RNA Processing-Mediated Mechanisms of CNS Dysfunction in Myotonic Dystrophy

强直性肌营养不良中 CNS 功能障碍的 RNA 加工介导机制

基本信息

批准号：
10652533
负责人：
GARY J BASSELL
金额：
$ 55.73万
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 2025-06-30
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10652533
关键词：
3&apos Untranslated Regions Adult Affect Alternative Splicing Animal Model Antisense Oligonucleotide Therapy Antisense Oligonucleotides Autopsy Behavioral Biological Markers Biology Brain Brain imaging CUG repeat Central Nervous System Cerebrospinal Fluid Childhood Clinical Trials Cognitive Complex Mixtures Defect Diagnosis Disease Disease Marker Disease Progression Dissection Dominant Genetic Conditions Dopamine D2 Receptor Down Syndrome Drowsiness Equilibrium Event Excitatory Synapse Exhibits Exons FDA approved Fatigue Fragile X Syndrome Functional disorder Future Gene Expression Profiling Genetic Diseases Glutamates Goals Heart Human Huntington Disease Image Impairment Inherited Inhibitory Synapse Knockout Mice Link Measures Mediating Methods Modality Modeling Molecular Mus Muscle Muscle Weakness Muscle function Myotonia Myotonic Dystrophy NMDA receptor A1 Neurons Neurotransmitters Patients Persons Pharmaceutical Preparations Phenotype Physiological Play Poly A Polyadenylation Post-Transcriptional Regulation Prevalence Protein Isoforms Protein Kinase Proteins Public Health RNA RNA Processing RNA Splicing RNA-Binding Proteins Raclopride Research Role Skeletal Muscle Sleep disturbances Spliced Genes Symptoms Synapses Systemic disease Therapeutic Tissues Transcript Transgenic Mice Vertebrates Work autism spectrum disorder autosome behavioral phenotyping biomarker development biomarker driven brain dysfunction brain tissue cell type clinical phenotype efficacy evaluation experience gamma-Aminobutyric Acid gene therapy genome-wide human data improved insight molecular phenotype mouse model nervous system disorder neurophysiology novel therapeutic intervention postsynaptic presynaptic receptor restoration synaptic function targeted agent targeted treatment transcriptome transcriptome sequencing wasting

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.

强直性肌营养不良（dys营养不良，DM）是一种常染色体显性遗传病，诊断为强直性肌营养不良。全世界有 1:8000 人患病，影响身体的多个组织，包括骨骼肌，心脏，以及与此提议相关的中枢神经系统（CNS）。 DM1是由CTG扩张引起的在肌强直营养不良蛋白激酶 (DMPK) 的 3' UTR 中重复。老鼠身上有大量证据 DM1 模型和人类 DM1 死后组织支持 RNA 介导的疾病机制，其中有毒的核内 CUG RNA 灶隔离器 Muscleblind (MBNL) RNA 结合蛋白，通常发挥作用在调节转录后基因调控的各个方面发挥着至关重要的作用。我们的一个重大差距我们的理解是，我们不知道哪些 RNA 加工缺陷是 DM1 特定损伤的基础大脑功能。最近的工作和我们的新发现表明，编码 RNA 的错误剪接突触蛋白是导致 DM1 中 CNS 功能障碍的原因。我们的中心假设是 CNS 表型直接归因于 MBNL 介导的 RNA 加工的丧失以及 MBNL 活性的恢复和/或剪接可以恢复大脑功能。我们的目标是全面了解 RNA 加工介导的 DM1 中枢神经系统功能障碍的机制，并利用它来开发和严格评估新的治疗方法策略。该提案的总体目标是使用候选方法和全基因组方法，应用于 MBNL KO 小鼠和基于新 AAV9 的神经元小鼠模型，与 RNAseq 分析相比人类死后大脑，评估特定剪接事件驱动症状的作用，并全面识别错误剪接和 RNA 加工的变化。目标 1 将描述如何 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 将开发一种新的基于 AAV9 的小鼠模型来阐明 RNA 组通过转录谱和重叠处理神经元中导致 DM1 表型的事件人类 DM 大脑与 DM 小鼠模型大脑。目标 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 鼠标型号。这些研究将为扰动如何影响特定的机制提供新的机制见解。 RNA 加工事件可导致强直性肌营养不良的中枢神经系统症状，并提供更广泛的研究 DM CNS 中发生的所有转录组变化的全面视图。拟议的工作意义重大，因为没有分子变化与 DM CNS 中的任何表型相关。这提供了框架未来的治疗努力旨在纠正中枢神经系统缺陷。