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 大多数病毒学失败（HIV-1 RNA >=1，000 cp/mL）的患者在二线PI-1治疗后， 含有方案缺乏蛋白酶（PR）基因中的PI抗性突变是主要的 抗逆转录病毒药物耐药性研究之谜。我们假设env和gag序列 来自这些患者的基因含有补偿性突变，特别是在Env中， 阻力gp 41 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，越来越多的临床 有报道观察到在不存在整合酶（IN）的情况下含DTG的治疗失败 突变，表明IN以外的突变可能会在这些患者中产生耐药性。我们 研究表明，第一个从头选择Env突变的例子， 体外对度鲁特韦（DTG）的耐药性（未发表数据）。我们将这种表型归因于 Env突变体介导高效细胞间传递的能力，导致 感染的多重性增加。此外，可将细胞中的 在没有PR突变的情况下，分别观察到对ATV、DRV和LPV的敏感性 HIV-1CRF_02AG和G亚型感染者中。这些发现具有广泛的 对我们理解Env和Gag功能以及HIV-1药物进化的意义 阻力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方案治疗的VF患者未来治疗效果欠佳， 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 由尼日利亚人类病毒学研究所领导的研究小组将利用强大的 接受2L方案的患者纵向队列（喀麦隆、尼日利亚和乌干达）， 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 为了验证这些相互关联的假设，我们建议：（i）使用我们新开发的HIV-xGen next 一代测序方法扩展了对病毒进化和HIV耐药性的理解 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中鉴定的突变的重组病毒，使用 建立了表型分析，以确定其对药物敏感性的影响， 诱变，最后（iii）确定复制适合度对药物敏感性的差异， 使用基于酵母重组的克隆系统。我们研究这些病人和他们的 病毒（亚型A、C、D、G和CRF02_AG、CRF43_02G、HIV-1 O和HIV-1 N）提供了一种 独特的机会，以确定表型和临床意义的基因型变化， HIV-1 gag、gag-pol和env基因。总而言之，这项提案将允许令人兴奋的流行病学， 临床和公共卫生研究机会。