Mechanisms of Antiviral Drug Resistance, Host Factors and Retroviral Replication

抗病毒耐药机制、宿主因素和逆转录病毒复制

• 批准号: 8763523
• 负责人: VINAY K. PATHAK
• 金额: $ 25.4万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8763523
• 关键词: Affect; Amino Acids; Antiviral Agents; Antiviral Therapy; C-terminal; Cells; Clinical; DNA; DNA Integration; Drug resistance; Equilibrium; Excision; Goals; HIV Drug Resistance Program; HIV-1; Integration Host Factors; Mutation; Nucleosides; Nucleotides; Patients; Pharmaceutical Preparations; Phenotype; Production; Proteins; RNA Degradation; RNA-Directed DNA Polymerase; Reporting; Research; Resistance; Reverse Transcriptase Inhibitors; Reverse Transcription; Ribonuclease H; Site Visit; Viral; Viral Drug Resistance; Virus; non-nucleoside reverse transcriptase inhibitors; particle; uracil-DNA glycosylase; viral DNA

Most studies of clinical drug resistance to nucleoside and nonnucleoside reverse transcriptase (RT) inhibitors (NRTIs and NNRTIs, respectively) analyze the first 300 amino acids of RT and do not include the C-terminal connection subdomain (CN) or the RNase H domain (RH). We observed that several mutations in the CN and RH reduce RNase H activity, thereby affecting the balance between RNA degradation and nucleotide excision, and enhancing NRTI resistance. Our recent studies have demonstrated that mutations in the CN and the RH can also increase resistance to NNRTIs, and suggest a parallel mechanism by which resistance to both classes of RT inhibitors can be increased. It has been observed that CN mutations are present in drug-naive patients, and we are exploring the mechanisms by which CN mutations may be selected prior to initiation of antiviral therapy. While investigating the functional relevance of A3G localization to P bodies, we found that P body-associated protein Mov10 potently inhibits HIV-1 replication by reducing virus production and inhibiting reverse transcription. We are elucidating the mechanisms by which Mov10 inhibits HIV-1 particle production and reverse transcription. While examining the effects of A3G and A3F on viral DNA integration, we observed that uracil DNA glycosylase activity in the target cells, not the virus producer cells, results in nicking of the minus-strand DNA. We are exploring the significance of this unexpected phenotype to viral replication. [Corresponds to Pathak Project 3 in the October 2011 site visit report of the HIV Drug Resistance Program]

大多数对核苷类和非核苷类逆转录酶（RT）抑制剂（分别为NRTIs和NNRTIs）的临床耐药研究分析了RT的前300个氨基酸，不包括c端连接亚结构域（CN）或RNase H结构域（RH）。我们观察到CN和RH的一些突变降低了RNase H的活性，从而影响了RNA降解和核苷酸切除之间的平衡，增强了NRTI的抗性。我们最近的研究表明，CN和RH的突变也可以增加对NNRTIs的耐药性，并提出了一种平行的机制，通过这种机制可以增加对两类RT抑制剂的耐药性。据观察，CN突变存在于未接受药物治疗的患者中，我们正在探索CN突变在抗病毒治疗开始前可能被选择的机制。在研究A3G定位于P小体的功能相关性时，我们发现P小体相关蛋白Mov10通过减少病毒产生和抑制逆转录有效地抑制HIV-1复制。我们正在阐明Mov10抑制HIV-1颗粒产生和逆转录的机制。在检查A3G和A3F对病毒DNA整合的影响时，我们观察到尿嘧啶DNA糖基酶活性在靶细胞而不是病毒产生细胞中导致负链DNA的缺口。我们正在探索这种意想不到的表型对病毒复制的意义。【对应2011年10月HIV耐药项目实地考察报告中的Pathak Project 3】