RNA Dominance in Human Disease

RNA 在人类疾病中的主导地位

• 批准号: 7600482
• 负责人: MAURICE SCOTT SWANSON
• 金额: $ 29.74万
• 依托单位: UNIVERSITY OF FLORIDA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-04-01 至 2010-08-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7600482
• 关键词: 3' Untranslated Regions; Adult; Affect; Alleles; Alternative Splicing; Antibodies; Appearance; Behavioral; Benign congenital hypotonia; Binding; Binding Proteins; Binding Sites; Biochemical; Biological Assay; Brain; Cardiac; Cataract; Cell Nucleus; Characteristics; Complex; Defect; Development; Disease; Double-Stranded RNA; Embryo; Embryonic Development; Event; Exons; Family; Fluorescent in Situ Hybridization; Functional RNA; Genes; Genomics; Goals; Grant; Human; Immunofluorescence Immunologic; Immunoprecipitation; Infant; Inherited; Introns; Knock-out; Knockout Mice; Length; Limb structure; Link; Mental Retardation; Microsatellite Repeats; Modeling; Molecular; Monoclonal Antibodies; Mus; Muscle; Muscle Development; Muscle hypotonia; Muscle relaxation phase; Muscular Dystrophies; Mutation; Myoblasts; Myotonia; Myotonic Dystrophy; Neonatal; Nervous system structure; Neuromuscular Diseases; Nuclear RNA; Pathogenesis; Pattern; Phenotype; Plasmids; Procedures; Process; Protein Family; Protein Isoforms; Proteins; RNA; RNA Processing; RNA Splicing; RNA-Binding Proteins; Recruitment Activity; Regulation; Reporting; Research; Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; Site; Staging; Structure; Testing; Tissues; Transcript; Transfection; Transgenes; Transgenic Mice; Transgenic Model; base; crosslink; early onset; gain of function; human disease; loss of function; mRNA Precursor; mouse model; mutant; myogenesis; neurogenesis; novel; novel therapeutics; overexpression; postnatal; prevent; programs; wasting

DESCRIPTION (provided by applicant): Although myotonic dystrophy (DM) is the most prevalent muscular dystrophy in adults, a more severe form of this neuromuscular disease is congenital DM (COM). While the two characterized types of adult-onset DM, DM1 and DM2, are caused by unstable microsatellite (CTG)n and (CCTG)n expansions in the DMPK and ZNF9 genes, respectively, CDM is only associated with >1,000 CTG repeats in the DMPK gene. The longterm goals of the proposed research are to elucidate the molecular events which result in CDM and use this information to develop novel therapeutic strategies for this disease. According to the RNA dominance model, DM is an RNA gain-of-function disease in which mutant DMPK and ZNF9 RNAs are retained in the nucleus where they inhibit the functions of the muscleblind-like (MBNL) proteins. MBNL proteins are alternative splicing factors and current evidence suggests that DM disease is caused by the retention of neonatal protein isoforms in the adult. The objective of this proposal is to use both knockout and transgenic mouse models to extend the RNA dominance model to CDM using four experimental aims. First, the hypothesis that specific Mbnl proteins are sequestered during embryogenesis by (CUG)n expansions will be tested using a new transgenic mouse model for CDM. Second, CDM is characterized by neonatal hypotonia and mental retardation so potential roles for Mbnl proteins in the regulation of pre-mRNA processing during embryonic muscle and brain development will be investigated. Third, CDM is only associated with DMPK CTG repeat expansions and previous studies have reported changes in the expression of DMPK, as well as the linked genes DMWD and SIX5, in CDM. Therefore, several mouse knockout lines will be generated to test the hypothesis that CDM results from coordinate loss of MBNL and one, or several, of these DMPK associated genes. Fourth, overexpression of DMPK RNA inhibits myogenic differentiation so the hypothesis that CDM results from co-sequestration of MBNL and DMPK RNA-binding proteins will be evaluated.

描述（由申请人提供）：虽然肌强直性营养不良（DM）是成人中最常见的肌肉营养不良，但这种神经肌肉疾病的更严重形式是先天性DM （COM）。虽然两种典型的成人发病型糖尿病DM1和DM2分别是由DMPK和ZNF9基因中不稳定微卫星（CTG）n和（CCTG）n扩增引起的，但CDM仅与DMPK基因中CTG重复数的1000倍相关。提出的研究的长期目标是阐明导致CDM的分子事件，并利用这些信息开发新的治疗策略。根据RNA优势模型，糖尿病是一种RNA功能获得性疾病，其中突变的DMPK和ZNF9 RNA保留在细胞核中，在那里它们抑制了肌盲样（MBNL）蛋白的功能。MBNL蛋白是一种可选择的剪接因子，目前的证据表明，糖尿病是由新生儿蛋白同种异构体在成人体内的保留引起的。本提案的目的是利用敲除和转基因小鼠模型，通过四个实验目的将RNA优势模型扩展到CDM。首先，我们将使用一种新的CDM转基因小鼠模型来验证特异性Mbnl蛋白在（CUG）n扩增过程中被隔离的假设。其次，CDM以新生儿张力低下和智力低下为特征，因此Mbnl蛋白在胚胎肌肉和大脑发育过程中调控pre-mRNA加工的潜在作用将被研究。第三，CDM仅与DMPK CTG重复扩增相关，先前的研究报道了CDM中DMPK以及相关基因DMWD和SIX5的表达变化。因此，将产生几个小鼠敲除系来验证CDM是由MBNL和这些DMPK相关基因中的一个或几个的坐标丢失引起的假设。第四，DMPK RNA的过表达抑制了肌源性分化，因此CDM是由MBNL和DMPK RNA结合蛋白共同隔离引起的假说将被评估。