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.

项目摘要/摘要尽管高效的抗逆转录病毒疗法（ART），静止的CD4+ T细胞中的潜在HIV-1储存剂是 HIV-1感染功能治愈的主要障碍。我们这个R61/R33双重项目的总体目标是：了解HIV-1感染者HIV-1 RNA转录后调节的机制正在使用ART（探索性R61阶段），并制定一种新的治疗策略，以改变后RNA 转录修饰是抑制HIV-1复制的潜在治疗平台（发展 R33阶段）。我们的多学科小组有独特的准备解决此资金中突出显示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）的三组之一 HIV-1 RNA的修饰在体外调节CD4+ T细胞中的病毒复制。使用CD4+ T细胞系和来自健康供体的主要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助者的主要巨噬细胞的反应。我们的体外研究表明M6A修改 HIV-1 RNA在病毒复制和对病毒感染的先天免疫反应中起关键作用。但是， HIV-1 RNA修饰在调节HIV-1感染者在ART中调节病毒复制中的作用仍然未知。我们旨在填补这一重要的知识差距，并将发现转化为潜在的反 - HIV-1治疗学。我们假设HIV-1 RNA的M6A修饰有助于建立和维持CD4+ T细胞中的病毒潜伏期并避免在HIV-1感染的ART上感染的HIV-1感染者中的先天抗病毒免疫反应。检验这一假设和为了促进HIV-1治疗的新型策略的发展，我们在两个阶段设计了三个特定目标：（1）R61阶段（1-3年）：目标1。确定来自Art-的CD4+ T细胞子集中HIV-1 RNA的M6A谱。治疗的患者； AIM 2。确定M6A途径中对HIV-1重新激活至关重要的细胞靶标 CD4+ T细胞；（2）R33相（4-5年）：目标3。检查抑制小分子的抗HIV-1效应主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修改如何调节艺术中的病毒潜伏期治疗的患者。 R61阶段的研究将定义HIV-1持久性的新机制，并确定潜在的治疗靶标。 R33阶段研究将制定M6A特异性策略来抑制HIV-1 一级CD4+ T细胞中的复制。