Multi-Species Mechanisms of Drug Bio-distribution in HIV Tissue Reservoirs

描述（由申请人提供）：高度灵敏的 HIV RNA 测定已证明，血浆中病毒复制受到临床抑制的患者体内病毒复制持续存在。许多解剖部位被认为是持续病毒产生的来源，包括胃肠道、中枢神经系统、淋巴组织和泌尿生殖系统。人源化小鼠和非人灵长类动物是 HIV 感染、持续存在和根除的模型，但目前尚无数据可评估这些部位药物分布的物种间相似性或差异。这导致申请困难 从一个物种到另一个物种的研究结果，包括推断到人类。此外，尚未生成有关上述解剖组织内细胞内药物分布的信息。最后，关于这些位点的药物分布和活性因素的信息有限，包括蛋白质结合、药物转运蛋白活性和（在核苷类似物逆转录酶抑制剂的情况下）细胞内内源核苷酸浓度。该项目的假设是：残留的活性病毒库是抗逆转录病毒对靶组织细胞亚型渗透减少的结果，在设计合理的强化方案之前，必须彻底了解物种之间的组织药物分布。提出了三个具体目标：1）在涉及活性储存库的组织中，描述感染和未感染的人源化小鼠、非人灵长类动物和 HIV 受试者模型中 5 类药物中 6 种常用抗逆转录病毒药物的分布特征。 2）研究可能负责抗逆转录病毒活性和分布的重要生理因素，包括蛋白质结合、细胞内内源核苷酸浓度和药物转运蛋白表达。 3) 开发一种新颖的 IR-MALDESI-MSI 方法，以可视化药物在所有三个物种的组织储存库中的分散情况，并评估该技术在未来 HIV 感染和治疗中组织研究中的作用。这项工作的目标是确定细胞外和细胞内抗逆转录病毒生物分布中存在哪些物种差异和药理学障碍以及消除活性艾滋病毒储存库的功效。该提案产生的数据将能够合理选择非常适合针对活性储存库的抗逆转录病毒药物，确定艾滋病毒感染动物模型的药理学优势和局限性，并开发一种测量组织中横截面药物分散的新策略。

DESCRIPTION (provided by applicant): Highly sensitive HIV RNA assays have demonstrated that viral replication persists in patients with clinically suppressed viral replication in their plasma. A number of anatomic sites have been proposed as the source of persistent viral production including the gastrointestinal tract, the central nervous system, lymphoid tissue, and the genitourinary system. Humanized mice and nonhuman primates are models for HIV infection, persistence, and eradication, yet no data currently exist to evaluate inter-species similarities or differences in drug distribution to these sites. This causes difficulty in applying research findings from one species to another, including extrapolating to humans. In addition, no information on intracellular drug distribution within the aforementioned anatomic tissues has been generated. Finally, there is limited information on factors responsible for drug distribution and activity at these sites, including protein binding, drug transporter activity, and (in the caseof nucleoside analogue reverse transcriptase inhibitors) intracellular endogenous nucleotide concentrations. The hypothesis for this project is: residual active viral reservoirs are a consequence of reduced antiretroviral penetration into target tissue cell subtypes, and a thorough understanding of tissue drug distribution between species must be conducted before rational intensification regimens can be designed. Three specific aims are proposed: 1) In tissues implicated as active reservoirs, characterize distribution of 6 commonly used antiretrovirals from 5 drug classes in models of infected and uninfected humanized mice, nonhuman primates, and HIV+ subjects 2) Investigate important physiologic factors that may be responsible for antiretroviral activity and distribution, including protein binding, intracellular endogenous nucleotide concentrations, and drug transporter expression. 3) Develop a novel IR-MALDESI-MSI approach to visualize the dispersion of drugs across tissue reservoirs for all three species, and assess the role of this technology for future tissue studies in HIV infection and therapy. The goal of this work is to identify what species differences and pharmacologic barriers exist in extracellular and intracellular antiretroviral biodistribution and efficacy in eliminating active HIV reservoirs. Data generated from this proposal will enable the rational selection of antiretrovirals well suited to target active reservoirs, determine the pharmacologic advantages and limitations of animal models of HIV infection, and develop a novel strategy to measure cross-sectional drug dispersion in tissues.