RNA processing-mediated mechanisms of CNS dysfunction in Myotonic Dystrophy

RNA加工介导强直性肌营养不良中枢神经系统功能障碍的机制

基本信息

批准号：
10055974
负责人：
GARY J BASSELL
金额：
$ 2.63万
依托单位：
UNIVERSITY OF FLORIDA
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-07-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10055974
关键词：
3&apos Untranslated Regions Adhesives Adverse event Affect Anesthesia procedures Anesthetics Animal Model Antisense Oligonucleotides Attenuated Autopsy Award Behavior Behavioral Behavioral Research Benzodiazepines Biological Assay Blood Vessels Brain Cessation of life Chemosensitization Clinical Data Defect Dexmedetomidine Diagnosis Diazepam Disease Dominant Genetic Conditions Event Exhibits Functional disorder Future Gene Expression Profiling General Anesthesia General anesthetic drugs Genes Genetic Diseases Goals Heart Homeostasis Human Impairment Isoflurane Knock-out Knockout Mice Laboratories Lead Link Mediating Modeling Molecular Mus Myotonic Dystrophy N-Methylaspartate NMDA receptor A1 National Research Service Awards Neuraxis Neurons New York Operative Surgical Procedures Outcome Parents Patients Perioperative Phenotype Physiological Play Population Post-Transcriptional Regulation Prevalence Procedures Protein Isoforms Protein Kinase Proteins Public Health RNA RNA Processing RNA Splicing RNA-Binding Proteins Reaction Receptors, Adrenergic, alpha-2 Recovery Research Role Scientist Sedation procedure Skeletal Muscle Smooth Muscle Structure Symptoms Synapses Techniques Testing Therapeutic Tissues Travel Universities Variant Vertebrates Viral Work base behavioral phenotyping brain dysfunction brain tissue clinical phenotype experience gamma-Aminobutyric Acid genome-wide improved insight mouse model neuromuscular neurotransmission novel therapeutics outcome prediction overexpression receptor respiratory response restoration sedative sevoflurane training project transcriptome transcriptome sequencing transmission process

项目摘要

Myotonic dystrophy (dystrophia myotonica, DM) is an autosomal dominant genetic disease, with a diagnosed prevalence of 1:8000 people worldwide, that affects multiple tissues of the body, including skeletal muscle, heart, and related to this proposal, the central nervous system (CNS). DM1 is caused by expanded CTG repeats in the 3' UTR of dystrophia myotonica protein kinase (DMPK). There is substantial evidence in mouse DM1 models and human DM1 postmortem tissue to support an RNA-mediated disease mechanism where toxic intranuclear CUG RNA foci sequester Muscleblind (MBNL) RNA binding proteins that normally play crucial roles to regulate various aspects of post-transcriptional gene regulation. A major gap in our understanding is that we do not know which RNA processing defects underlie specific impairments in DM1 brain function. Recent work together with our new findings suggests that missplicing of RNAs encoding synaptic proteins is responsible for CNS dysfunction in DM1. Our central hypothesis is that CNS phenotypes are directly attributed to loss of MBNL mediated RNA processing and that restoration of MBNL activity and/or splicing can restore brain function. Our goal is to gain a thorough understanding of RNA processing-mediated mechanisms of CNS dysfunction in DM1 and use this to develop and rigorously evaluate novel therapeutic strategies. The overall objectives of this proposal are to use both candidate and genome wide approaches, applied to MBNL KO mice and a new AAV9 based neuronal mouse model, compared to RNAseq analysis of human postmortem brain, to evaluate the role of specific splicing events to drive symptoms, and to comprehensively identify changes in missplicing and RNA processing. Aim 1 will characterize how dysregulation of GABRG2, GRIN1, and SNAP25 splicing events is linked to molecular, cellular, and behavioral phenotypes observed in DM1. Aim 2 will develop a new AAV9 based mouse model to elucidate the set of RNA processing events in neurons that cause DM1 phenotypes, through transcriptional profiling and overlap of human DM brains with DM mouse model brains. Aim 3 will assess the extent to which antisense oligonucleotides or MBNL expression can rescue molecular, cellular, physiologic and behavioral phenotypes in DM1 mouse models. These studies will provide new mechanistic insights into how perturbations to specific RNA processing events can lead to CNS symptoms in myotonic dystrophy, and provide a broader comprehensive view of all transcriptome changes occurring in the DM CNS. The proposed work is significant, as no molecular changes have been linked to any phenotypes in the DM CNS. This provides the framework for future therapeutic efforts aimed at correcting CNS defects.

肌营养不良症（DM肌营养不良，DM）是一种常染色体显性遗传疾病，已诊断为全球1：8000人的患病率会影响身体的多个组织，包括骨骼肌，心脏，与该提议有关，中枢神经系统（CNS）。 DM1是由扩展的CTG引起的在肌营养不良蛋白蛋白激酶（DMPK）的3'UTR中重复。小鼠有大量证据 DM1模型和人DM1验尸组织支持RNA介导的疾病机制，其中有毒核内CUG RNA焦点次肌肉蓝线（MBNL）RNA结合蛋白，通常播放调节转录后基因调控的各个方面的关键作用。我们的主要差距理解是，我们不知道DM1中哪些RNA处理缺陷是特定损伤的基础大脑功能。最近的工作以及我们的新发现表明，错过了RNA编码突触蛋白负责DM1中的CNS功能障碍。我们的中心假设是CNS表型直接归因于MBNL介导的RNA处理的丢失以及MBNL活性的恢复和/或剪接可以恢复大脑功能。我们的目标是对RNA处理介导的透彻了解 DM1中CNS功能障碍的机制，并使用它来开发和严格评估新型治疗策略。该提案的总体目标是同时使用候选和基因组范围的方法，与RNASEQ分析相比人类验尸大脑，评估特定剪接事件以驱动症状的作用，并全面确定错过和RNA处理的变化。 AIM 1将表征如何 GABRG2，GRIN1和SNAP25剪接事件的失调与分子，细胞和行为有关在DM1中观察到的表型。 AIM 2将开发新的基于AAV9的鼠标模型，以阐明RNA集通过转录分析和重叠的神经元的处理事件引起DM1表型 DM小鼠模型大脑的人DM大脑。 AIM 3将评估反义的程度寡核苷酸或MBNL表达可以挽救分子，细胞，生理和行为表型 DM1鼠标模型。这些研究将提供有关如何扰动特定的新机械见解 RNA处理事件可以导致Myotonic营养不良的中枢神经系统症状，并提供更广泛的 DM CNS中发生的所有转录组变化的全面视图。拟议的工作很重要，由于没有分子变化与DM CNS中的任何表型相关。这为未来的治疗努力旨在纠正中枢神经系统缺陷。