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

描述（由申请方提供）：高灵敏度HIV RNA检测已证明，在血浆中病毒复制临床抑制的患者中，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，但目前还没有数据来评估这些部位药物分布的种间相似性或差异。这就造成了应用上的困难 从一个物种到另一个物种的研究发现，包括推断到人类。此外，尚未生成关于上述解剖组织内细胞内药物分布的信息。最后，有关这些位点药物分布和活性的因素信息有限，包括蛋白结合、药物转运活性和（在核苷类似物逆转录酶抑制剂的情况下）细胞内内源性核苷酸浓度。该项目的假设是：残留的活性病毒储库是抗逆转录病毒药物渗透到靶组织细胞亚型减少的结果，在设计合理的强化方案之前，必须彻底了解种属之间的组织药物分布。提出了三个具体目标：1）在涉及作为活性储库的组织中，表征来自5种药物类别的6种常用抗逆转录病毒药物在感染和未感染的人源化小鼠、非人灵长类动物和HIV+受试者的模型中的分布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，包括蛋白结合、细胞内内源性核苷酸浓度，和药物转运蛋白表达。3)开发一种新的IR-MALDESI-MSI方法，以可视化药物在所有三个物种的组织储库中的分散，并评估该技术在未来HIV感染和治疗组织研究中的作用。这项工作的目标是确定在细胞外和细胞内抗逆转录病毒药物的生物分布和消除活跃的HIV储库的功效中存在哪些种属差异和药理学障碍。从这一建议产生的数据将使合理选择的抗逆转录病毒药物非常适合于目标活性水库，确定药理学的优点和局限性的动物模型的HIV感染，并制定一个新的战略来衡量横截面药物在组织中的分散。

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.