Origin and Evolution of HIV-1 Drug Resistance in the RT-SHIVmne Macaque Model

描述（由申请人提供）：HIV-1耐药性严重影响感染者的治疗选择。建议使用至少含有两类药物的抗逆转录病毒联合疗法治疗HIV-1感染。通常使用2种核苷类似物逆转录酶（RT）抑制剂（NRTI）和1种非核苷RT抑制剂（NNRTI）的组合。在治疗过程中，HIV-1可以获得突变，从而逃避抗逆转录病毒药物的抑制。耐药病毒可以在宿主体内长期稳定，并可以水平或垂直传播给新的个体。然而，随着时间的推移，个体内耐药性的起源和持久性尚未深入研究。我们假设，耐药病毒将出现多灶性的组织，如粘膜部位，中枢神经系统和淋巴结，耐药之前，在血液中观察到，并将持续更长的时间相比，血液。我们还提出，抗逆转录病毒药物的药代动力学和药效学在这些解剖隔室的差异强烈影响耐药性的选择和持久性。 非人灵长类动物模型已被用于研究艾滋病的发病机制和免疫学，使用猿猴免疫缺陷病毒（SIV）。由于非核苷类逆转录酶抑制剂不能抑制SIV，因此不能在SIV模型中研究这些药物。因此，我们开发了一种猕猴模型来研究体内HIV-1耐药性。该模型使用感染由含有HIV-1HxB 2 RT编码区的SIVmne（RT-SHIVmne）组成的致病性嵌合病毒的猕猴。在具体目标1中，我们建议纵向检查耐药RT-SHIVmne在血液中和在NRTI或NNRTI单药治疗期间出现的不同解剖隔室中的出现。由于即使在抑制性抗逆转录病毒治疗期间，HIV-1也会持续存在于细胞和组织储库中，因此我们在具体目标2中提出，在停止单药治疗后和开始使用所有新药或包括单药治疗中使用的药物的抗逆转录病毒治疗方案后，确定野生型病毒重新出现的起源和耐药病毒的持续存在。为了探索耐药病毒出现的位点特异性差异是否可以通过不同的位点特异性药物浓度来解释，我们在具体目标3中建议检查治疗期间和治疗后血液和组织中抗逆转录病毒药物的药代动力学和药效学。 所提出的研究难以在人类中执行，但可以在我们的抗逆转录病毒治疗和耐药性的RT-SHIVmne模型中容易地进行。这些实验将为耐药性的解剖学起源、抗逆转录病毒治疗前后耐药性的持续性以及抗逆转录病毒药代动力学中位点特异性变异的影响提供新的见解。这项工作应有助于确定治疗艾滋病毒感染者的更好策略，特别是那些窝藏和潜在传播耐药病毒的人。 公共卫生相关性：本提案中的研究旨在确定宿主体内HIV-1耐药性的发展。我们希望预测耐药病毒将在哪里以及如何出现，这将有助于我们更有效地靶向治疗，以尽量减少耐药性的发展和持续。

DESCRIPTION (provided by applicant): HIV-1 drug resistance seriously impacts treatment options for infected individuals. It is recommended that HIV-1 infection be treated with combination antiretroviral therapy containing at least two classes of drugs. A combination of 2 nucleoside analog reverse transcriptase (RT) inhibitors (NRTIs) and 1 nonnucleoside RT inhibitor (NNRTI) is commonly used. During treatment, HIV-1 can acquire mutations that allow escape from inhibition by antiretroviral drugs. Drug resistant viruses can be stable within the host for long periods of time and can be transmitted horizontally or vertically to new individuals. However, the sites of origin and persistence of drug resistance within individuals over time has not been studied in depth. We hypothesize that drug resistant viruses will emerge multifocally in tissues, such as mucosal sites, the central nervous system, and lymph nodes, before resistance is seen in blood and will persist for longer periods of time as compared to blood. We also propose that differences in the pharmacokinetics and pharmacodynamics of antiretrovirals in these anatomic compartments strongly influence the selection and persistence of drug resistance. Nonhuman primate models have been useful in studying AIDS pathogenesis and immunology, using simian immunodeficiency virus (SIV). Because NNRTIs do not inhibit SIV, these drugs cannot be studied in the SIV model. Therefore, we developed a macaque model to study HIV-1 drug resistance in vivo. This model uses macaques infected with a pathogenic chimeric virus consisting of SIVmne containing the HIV-1HxB2 RT coding region (RT-SHIVmne). In Specific Aim 1, we propose to longitudinally examine the emergence of drug- resistant RT-SHIVmne in the blood and in different anatomical compartments that arise during NRTI or NNRTI monotherapy. Because HIV-1 persists in cellular and tissue reservoirs even during suppressive antiretroviral therapy, we propose in Specific Aim 2 to determine the origin of the re-emergence of wild-type virus and persistence of drug-resistant virus after discontinuation of monotherapy and after initiation of antiretroviral therapy using a regimen of all new drugs or one that includes the drug used in monotherapy in the infected animals. To explore whether site-specific differences in the emergence of drug resistant viruses are explained by different site-specific drug concentrations, we propose in Specific Aim 3 to examine the pharmacokinetics and pharmacodynamics of the antiretrovirals in the blood and tissues during and after therapy. The proposed studies are difficult to execute in humans but can be readily performed in our RT-SHIVmne model of antiretroviral therapy and drug resistance. These experiments will provide new insights into the anatomic origin of drug resistance, its persistence on and off antiretroviral therapy, and the influence of site- specific variation in antiretroviral pharmacokinetics. This work should help determine better strategies for treating HIV-infected individuals, particularly those harboring and potentially transmitting drug resistant virus. PUBLIC HEALTH RELEVANCE: The studies in this proposal are designed to determine where development of drug resistant HIV-1 occurs within the host. We hope to predict where and how resistant virus will emerge, which will help us to target therapy more effectively in an effort to minimize the development and persistence of drug resistance.