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

基本信息

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
项目类别：
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

项目摘要

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.

项目摘要该应用程序的长期目标是研究CNS中持续存在并了解的HIV-1储层非法药物在建立和重新激活潜在储层中的作用。持续的潜在感染细胞在中枢神经系统位置是消除HIV-1的主要障碍。消除潜在水库的当前策略包括一种“冲击和杀伤”疗法，目的是外周血，仅占总回复者的1％。目前，是否可以设想消除HIV感染的CNS细胞的类似策略是未知。除了缺乏有关中枢神经系统HIV+细胞潜伏期的知识以及潜伏期的影响逆转药物，在感染HIV的人群中常见的非法药物构成了进一步的复杂性。许多已知非法药物会刺激HIV-1复制。由于当前测量外周血的方法 HIV储层不适用于固体组织或复制较差的细胞，例如巨噬细胞，小胶质细胞和大脑中发现的星形胶质细胞。我们已经开发了一种新颖的单细胞单个分子，多重分子，免疫荧光（IF）和RNA基于RNA的基于FISH的测定（SMIRA），以检测HIV-1的细胞。复制。使用自动化，可以扫描大量单元。在此提案中，我们将采用这本小说首先定量和表征细胞系模型中的延迟的方法，然后是大脑中的潜在储层小胶质细胞和星形胶质细胞，并描述三种滥用药物的作用（甲基苯丙胺，可卡因和大麻）关于潜在细胞形成的效率，潜在细胞的重新激活速率和建立跨脑障碍（BBB）的潜伏期。出于夏季本科课程的目的，我们将通过建立非法药物对潜在感染的中枢神经系统衍生细胞系的重新激活的作用潜伏期的体外模型。我们将采用永生的人类单核细胞和小胶质细胞系研究使用Smira的重新激活动力学和甲基甲基的作用。