N6-Methyladenosine Methylome in Duchenne Muscular Dystrophy

N6-甲基腺苷甲基化在杜氏肌营养不良症中的应用

基本信息

批准号：
10593310
负责人：
Bijan K Dey
金额：
$ 17.2万
依托单位：
STATE UNIVERSITY OF NEW YORK AT ALBANY
依托单位国家：
美国
项目类别：
财政年份：
2023
资助国家：
美国
起止时间：
2023-03-06 至 2026-02-28
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10593310
关键词：
Affect Age Alkylation Area Binding Proteins Biological Assay Biological Models Biological Process Biology Cell Count Cessation of life Chemicals Child Complex Data Degenerative Disorder Development Disease Duchenne muscular dystrophy Dystrophin Enzymes Foundations Functional disorder Future Gene Expression Regulation Genes Genetic Half-Life Homologous Gene IGF2 gene Immunoprecipitation Individual Injections Knowledge Link Longevity Maps Mediating Messenger RNA Methods Methylation Methyltransferase Modeling Modification Molecular Morphology Mus Muscle Muscle Fibers Muscle satellite cell Muscular Dystrophies Mutation Myoblasts Natural regeneration Nucleotides Pathogenesis Pathway interactions Patients Phenotype Play Post-Transcriptional Regulation Process Production Proteins Quantitative Reverse Transcriptase PCR RNA RNA Splicing RNA Stability RNA immunoprecipitation sequencing Reader Research Resolution Role Skeletal Muscle Solid Symptoms Techniques Testing Translations Wheelchairs boys clinically relevant crosslink early childhood effective therapy epitranscriptomics experimental study fat mass and obesity-associated protein human disease improved inhibitor innovation insight knock-down mRNA Stability mdx mouse methylome mouse model muscle degeneration muscle regeneration myogenesis novel premature repaired skeletal muscle wasting therapeutic target therapy development transcriptome

项目摘要

PROJECT SUMMARY/ABSTRACT Duchenne muscular dystrophy (DMD) is a fatal X-linked muscle degenerative disease. Children with DMD become restricted to wheelchairs within the first decade of their lives and die within the third. There is no effective treatment available for this devastating disease. Key myogenic processes in DMD pathophysiology at the molecular level are not completely understood, hindering the development of therapies for DMD. To fill this knowledge gap, we have been investigating whether an emerging mode of post-transcriptional gene regulation, known as epitranscriptomics, plays a role in DMD pathogenesis. Epitranscriptomics is a dynamic process that regulates RNA stability, splicing, and translation by reversible chemical modifications of mRNAs. N6- methyladenosine (m6A) is the most abundant epitranscriptomic mark. The m6A marks are installed by methyltransferase like 3 (METTL3) and methyltransferase like 14 (METTL14) and erased by fat mass and obesity-associated (FTO) and alkylation repair homolog 5 (ALKBH5). Specific reader proteins selectively recognize m6A-modified mRNAs and control their fates. Our recent findings revealed that m6A mRNA methylation regulates myogenesis in DMD and that levels of the m6A writers, erasers, and m6A marks are tightly regulated during myoblast differentiation, muscle regeneration, and in DMD. These studies suggest that m6A mRNA methylation plays a pivotal role in DMD pathophysiology. Here, we propose to gain a deeper mechanistic insight into m6A mRNA methylation in DMD. We will perform a transcriptome-wide analysis to map m6A-modified mRNAs (methylome) in skeletal muscle stem cells (MuSCs), primary myoblasts, and skeletal muscle of a well- established mdx mouse model of DMD and its counterpart healthy mice. We will also map the m6A methylome in DMD patient-derived and healthy myoblasts to extend our conclusions to a more clinically relevant model system. We will mechanistically characterize a select subset of prioritized m6A target mRNAs, both previously associated with DMD, as well as the novel, to establish their link to muscle degeneration in DMD. Our proposed research will make a significant impact by revealing transcriptome-wide m6A target mRNAs in DMD and provide insights into DMD pathophysiology incurred by the dysregulation of this novel mechanism.

项目概要/摘要杜氏肌营养不良症 (DMD) 是一种致命的 X 连锁肌肉退行性疾病。患有DMD的儿童在生命的前十年内只能使用轮椅，并在第三个十年内死亡。没有有效的针对这种毁灭性疾病的治疗方法。 DMD 病理生理学中的关键生肌过程分子水平尚未完全了解，阻碍了 DMD 疗法的开发。为了填补这个知识差距，我们一直在研究是否有一种新兴的转录后基因调控模式，称为表观转录组学，在 DMD 发病机制中发挥作用。表观转录组学是一个动态过程通过 mRNA 的可逆化学修饰来调节 RNA 稳定性、剪接和翻译。 N6- 甲基腺苷 (m6A) 是最丰富的表观转录组标记。 m6A 标记由以下人员安装甲基转移酶样 3 (METTL3) 和甲基转移酶样 14 (METTL14) 并被脂肪量消除肥胖相关 (FTO) 和烷基化修复同源物 5 (ALKBH5)。选择性地特定读取蛋白识别 m6A 修饰的 mRNA 并控制它们的命运。我们最近的研究结果表明 m6A mRNA 甲基化调节 DMD 中的肌生成，并且 m6A 写入器、擦除器和 m6A 标记的水平受到严格调节在成肌细胞分化、肌肉再生和 DMD 过程中。这些研究表明 m6A mRNA 甲基化在 DMD 病理生理学中起着关键作用。在这里，我们建议获得更深入的机制洞察 DMD 中 m6A mRNA 甲基化。我们将进行全转录组分析以绘制 m6A 修饰图谱骨骼肌干细胞 (MuSC)、原代成肌细胞和良好骨骼肌中的 mRNA（甲基化组）建立DMD mdx小鼠模型及其对应的健康小鼠。我们还将绘制 m6A 甲基化组图在 DMD 患者来源的和健康的成肌细胞中，将我们的结论扩展到更具临床相关性的模型系统。我们将机械地表征优先 m6A 目标 mRNA 的选定子集，这两个都是之前的与 DMD 以及小说相关，以确定它们与 DMD 肌肉退化的联系。我们提出的研究将通过揭示 DMD 中转录组范围的 m6A 靶 mRNA 产生重大影响，并提供深入了解这种新机制失调引起的 DMD 病理生理学。