 病毒（A、C、D、G 亚型和 CRF02_AG、CRF43_02G、HIV-1 O 和 HIV-1 N）提供 确定基因型变化的表型和临床意义的独特机会 HIV-1 gag、gag-pol 和 env 基因。总而言之，这项提案将带来令人兴奋的流行病学、 临床和公共卫生研究机会。