Identification of components and mechanisms regulating expanded CUG-repeat RNP complexes in Myotonic Dystrophy Type 1 muscle cells

强直性肌营养不良 1 型肌细胞中调节扩展 CUG 重复 RNP 复合物的成分和机制的鉴定

• 批准号: 10667708
• 负责人: Thomas A Cooper
• 金额: $ 21.12万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-01 至 2025-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10667708
• 关键词: 3' Untranslated Regions; Accounting; Address; Adult; Affect; Alleles; Applications Grants; Binding; Binding Proteins; Biotinylation; CUG repeat; Cell Line; Cell Nucleus; Cells; Chimeric Proteins; Complex; Equilibrium; Family; Fluorescent in Situ Hybridization; Foundations; Genes; Genetic Transcription; Goals; Guide RNA; Hereditary Disease; Homeostasis; Human; Image; Individual; Investigation; Knowledge; Label; Link; Mass Spectrum Analysis; Mediating; Messenger RNA; Molecular; Molecular and Cellular Biology; Muscle Cells; Muscular Atrophy; Muscular Dystrophies; Myoblasts; Myotonic Dystrophy; Myotonic dystrophy type 1; Nature; Nuclear; Nuclear Structure; Pathogenesis; Pathogenicity; Pathologic; Physiological; Proteins; RNA; RNA Folding; RNA Processing; RNA-Binding Proteins; Ribonucleoproteins; Role; Signal Pathway; Signal Transduction; Skeletal Muscle; Structure; Tetanus Helper Peptide; Therapeutic; Tissue Sample; Tissues; Toxic effect; United States; United States National Institutes of Health; Validation; gain of function; insight; knock-down; loss of function; mortality; mouse model; mutant; new therapeutic target; novel; paralogous gene; skeletal muscle weakness; therapeutic target; tissue fixing

Project Summary Myotonic dystrophy (DM) is the most common cause of adult-onset muscular dystrophy and the second most common cause of muscular dystrophy overall. Myotonic dystrophy type 1 (DM1) results from an expanded CTG repeat in the 3’ untranslated region of the DMPK gene. The molecular basis for pathogenesis is a toxic gain of function of the RNA transcribed from the mutant allele that contains long tracts of expanded CUG repeats (CUGexp RNA). CUGexp RNA remains in the nucleus bound with proteins to form ribonucleoprotein complexes (RNPs) detected as foci by RNA fluorescence in situ hybridization. CUGexp RNPs include the Muscleblind-Like (MBNL) paralogs, MBNL1 and MBNL2, that are sequestered resulting in their loss of function and a primary cause of pathogenesis. CUGexp RNPs are dynamic nuclear structures that are the cause and therapeutic target of DM1 pathogenesis yet knowledge of the composition of the CUGexp RNP is limited and a full accounting of the mechanisms of CUGexp RNA toxicity in skeletal muscle remains to be established. To gain insight into the molecular and cellular biology of CUGexp RNA and the CUGexp RNP in skeletal muscle, we will use proximity labeling to identify protein and RNA components of the CUGexp RNP complex in addition to MBNL and CUGexp RNA and determine their roles in CUGexp RNA pathogenesis in skeletal muscle. The results will provide the foundation to identify novel therapeutic targets and enhance the efficiency of current approaches targeting the CUGexp RNA component of the RNP.

项目摘要 肌营养不良症（DM）是成人发作肌营养不良的最常见原因，第二个是最常见的 总体上肌营养不良的常见原因。肌发育症1型（DM1）来自扩展 CTG在DMPK基因的3'未翻译区域中重复。发病机理的分子基础是有毒 从突变等位基因转录的RNA的功能获得，其中包含长长的cug群 重复（CugeXP RNA）。 CugeXP RNA保留在与蛋白质结合的细胞核中，形成核糖核蛋白 通过RNA荧光原位杂交检测为焦点的复合物（RNP）。 cugexp rnps包括 肌肉闪光（MBNL）旁系同源物MBNL1和MBNL2，它们被隔离，导致其功能丧失 以及发病机理的主要原因。 CugeXP RNP是动态核结构，是原因和 DM1发病机理的治疗靶点却了解CugeXP RNP的组成量是有限的，并且 尚待确定CugeXP RNA毒性机制的全部记录尚待确定。到 深入了解CugeXP RNA的分子和细胞生物学和骨骼肌中的CugeXP RNP， 我们将使用接近标记来鉴定CugeXP RNP复合物的蛋白质和RNA成分。 到MBNL和CugeXP RNA，并确定它们在骨骼肌中CugeXP RNA发病机理中的作用。这 结果将为识别新型热目标并提高电流的效率提供基础 接近针对RNP的CugeXP RNA成分的接近。