The role of m6A RNA modification as modulator of dsRNA induced cell-intrinsic innate immune responses in hematopoiesis

m6A RNA 修饰作为 dsRNA 调节剂在造血过程中诱导细胞内在先天免疫反应的作用

基本信息

批准号：
10152825
负责人：
Stephanie Halene
金额：
$ 29.48万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2021
资助国家：
美国
起止时间：
2021-08-01 至 2024-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10152825
关键词：
Adult Affect Antiviral Agents Applications Grants Biological Assay Catalytic Domain Cells Complex Data Defect Double-Stranded RNA Dysmyelopoietic Syndromes Engraftment Failure Fetal Development Fetal Liver Generations Genetic Genetic Transcription Hematopoiesis Hematopoietic Hematopoietic stem cells Immune Response Genes Immune response Immune signaling Immunoprecipitation In Vitro Innate Immune Response Interferons Knock-out Knockout Mice Light Maps Mediating Messenger RNA Methyltransferase Modeling Modification Nucleotides Pathway interactions Phenotype Play Process Proteins RNA RNA metabolism Reader Reporting Resolution Ribonucleases Role Signal Transduction Structure Transcript Transcription Initiation Site Translations Transplantation Up-Regulation Work epitranscriptome functional disability in vivo innate immune pathways knock-down leukemia mouse model novel oligoadenylate pathogen preservation prevent progenitor response sensor single-cell RNA sequencing stem cell function

项目摘要

Project Summary N6-methyladenosine (m6A) is the most abundant RNA modification and plays key roles in RNA metabolism. m6A writers and readers are highly expressed in hematopoietic stem and progenitor cells and dysregulated in myelodysplasia and leukemia. We show that loss of Mettl3 in hematopoietic stem and progenitor cells (HSPCs) resulted in upregulation of an anti-viral innate immune response signature as well as marked proliferation and differentiation defects. In particular, we are the first to report that loss of m6A RNA modification resulted in aberrant dsRNA formation, leading to activation of the MDA5-RIG-I, PKR-eIF2A, and OAS-RNAse L pathways. By differential immunoprecipitation of dsRNAs from Mettl3 KO versus WT cells, we identified a specific subset of long, normally highly m6A modified transcripts with low folding energies, signifying propensity to form extensive RNA secondary structures. Disruption of innate immune signaling via deletion of Mavs or knockdown of Rnasel in part rescued colony formation in vitro and engraftment in competitive transplantation assays in vivo. These data suggest that one of the roles of m6A RNA modifications is to maintain single-strandedness of endogenous RNAs whereby they mark RNAs as “self” and distinguish them from exogenous pathogen-derived dsRNAs. In this SHINE II application we propose to dissect the role of m6A RNA modification in HSPCs specifically as it pertains to its role as suppressor of aberrant innate immune signaling. We will determine the mechanism by which METTL3-mediated m6A RNA modification prevents abnormal dsRNA formation and consequent induction of interferon signaling, we will dissect how m6A loss results in a deleterious immune response that disrupts HSPC function, and we will assess the universality this phenomenon across hematopoietic lineages.

项目摘要 N6-甲基腺苷（M6A）是最丰富的RNA修饰，在RNA代谢中起关键作用。 M6A 作家和读者在造血茎和祖细胞中高度表达，在骨髓增生和白血病。我们显示，造血茎和祖细胞（HSPC）中Mettl3的损失导致上调抗病毒先天免疫响应特征以及明显的增殖和分化缺陷。特别是，我们是第一个报告M6A RNA修饰的损失导致异常DSRNA形成，导致MDA5-RIG-I，PKR-EIF2A和OAS-RNase L途径的激活。通过差异从Mettl3 KO与WT细胞的DSRNA免疫沉淀，我们确定了长长的特定子集，通常是高度M6A修饰的成绩单低折叠能，表示形成广泛的RNA次级的承诺结构。通过删除MAV或敲除RNASEL的一部分被救出，对先天免疫信号的破坏体外的体外形成和植入体内的竞争移植测定法。这些数据表明 M6a RNA修饰的作用之一是维持内源性RNA的单链性它们将RNA标记为“自我”，并将其与外源性病原体衍生的DSRNA区分开。在此Shine II中，我们建议在HSPC中剖析M6A RNA修饰的作用与其作为异常先天免疫信号传导的抑制剂有关。我们将通过 Mettl3介导的M6A RNA修饰可防止异常的DSRNA形成，因此诱导在干扰信号传导中，我们将剖析M6A损失如何导致有害免疫反应破坏 HSPC功能，我们将评估跨造血谱系的宇宙。