Effect of Drugs of Abuse on CNS HIV-1 Reservoirs and Neuropathogenesis

滥用药物对中枢神经系统 HIV-1 病毒库和神经发病机制的影响

• 批准号: 10419775
• 负责人: GANJAM V KALPANA
• 金额: $ 0.95万
• 依托单位: ALBERT EINSTEIN COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2022-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10419775
• 关键词: AIDS/HIV problem; Anatomy; Anti-Retroviral Agents; Astrocytes; Automation; Autopsy; Biological Assay; Biological Models; Blood - brain barrier anatomy; Brain; CD4 Positive T Lymphocytes; Cannabis; Cell Culture Techniques; Cell Line; Cells; Cocaine; Consumption; Development; Drug Screening; Future; Genetic Transcription; Goals; HIV; HIV-1; Human; Illicit Drugs; Immunofluorescence Immunologic; In Vitro; Infection; Kinetics; Knowledge; Measures; Methamphetamine; Methods; Microglia; Modeling; Nature; Neuraxis; Neuropathogenesis; Patients; Population; Proteins; Proviruses; RNA; Rest; Role; Sampling; Scanning; Shock; Solid; Time; Tissues; Viral; Viral Load result; Virus; base; cell type; drug of abuse; established cell line; in vitro Model; macrophage; memory CD4 T lymphocyte; methamphetamine effect; methamphetamine use; monocyte; novel; patient population; peripheral blood; prevent; programs; reactivation from latency; response; single molecule; specific biomarkers; tool; undergraduate student

PROJECT SUMMARY The long term goal of this application is to study HIV-1 reservoirs that persist in CNS and to understand the role of illicit drugs in establishing and reactivating the latent reservoirs. Persistence of latently infected cells in CNS poses a major barrier for HIV-1 eradication. Current strategies to eliminate the latent reservoirs include a “shock and kill” therapy and is aimed at peripheral blood, which constitutes only 1% of the total reservoirs. Whether it is possible to envision similar strategies for the eradication of HIV-infected CNS cells is currently unknown. In addition to the lack of knowledge about latency in CNS HIV+ cells and the effects of latency reversing agents, illicit drugs common within the HIV-infected populations constitute a further complexity. Many illicit drugs are known to stimulate HIV-1 replication. Since the current method to measure the peripheral blood HIV reservoir is not applicable to solid tissues or to cells that replicate poorly such as macrophages, microglia and astrocytes found in the brain. We have developed a novel, Single cell-single molecule, Multiplex, Immunofluorescence (IF) and RNA FISH-based Assay (SMIRA) to detect cells in which HIV-1 is actively replicating. Using automation, a large number of cells can be scanned. In this proposal, we will employ this novel method to first quantitate and characterize latency in cell line models, then the latent reservoirs in brain derived microglia and astrocytes and delineate the effect of three drugs of abuse (methamphetamine, cocaine and cannabis) on the efficiency of formation of latent cells, the rate of reactivation of latent cells, and establishment of latency across blood-brain-barrier (BBB). For the purpose of the summer undergraduate program, we will be studying the role of illicit drugs on the reactivation of latently infected CNS-derived cell lines by establishing in vitro models of latency. We will employ immortalized human monocyte and microglial cell lines to study the kinetics of reactivation and the effect of METH using SMIRA.

项目总结 这项应用的长期目标是研究持续存在于中枢神经系统的HIV-1宿主，并了解 非法药物在建立和重新激活潜在储藏库方面的作用。潜伏感染细胞的持久性 在CNS的传播对根除HIV-1构成了主要障碍。目前消除潜在油气藏的策略包括 这是一种“休克和杀死”疗法，针对的是外周血液，而外周血液只占总储存库的1%。 目前，是否有可能设想类似的策略来根除感染艾滋病毒的中枢神经系统细胞 未知。除了对中枢神经系统HIV+细胞潜伏期和潜伏期的影响缺乏了解外 逆转因素、艾滋病毒感染人群中常见的非法药物构成了进一步的复杂性。许多 众所周知，非法药物会刺激艾滋病毒-1复制。由于目前测量外周血的方法 HIV库不适用于固体组织或复制能力较差的细胞，如巨噬细胞、小胶质细胞 以及大脑中发现的星形胶质细胞。我们已经开发出一种新颖的，单细胞-单分子，多重， 免疫荧光(IF)和基于RNA FISH的分析(Smira)检测HIV-1活跃的细胞 复制。使用自动化技术，可以扫描大量细胞。在这个提案中，我们将使用这部小说 方法首先量化和表征细胞系模型中的潜伏期，然后推导出大脑中的潜伏库 小胶质细胞和星形胶质细胞，并描绘了三种滥用药物(甲基苯丙胺、可卡因和 大麻)对潜伏细胞的形成效率、潜伏细胞的复活率和建立 血脑屏障(BBB)的潜伏期。为了暑期本科课程的目的，我们将 研究非法药物对潜伏感染的中枢神经系统来源的细胞系重新激活的作用 潜伏期的体外模型。我们将使用永生化的人类单核细胞和小胶质细胞系来研究 使用Smira的再激活动力学和冰毒的影响。