Targeting HIV-1 RNA modifications in latently infected CD4+ T cells for therapeutic development

针对潜伏感染 CD4 T 细胞中的 HIV-1 RNA 修饰进行治疗开发

基本信息

批准号：
10596144
负责人：
Li Wu
金额：
$ 69.99万
依托单位：
UNIVERSITY OF IOWA
依托单位国家：
美国
项目类别：
财政年份：
2022
资助国家：
美国
起止时间：
2022-04-01 至 2025-03-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10596144
关键词：
Address Adult Antiviral Agents CD4 Positive T Lymphocytes Cell Line Cells Critical Pathways Development Drug Targeting Funding Opportunities Gene Expression Genetic Transcription HIV HIV 1 Envelope Protein gp120 HIV-1 HIV/AIDS Immune response In Vitro Individual Infection Innate Immune Response Knock-out Knowledge Macrophage Memory Modification Natural Immunity Pathway interactions Patients Peripheral Blood Mononuclear Cell Phase Play Post-Transcriptional RNA Processing Post-Transcriptional Regulation Proteins Proviruses RNA Reader Regulation Reporting Rest Role T-Lymphocyte Subsets Testing Therapeutic Translating Up-Regulation Viral Viral reservoir Virus Diseases Virus Latency Virus Replication antiretroviral therapy cellular targeting design epitranscriptomics in vivo inhibitor knock-down latent HIV reservoir multidisciplinary novel novel strategies novel therapeutic intervention posttranscriptional power analysis small molecule therapeutic development therapeutic target transcriptome sequencing

项目摘要

Project Summary/Abstract Despite highly effective anti-retroviral therapy (ART), the latent HIV-1 reservoir in resting CD4+ T cells is the major barrier to a functional cure of HIV-1 infection. Our overall objectives of this R61/R33 bi-phasic project are: to understand the mechanisms of post-transcriptional regulation of HIV-1 RNA in HIV-1-infected individuals who are on ART (Exploratory R61 phase), and to develop a novel therapeutic strategy to alter RNA post- transcriptional modifications as a potential therapeutic platform for inhibiting HIV-1 replication (Developmental R33 phase). Our multidisciplinary group is uniquely poised to address several key questions highlighted in this funding opportunity. My lab was among three groups that independently discovered that N6-methyladenosine (m6A) modifications of HIV-1 RNA modulate viral replication in CD4+ T cells in vitro. Using CD4+ T cell lines and primary CD4+ T cells from healthy donors, we investigated the mechanisms by which m6A modifications modulate HIV-1 infection. We also found that m6A modifications of HIV-1 RNA inhibit innate antiviral immune responses in primary macrophages from healthy donors. Our in vitro studies suggested that m6A modifications of HIV-1 RNA play a critical role in viral replication and innate immune responses to viral infection. However, the role of m6A modifications of HIV-1 RNA in regulating viral replication in HIV-1-infected individuals on ART remains unknown. We aim to fill this important knowledge gap and to translate the findings into potential anti- HIV-1 therapeutics. We hypothesize that m6A modifications of HIV-1 RNA help establish and maintain viral latency in CD4+ T cells and avoid innate antiviral immune responses in HIV-1 infected individuals on ART. To test this hypothesis and to facilitate the development of a novel strategy for HIV-1 cure, we designed three specific aims in two phases: (1) R61 phase (years 1-3): Aim 1. To determine m6A profile of HIV-1 RNA in subsets of CD4+ T cells from ART- treated patients; Aim 2. To identify cellular targets in the m6A pathway important for HIV-1 reactivation in primary CD4+ T cells; and (2) R33 phase (years 4-5): Aim 3. To examine anti-HIV-1 effects of small molecules inhibiting m6A modifications in primary CD4+ T cells. Overall Impact: These studies will reveal how m6A modifications of HIV-1 RNA regulate viral latency in ART- treated patients. The studies in the R61 phase will define new mechanisms of HIV-1 persistence and identify potential therapeutic targets. The R33 phase study will develop an m6A-specific strategy to inhibit HIV-1 replication in primary CD4+ T cells.

项目概要/摘要尽管抗逆转录病毒疗法 (ART) 非常有效，但静息 CD4+ T 细胞中的潜在 HIV-1 储存库是 HIV-1 感染功能性治愈的主要障碍。我们这个 R61/R33 双相项目的总体目标是：了解 HIV-1 感染者中 HIV-1 RNA 转录后调控的机制正在进行 ART（探索性 R61 阶段），并开发一种新的治疗策略来改变 RNA 后转录修饰作为抑制 HIV-1 复制的潜在治疗平台（Developmental R33相）。我们的多学科小组有独特的能力来解决本次资助中强调的几个关键问题机会。我的实验室是独立发现 N6-甲基腺苷 (m6A) 的三个小组之一 HIV-1 RNA 的修饰可在体外调节 CD4+ T 细胞中的病毒复制。使用 CD4+ T 细胞系和来自健康捐赠者的原代 CD4+ T 细胞，我们研究了 m6A 修饰的机制调节 HIV-1 感染。我们还发现 HIV-1 RNA 的 m6A 修饰抑制先天抗病毒免疫来自健康捐赠者的原代巨噬细胞的反应。我们的体外研究表明 m6A 修饰 HIV-1 RNA 在病毒复制和对病毒感染的先天免疫反应中发挥着关键作用。然而， HIV-1 RNA m6A 修饰在调节 HIV-1 感染者 ART 病毒复制中的作用仍然未知。我们的目标是填补这一重要的知识空白，并将研究结果转化为潜在的反 HIV-1 疗法。我们假设 HIV-1 RNA 的 m6A 修饰有助于在 CD4+ T 细胞中建立和维持病毒潜伏期并避免 HIV-1 感染者在接受 ART 时出现先天性抗病毒免疫反应。为了检验这个假设并为了促进 HIV-1 治疗新策略的制定，我们分两个阶段设计了三个具体目标： (1) R61 期（第 1-3 年）：目标 1. 确定来自 ART- 的 CD4+ T 细胞亚群中 HIV-1 RNA 的 m6A 谱接受治疗的患者；目标 2. 确定 m6A 通路中对原发性 HIV-1 重新激活很重要的细胞靶点 CD4+T细胞； (2) R33 阶段（第 4-5 年）：目标 3. 检查小分子抑制的抗 HIV-1 效果原代 CD4+ T 细胞中的 m6A 修饰。总体影响：这些研究将揭示 HIV-1 RNA 的 m6A 修饰如何调节 ART 中的病毒潜伏期接受治疗的患者。 R61 阶段的研究将定义 HIV-1 持续存在的新机制并确定潜在的治疗靶点。 R33期研究将开发一种m6A特异性策略来抑制HIV-1 原代 CD4+ T 细胞中的复制。