Single Cell Analysis of HIV-1 Latent reservoir establishment in Humanized Mice

人源化小鼠体内 HIV-1 潜伏病毒库建立的单细胞分析

• 批准号: 10403201
• 负责人: Donald Doanman
• 金额: $ 4.66万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2021
• 资助国家: 美国
• 起止时间: 2021-11-24 至 2025-11-23
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10403201
• 关键词: Acute; Animal Model; CD4 Positive T Lymphocytes; Cause of Death; Cell Line; Cell Lineage; Cell surface; Cells; Complex; Cues; DNA; Data; Epidemic; Eukaryotic Initiation Factor-2; FRAP1 gene; Fluorescence; Gene Expression Profile; Gene Silencing; Generations; Genes; Genetic; Genetic Transcription; Goals; Grant; HIV; HIV Infections; HIV-1; Hour; In Vitro; Individual; Infection; Integration Host Factors; Kinetics; Knowledge; Laboratories; Light; Lymphoid Tissue; Maps; Memory; Modeling; Pathway interactions; Patients; Peripheral Blood Mononuclear Cell; Persons; Phenotype; Population Heterogeneity; Provirus Integration; Proviruses; Recording of previous events; Reporter; Reporting; Rest; Retroviridae; Role; Signal Transduction; Site; System; T memory cell; T-Lymphocyte; T-Lymphocyte Subsets; Technology; Testing; Time; Tissues; Viral; Viral Genes; Viral Load result; Viral Markers; Viral reservoir; Virus; Virus Integration; acute infection; antiretroviral therapy; base; cell type; chronic infection; effective therapy; experimental study; humanized mouse; in vivo; innovation; insight; integration site; latent HIV reservoir; mRNA Expression; medication compliance; new technology; novel; peripheral blood; single cell analysis; single-cell RNA sequencing; targeted treatment; tool; viral rebound

Abstract Human Immunodeficiency Virus (HIV) has infected 76 million people worldwide and caused the death of 38 million. Current treatments can reduce patient viral loads but fail to cure patients of infection. This is due to the ability of the virus to undergo latency during antiretroviral therapy. Latently infected cells are nearly indistinguishable from uninfected cells and there is no effective treatment to eliminate these cells. Our long- term goal is to describe at a single cell level, how early reservoir seeding occurs and the mechanisms that drive latency establishment. The proposed experiments in this grant are based on the hypothesis that during acute infection, a proportion of infected CD4+ T cells from diverse lineages of T cells such as resting CD4+ memory, activated T cells, naïve T cell, effector memory T cell undergo early latency and that transcriptional pathways such as mTOR and EIF2 signaling and provirus integration into transcriptionally repressive site are mechanisms of early latency establishment. This hypothesis is based on observations that: i) despite extremely early ART treatment, HIV latent reservoir may be established earlier than 72 hours after exposure ii) a diverse subset of T cells can harbor latent HIV iii) complex interactions between host and viral factors drive latency. First generation latency marking technology has been developed by the Chen lab to report the history of HIV infection through irreversible marking all HIV-infected cells called HIV-Induced Lineage Tracing (HILT). In this application, I propose a second-generation, innovative genetic marking technology, Enhanced HILT (EHILT), to further our knowledge the complexities of HIV latency. Using EHILT and validated small animal models of latency, I will define early latency kinetics in in vitro and in vivo acute and ART-treated infection using EHILT, identify transcriptional profiles and pathways associated with latency in vivo, and profile the integration sites of provirus with immediate latent phenotype.

抽象的 人类免疫缺陷病毒（HIV）已感染全球 7600 万人，并导致 38 人死亡 百万。目前的治疗可以减少患者的病毒载量，但无法治愈感染的患者。这是由于 病毒在抗逆转录病毒治疗期间经历潜伏期的能力。潜伏感染细胞几乎 与未感染的细胞无法区分，并且没有有效的治疗方法来消除这些细胞。我们的长期 术语目标是在单细胞水平上描述早期储库播种是如何发生的以及其机制 驱动延迟建立。本次资助中提出的实验基于以下假设： 急性感染，来自不同 T 细胞谱系的受感染 CD4+ T 细胞的比例，例如静息 CD4+ 记忆、活化 T 细胞、幼稚 T 细胞、效应记忆 T 细胞经历早期潜伏期，并且转录 mTOR 和 EIF2 信号传导以及原病毒整合到转录抑制位点等途径 早期潜伏期建立机制。该假设基于以下观察： i) 尽管极其 早期 ART 治疗中，HIV 潜伏库可能在暴露后 72 小时内建立 ii) 多样化 T 细胞子集可以隐藏潜伏的 HIV iii) 宿主和病毒因素之间复杂的相互作用驱动潜伏期。 陈实验室开发出第一代潜伏标记技术，用于报告艾滋病毒历史 通过不可逆地标记所有感染 HIV 的细胞（称为 HIV 诱导谱系追踪 (HILT)）来检测感染。在这个 应用中，我提出了第二代创新基因标记技术，增强型 HILT (EHILT)， 进一步了解艾滋病毒潜伏期的复杂性。使用 EHILT 和经过验证的小动物模型 潜伏期，我将使用 EHILT 定义体外和体内急性感染和 ART 治疗感染的早期潜伏期动力学， 识别与体内潜伏期相关的转录谱和途径，并分析整合位点 具有直接潜伏表型的原病毒。