Decay rate of archived HIV-1 drug resistance mutations

存档的 HIV-1 耐药突变的衰减率

• 批准号: 8603455
• 负责人: David Allenberg Katzenstein
• 金额: $ 7.85万
• 依托单位: STANFORD UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-05-17 至 2015-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8603455
• 关键词: AIDS clinical trial group; Address; Affect; Alleles; Antiviral Agents; Archives; Biological Assay; CD4 Lymphocyte Count; CD4 Positive T Lymphocytes; Chemistry; Clinical; Consensus; DNA; Data; Detection; Drug resistance; Enrollment; Exposure to; Failure; Frequencies; Future; Genotype; HIV; HIV Infections; HIV-1; Human Genome; Immune; Individual; Label; Lead; Life; Link; Measures; Methods; Minority; Mutation; NNRTI-resistance; Patients; Peptide Hydrolases; Peripheral Blood Mononuclear Cell; Pharmaceutical Preparations; Plasma; Point Mutation; Population; Prevention; Protease Inhibitor; Proviruses; RNA; RNA Sequences; RNA-Directed DNA Polymerase; Recording of previous events; Recycling; Regimen; Resistance; Rest; Sampling; Single Nucleotide Polymorphism; Technology; Testing; Time; Variant; Viral; Viral Load result; Viremia; Virus; antiretroviral therapy; base; deep sequencing; drug resistant virus; efavirenz; effective therapy; experience; innovation; next generation sequencing; non-nucleoside reverse transcriptase inhibitors; public health relevance; resistance mutation; sample fixation; transmission process; treatment strategy

DESCRIPTION: Antiretroviral therapy (ART) can reduce plasma HIV-1 levels below the limit of detection of clinical assays. However, incomplete suppression of virus can lead to rapid emergence of significant drug resistance mutations (DRM) that render certain drugs ineffective. The integration of drug resistant viruses into the host DNA and persistence in CD4+ cells in a latent state poses a major obstacle to effective treatment strategies and challenges new universal treatment paradigms for the eradication of HIV infection. The latent cellular reservoir functions as a historical archive of viral genotypes that have previously circulated in the host including provirus harboring DRM. However, little is known about the duration, persistence, and decay of DRM in the proviral reservoir over time. To address these questions, we identified subjects with known dates of virologic failure of Efavirenz-based regimens with emergence of drug resistant viremia (with the K103N mutation). These subjects were previously enrolled in ACTG trials but were subsequently treated with an effective second-line, boosted protease inhibitor-based regimen. CD4 count and viral load were measured and cryopreserved PBMC were collected every 6 months for up to 10 years. Following a failed first-line regimen, we hypothesize that consensus resistance mutations rapidly decay to levels below detection (<20%) in proviral DNA but persist in the latent reservoir with gradual decay over months to years. To test this hypothesis, our proposed study will 1) examine the frequency of DRM in proviral DNA following virologic failure and the detection of DRM in plasma RNA and 2) define the factors that affect fixation, retention, and decay over time and 3) use ultra-deep sequencing to quantify DRM over time in proviral DNA and establish the linkage between reverse transcriptase mutations. The information generated will have important implications for developing future treatment options for ART strategies to reduce clinical failure and mitigate the transmission of drug resistance. !

描述：抗逆转录病毒治疗 (ART) 可以将血浆 HIV-1 水平降低到临床检测的检测限以下。然而，病毒的不完全抑制可能导致显着的耐药突变（DRM）的迅速出现，从而使某些药物无效。耐药病毒整合到宿主 DNA 中并在 CD4+ 细胞中持续处于潜伏状态，这对有效治疗策略构成了重大障碍，并对根除 HIV 感染的新通用治疗范例提出了挑战。潜伏细胞库充当病毒基因型的历史档案，这些病毒基因型先前在宿主中传播，包括携带 DRM 的原病毒。然而，人们对原病毒库中 DRM 的持续时间、持久性和随时间的衰减知之甚少。为了解决这些问题，我们确定了已知日期的依非韦伦治疗方案病毒学失败并出现耐药病毒血症（带有 K103N 突变）的受试者。这些受试者之前曾参加过 ACTG 试验，但随后接受了有效的二线、基于蛋白酶抑制剂的加强方案治疗。每 6 个月测量一次 CD4 计数和病毒载量，并收集冷冻保存的 PBMC，持续长达 10 年。在一线治疗方案失败后，我们假设共识耐药突变会迅速衰减到前病毒 DNA 中检测到的水平 (<20%) 以下，但会持续存在于潜伏病毒库中，并在数月至数年内逐渐衰减。为了检验这一假设，我们提出的研究将1）检查病毒学失败后原病毒DNA中DRM的频率以及血浆RNA中DRM的检测，2）定义影响固定、保留和随时间衰减的因素，3）使用超深度测序来量化原病毒DNA中随时间推移的DRM并建立逆转录酶突变之间的联系。生成的信息将对制定 ART 策略的未来治疗方案具有重要意义，以减少临床失败并减轻耐药性的传播。 ！