Preclinical models, biomarkers, and therapy for myotonic dystrophy type 1

1 型强直性肌营养不良的临床前模型、生物标志物和治疗

基本信息

批准号：
10237267
负责人：
MAURICE SCOTT SWANSON
金额：
$ 49.21万
依托单位：
UNIVERSITY OF ROCHESTER
依托单位国家：
美国
项目类别：
财政年份：
2003
资助国家：
美国
起止时间：
2003-09-30 至 2023-08-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10237267
关键词：
3&apos Untranslated Regions Address Adult Affect Alleles Animal Model Antisense Oligonucleotides Binding Biological Markers C9ORF72 CRISPR/Cas technology CUG repeat Coupled Cyclin-Dependent Kinases DNA Polymerase II DNA-Directed RNA Polymerase Defect Development Disease Drug Approval Event Exercise Exons Experimental Models Family Fragile X Syndrome Generations Genes Genetic Transcription Genome Goals Health Hereditary Disease Hexoses Histopathology Human In Vitro Insulin Knock-in Knock-in Mouse Length Ligands Link Mediating Messenger RNA Methods MicroRNAs Microsatellite Repeats Modeling Modification Molecular Mus Muscle Muscular Dystrophies Mutation Myocardium Myotonia Myotonic Dystrophy Myotonic dystrophy type 1 Neuromuscular Diseases Outcome Measure Pathogenesis Pathogenicity Pathway interactions Patient observation Patients Pharmaceutical Preparations Pharmacodynamics Phosphotransferases Pre-Clinical Model Program Development Proteins RNA RNA Processing RNA Splicing RNA-Binding Proteins Recovery of Function Regulation Research Residual state Safety Series Skeletal Muscle Smooth Muscle Testing Therapeutic Therapeutic Intervention Toxic effect Transcription Elongation Translations Tremor/Ataxia Syndrome Untranslated RNA analog base biomarker development combinatorial design drug development effective therapy frontotemporal lobar dementia-amyotrophic lateral sclerosis functional disability improved in vivo inhibitor/antagonist knock-down loss of function mRNA Precursor mouse model muscular structure mutant novel preclinical study repaired small molecule targeted treatment therapeutic development therapeutic target therapeutically effective therapy development tool transcriptome transcriptome sequencing treatment response

项目摘要

Myotonic dystrophy type 1 (DM1), which is caused by CTG expansions (CTGexp) in the 3' untranslated region of the DMPK gene, has been used as a model for RNA-mediated disease mechanisms associated with other microsatellite expansion diseases, including fragile X tremor/ataxia syndrome (FXTAS) and C9orf72 amyotrophic lateral sclerosis and frontotemporal dementia (C9-ALS/FTD). In DM1, transcription of the CTGexp mutation results in CUGexp RNAs that alter the developmental regulation of pre-mRNA processing and mRNA localization events mediated by the MBNL and CELF families of RNA binding proteins. However, additional cellular pathways, such as miRNA processing and repeat-associated non-AUG translation, have also been implicated in DM1 pathogenesis. Most importantly, no effective therapies exist to treat this neuromuscular disease. To address these deficiencies, this project is designed to generate more informative mouse experimental models for DM1 to elucidate the relative contribution of each of the proposed pathomechanisms and qualify RNA splicing defects as responsive biomarkers of therapeutic response with the goal of developing effective therapeutic approaches to decrease the toxic burden of CUGexp RNAs. Aim 1 builds upon our recent development of Dmpk CTGexp knockin mice generated using a combination of rolling circle amplification to generate large repeats in vitro and CRISPR/Cas9-mediated genome modification. Using an allelic series of increasing CTG repeat lengths that represent the late-onset to congenital spectrum of the DM1 pathogenic range, we will determine CTG length-dependent effects on skleletal and heart muscle structure/function, RNA processing/localization/turnover and RAN translation. Transcriptome analysis will be pursued further in Aim 2, which is based upon our prior observations that patient functional impairment corresponds to RNA splicing defects and MBNL loss of function, to determine if splicing defects qualify as effective biomarkers that are responsive to CUGexp levels, MBNL activity and therapeutic intervention. In Aim 3, we will broaden this therapeutic scope and evaluate multiple strategies, including antisense oligonucleotide (ASO)-mediated CUGexp knockdown and small molecule approaches to inhibit transcription of mutant Dmpk CTGexp genes. The overall objective of this project is to provide the DM field with more robust mouse models of DM1 while also evaluating splicing defects as biomarkers of disease status and developing single small molecule strategies

强直性肌营养不良 1 型 (DM1)，由 3' 非翻译区的 CTG 扩张 (CTGexp) 引起 DMPK 基因的 DMPK 基因已被用作与其他疾病相关的 RNA 介导的疾病机制的模型微卫星扩张疾病，包括脆性 X 震颤/共济失调综合征 (FXTAS) 和 C9orf72 肌萎缩侧索硬化症和额颞叶痴呆 (C9-ALS/FTD)。在 DM1 中，转录 CTGexp 突变导致 CUGexp RNA 改变前 mRNA 加工的发育调节以及由 RNA 结合蛋白的 MBNL 和 CELF 家族介导的 mRNA 定位事件。然而，其他细胞途径，例如 miRNA 加工和重复相关的非 AUG 翻译，也已与 DM1 发病机制有关。最重要的是，目前还没有有效的疗法来治疗这种情况神经肌肉疾病。为了解决这些缺陷，该项目旨在生成更多信息 DM1 的小鼠实验模型，以阐明每个提议的相对贡献病理机制并将 RNA 剪接缺陷鉴定为治疗反应的反应性生物标志物开发有效的治疗方法以减少 CUGexp RNA 的毒性负担的目标。目标 1 构建我们最近开发了使用滚环组合产生的 Dmpk CTGexp 敲入小鼠体外扩增以产生大量重复序列以及 CRISPR/Cas9 介导的基因组修饰。使用 CTG 重复长度增加的等位基因系列，代表 DM1 的晚发型到先天性谱系致病范围，我们将确定 CTG 长度依赖性对骨骼和心肌的影响结构/功能、RNA 加工/定位/周转和 RAN 翻译。转录组分析将在目标 2 中进一步追求，这是基于我们之前的观察，即患者功能障碍对应于 RNA 剪接缺陷和 MBNL 功能丧失，以确定剪接缺陷是否符合对 CUGexp 水平、MBNL 活性和治疗干预有反应的有效生物标志物。瞄准 3、我们将扩大这种治疗范围并评估多种策略，包括反义寡核苷酸 (ASO) 介导的 CUGexp 敲低和小分子方法抑制突变 Dmpk 的转录 CTGexp 基因。该项目的总体目标是为DM领域提供更强大的小鼠模型 DM1 的同时还评估剪接缺陷作为疾病状态的生物标志物并开发单个小分子策略