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

• 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
• 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

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.

项目摘要/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。DMD儿童 在他们生命的头十年里，他们只能坐轮椅，在第三个十年内死亡。没有有效 治疗这种毁灭性的疾病。DMD病理生理学中的关键生肌过程 分子水平的研究还不完全清楚，阻碍了DMD治疗方法的发展。填补这一 知识差距，我们一直在研究是否一种新兴的转录后基因调控模式， 称为表转录组学，在DMD发病机制中起作用。表位转录组学是一个动态过程， 通过对mRNA进行可逆的化学修饰来调节RNA的稳定性、剪接和翻译。N6- 甲基腺苷（m6 A）是最丰富的表转录组标记。m6 A标志由 甲基转移酶样3（胃L3）和甲基转移酶样14（胃L14），并通过脂肪量消除， 肥胖相关（FTO）和烷基化修复同源物5（ALKBH 5）。特异性阅读蛋白 识别m6 A修饰的mRNA并控制它们的命运。我们最近的研究结果表明，m6 A mRNA甲基化 调节DMD中的肌生成，并且m6 A写入器、擦除器和m6 A标记的水平受到严格调节 在成肌细胞分化、肌肉再生和DMD中。这些研究表明，m6 A mRNA 甲基化在DMD病理生理学中起关键作用。在这里，我们建议获得更深层次的机械洞察力， m6 A mRNA甲基化。我们将进行全转录组分析以绘制m6 A修饰的图谱 骨骼肌干细胞（MuSC）、原代成肌细胞和骨骼肌中的mRNA（甲基化组） 建立了mdx小鼠DMD模型及相应的健康小鼠模型。我们还将绘制m6 A甲基化组， 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临床相关的模型 系统我们将机械地表征一个选择的优先m6 A靶mRNA的子集， 与DMD相关，以及新的，以建立他们的联系，肌肉变性的DMD。我们提出的 研究将通过揭示DMD中转录组范围的m6 A靶mRNA产生重大影响，并提供 深入了解DMD病理生理学所引起的失调，这种新的机制。