HIV-1-induced upregulation of m6A modifications of cellular RNA in CD4+ T-cells

项目摘要/摘要 这种R21应用是对PA-19-237的响应：病毒学(包括艾滋病毒)和免疫中的新RNA 法规：基础科学和治疗发现。在这个新的项目中，我们建议研究 HIV-1诱导细胞RNA N6-甲基腺苷(M6A)修饰上调及其机制 原代CD4+T细胞。通过跨学科协作和技术创新，我们的团队是独一无二的 准备解决此次融资机会中强调的关键问题。 我们研究了m6A修饰调节HIV-1感染原代CD4+T细胞的机制 细胞。有趣的是，我们发现HIV-1包膜蛋白gp120上调了细胞rna的m6A水平。 与病毒在原代CD4+T细胞中的复制无关。我们进一步的研究表明这种相互作用 Gp120和CD4受体之间的关系在HIV-1诱导的M6A上调CD4+细胞RNA中起重要作用 T细胞。然而，HIV-1gp120诱导的功能意义和分子机制 CD4+T细胞中细胞RNA的m6A修饰上调仍不清楚。我们的目标是填补这些重要的 拟议研究中的知识差距也与供资机会的目标很好地一致。 我们假设HIV-1gp120介导的m6A修饰在初级CD4+细胞中上调细胞RNA T细胞通过改变表位转录的m6A谱和细胞基因表达来增强病毒复制。我们 设计了两个特定的目标来检验中心假设：目标1.外转录和转录分析 HIV-1感染或gp120处理的CD4+T细胞；以及目的2.探讨gp120诱导m6A的机制 上调及其对原代CD4+T细胞活化和HIV-1复制的影响。 整体影响。定义CD4+T细胞的m6A表位转录和一般转录图谱将揭示 参与调控HIV-1感染的细胞RNA。我们的探索性研究可以确定新的宿主因素 对于调节HIV-1复制和持久性至关重要。预期的结果将有助于定义功能 HIV-1gp120诱导原代CD4+T细胞RNA m6A水平上调的意义 为HIV-1在CD4+T细胞中持续存在的分子机制提供新的见解。

Project Summary/Abstract This R21 application is in response to PA-19-237: Novel RNAs in Virology (including HIV) and Immune Regulation: Basic Science and Therapeutic Discovery. In this new project, we propose to investigate the function and mechanisms of HIV-1-induced upregulation of N6-methyladenosine (m6A) modifications of cellular RNA in primary CD4+ T cells. Through interdisciplinary collaborations and technical innovations, our team is uniquely poised to address key questions highlighted in this funding opportunity. We have investigated the mechanisms by which m6A modifications regulate HIV-1 infection of primary CD4+ T cells. Interestingly, we found that the HIV-1 envelope protein gp120 upregulates m6A levels of cellular RNA independently of viral replication in primary CD4+ T cells. Our further study suggested that the interaction between gp120 and the CD4 receptor is important for HIV-1-induced m6A upregulation of cellular RNA in CD4+ T cells. However, the functional significance and the molecular mechanisms of HIV-1 gp120-induced upregulation of m6A modifications of cellular RNA in CD4+ T cells remain unknown. We aim to fill these significant knowledge gaps through the proposed studies that are also aligned well with the goal of the funding opportunity. We hypothesize that HIV-1 gp120-mediated upregulation of m6A modifications of cellular RNA in primary CD4+ T cells enhances viral replication by altering the epitranscriptomic m6A profile and cellular gene expression. We designed two specific aims to test the central hypothesis: Aim 1. Epitranscriptomic and transcriptomic analyses of HIV-1-infected or gp120-treated CD4+ T cells; and Aim 2. Investigate the mechanisms of gp120-induced m6A upregulation and the effects on primary CD4+ T cell activation and HIV-1 replication. Overall impact. Defining m6A epitranscriptomic and general transcriptomic profiles of CD4+ T cells will reveal cellular RNAs involved in regulating HIV-1 infection. Our exploratory studies can identify novel host factors that are critical for modulating HIV-1 replication and persistence. The expected results will help define the functional significance of HIV-1 gp120-induced upregulation of cellular RNA m6A levels in primary CD4+ T cells, which will provide new insights into the molecular mechanisms of HIV-1 persistence in CD4+ T cells.