Characterization of functional molecular domains of MeCP2

MeCP2 功能分子结构域的表征

基本信息

批准号：
10224679
负责人：
Bridget E Collins
金额：
$ 5.01万
依托单位：
VANDERBILT UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-08-01 至 2023-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10224679
关键词：
Address Adverse effects Affect Affinity Age Aggressive behavior Alleles Behavioral Binding Biological Assay Brain Brain region C-Terminal Binding Protein 2 C-terminal Cell model Cells Chromatin Communication Communities Complement Complementary DNA Complex Critical Thinking DNA DNA Binding Data Development Disease Disease model Dissection Environment Fellowship Functional disorder Gene Expression Genes Genetic Genetic Engineering Genetic Transcription Goals Hand High Prevalence In Vitro Individual Institutes Investigation Life Link MeCP2 Duplication Syndrome Methyl-CpG-Binding Protein 2 Missense Mutation Molecular Molecular Conformation Movement Mus Mutation Neurodevelopmental Disorder Overlearning Pathogenicity Patients Pediatric Neurology Phenotype Physiological Plasmids Point Mutation Proteins Reader Repressor Proteins Research Research Training Rett Syndrome Risk Severity of illness Speech System Technical Expertise Therapeutic Time Training Transgenic Organisms Work autistic clinical decision-making clinically relevant conditioned fear disability disease-causing mutation falls gain of function gain of function mutation gene therapy genetic corepressor girls in vivo innovation insight loss of function loss of function mutation molecular domain mouse model mutant novel novel therapeutic intervention pediatric department precision medicine preservation protein function restoration skills

项目摘要

PROJECT SUMMARY/ABSTRACT Rett syndrome, an X-linked neurodevelopmental disorder caused predominantly by mutations in the gene encoding chromatin modulator Methyl-CpG-binding protein 2 (MECP2), is a leading genetic cause of disability in girls worldwide. Affected individuals develop typically for a period of 6-18 months, at which time the disease causes developmental regression with loss of purposeful hand movements, loss of speech, and autistic features. Treatment options are currently limited to symptomatic management, making the development of novel therapeutic approaches critically important. 10% of disease-causing MECP2 mutations arise in its C- terminal domain. However, despite the high prevalence of mutations in this region, few studies have investigated their pathophysiological mechanism(s). Furthermore, proposed therapeutic strategies for RTT focus largely on remedying loss-of-function (LOF) mutations that occur in other domains of MeCP2 - the methyl-binding and NCoR-interaction domains. Previous work suggests that the C-terminus may alter local chromatin binding and conformation, but thorough functional studies of different types of C-terminal mutations are lacking. Preliminary data shows that one of the most common C-terminal mutations that eliminates the entire domain, R294X, yields a truncated protein product in mice that accumulates with age and binds chromatin more tightly than wild-type MeCP2, suggesting that the R294X mutation may not act through simple LOF. With the resulting hypothesis that some C-terminal mutations cause Rett syndrome via a non-LOF mechanism, this proposal aims to 1) determine the contributions of the MeCP2 C-terminus to protein function as a transcriptional modulator, 2) elucidate how the R294X mutation elicits transcriptional dysregulation in three different brain regions, and 3) establish the behavioral consequences of transgenic complementation of the R294X allele. The proposed studies will involve the development of an in vitro framework to probe C- terminal Mecp2 mutations in functional assays for chromatin binding and co-repressor interaction, as well as the use of dual molecular and behavioral approaches to dissect the pathogenic mechanism of the prevalent R294X mutation. This research will provide mechanistic insight into the complex neurodevelopmental disorder Rett syndrome, as well as assist clinical decision-making by identifying the viability of proposed therapies for individuals with C-terminal MECP2 mutations. The proposed research will be accomplished through a carefully crafted fellowship training plan that involves opportunities to develop the applicant's technical expertise, critical-thinking skills, and scientific communication skills. Further, the research environment provided in turns by the MSTP, Vanderbilt Brain Institute, and Department of Pediatric Neurology is ideal for the proposed research and training, harboring a reputation for scientific excellence, a collegial community, and a strong focus on trainee development.

项目概要/摘要 Rett 综合征，一种 X 连锁神经发育障碍，主要由基因突变引起编码染色质调节剂甲基 CpG 结合蛋白 2 (MECP2)，是导致残疾的主要遗传原因在全世界的女孩中。受影响的个体通常会持续 6-18 个月的时间，在此期间疾病会导致发育倒退，丧失有目的的手部动作、语言能力丧失和自闭症特征。目前的治疗选择仅限于对症治疗，使得新颖的治疗方法至关重要。 10% 的致病 MECP2 突变出现在其 C- 终端域。然而，尽管该区域的突变发生率很高，但很少有研究表明研究其病理生理机制。此外，提出的 RTT 治疗策略主要关注于补救 MeCP2 其他域中发生的功能丧失 (LOF) 突变 - 甲基结合域和 NCoR 相互作用域。先前的工作表明 C 末端可能会改变局部染色质结合和构象，但对不同类型的 C 端突变进行彻底的功能研究缺乏。初步数据显示，最常见的 C 端突变之一消除了整个结构域 R294X 在小鼠体内产生截短的蛋白质产物，该产物随着年龄的增长而积累并结合染色质比野生型 MeCP2 更紧密，表明 R294X 突变可能无法通过简单的作用发挥作用洛夫。由此产生的假设是，一些 C 端突变通过非 LOF 导致 Rett 综合征机制，该提案旨在 1) 确定 MeCP2 C 末端对蛋白质功能的贡献作为转录调节剂，2) 阐明 R294X 突变如何引起转录失调三个不同的大脑区域，3）确定转基因互补的行为后果 R294X 等位基因。拟议的研究将涉及开发一个体外框架来探测 C- 染色质结合和共阻遏物相互作用的功能测定中的末端 Mecp2 突变，以及使用双分子和行为方法来剖析流行病的致病机制 R294X 突变。这项研究将为复杂的神经发育障碍提供机制上的见解 Rett 综合征，以及通过确定拟议疗法的可行性来协助临床决策 C 端 MECP2 突变的个体。拟议的研究将通过仔细的研究来完成精心设计的奖学金培训计划，其中包括发展申请人技术专长的机会，批判性思维能力和科学沟通能力。进一步，轮流提供研究环境由 MSTP、范德比尔特脑研究所和小儿神经病学系联合开发的项目非常适合拟议的研究和培训，享有科学卓越、大学社区和强大的声誉注重学员发展。