Uncovering HIV/opioid effects in the brain at the single cell level: transcription, chromatin accessibility, and reservoir analysis in the SIV/cART/morphine/rhesus monkey model

在单细胞水平上揭示 HIV/阿片类药物对大脑的影响：SIV/cART/吗啡/恒河猴模型中的转录、染色质可及性和储库分析

• 批准号: 10220475
• 负责人: Shilpa J. Buch
• 金额: $ 141.74万
• 依托单位: UNIVERSITY OF NEBRASKA MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-08-15 至 2026-06-30
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10220475
• 关键词: Address; Affect; American; Animal Model; Animals; Anti-Retroviral Agents; Biological Assay; Biological Markers; Brain; Brain region; CD4 Positive T Lymphocytes; Cell Culture Techniques; Cells; Cellular Assay; Chromatin; Chronic; Controlled Study; Corpus striatum structure; Coupled; Critical Pathways; DNA; Data; Data Set; Disease; Drug abuse; Ensure; Epidemic; Experimental Designs; FAIR principles; Future; Gene Expression; Gene Expression Regulation; Generations; Genes; Genetic Transcription; Gold; HIV; HIV Infections; HIV-1; Human; Immune; Immunohistochemistry; In Situ Hybridization; Individual; Infection; Injections; Knowledge; Lymphocyte; Macaca mulatta; Mediating; Microglia; Modeling; Molecular; Morphine; National Institute of Drug Abuse; Neuroglia; Neurons; Neuropathogenesis; Opioid; Pathogenesis; Pathogenicity; Pathway interactions; Pharmaceutical Preparations; Phenotype; Population; Predisposition; Prefrontal Cortex; Primates; Protein Isoforms; RNA; Randomized; Reproducibility; Research; Role; SIV; Saline; Sample Size; Site; System; Systems Biology; Techniques; Therapeutic Intervention; Transposase; Twin Multiple Birth; Validation; Variant; Viral; Viral Genome; Viral Pathogenesis; Viral reservoir; Virus; Work; antiretroviral therapy; base; brain cell; cell type; clinically relevant; cocaine exposure; combinatorial; data mining; design; drug of abuse; epigenetic regulation; epigenomics; experimental group; experimental study; follow-up; insight; interdisciplinary approach; macrophage; member; methamphetamine exposure; monocyte; nonhuman primate; novel; opioid abuse; opioid epidemic; opioid exposure; opioid use; outreach; pandemic disease; power analysis; prevent; programs; response; simian human immunodeficiency virus; single cell technology; single-cell RNA sequencing; transcriptome; transcriptome sequencing; virology

Project Summary A comprehensive interdisciplinary approach proposed to support the Single Cell Opioid Responses in the Context of HIV (SCORCH) Program Expansion: CNS Data Generation for Chronic Opioid, Methamphetamine, and/or Cocaine Exposures RFA from the National Institute on Drug Abuse (NIDA). We will perform studies on the interplay of HIV pathogenesis and opioid abuse in the gold-standard SIV/nonhuman primate system, using single-cell RNA sequencing (scRNAseq) to assess individual cellular transcriptomes. This will give us obtain unparalleled insight into the viral and drug-induced effects on diverse cell types in brain in regions critical for HIV infection and opioid abuse. In addition, we will also focus on primary targets of HIV and drivers of neuropathogenesis, microglia and brain macrophages, using scRNAseq as well as single-cell Assay for Transposase-Accessible Chromatin sequencing (scATACseq) to gain insight into the epigenetic regulation of gene expression at the single cell level. These will be combined with state-of-the-art sensitive assays for virus in the brain. We will ensure our data adhere to the FAIR (Findable, Accessible, Interoperable, and Reusable) standards, and will work closely with other members of the SCORCH program to share and disseminate data to yield maximal advancement to the field. Our experiments are carefully designed to address the role of virus, drugs, and their interactions, as well as include the clinically relevant scenarios of those taking antiretrovirals to suppress virus, and those who do not. Our rigor and reproducibility are facilitated by our detailed power analysis with proper sample sizes, as well as our experimental design to match the standards of blocking, replication and randomization. The results from these studies will provide novel insights into the CNS effects of HIV and drug abuse.

项目总结