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）：虽然强直性肌营养不良（DM）是成人中最常见的肌营养不良，但这种神经肌肉疾病的更严重形式是先天性DM（COM）。虽然两种特征性类型的成人发病DM，DM 1和DM 2，分别由DMPK和ZNF 9基因中不稳定的微卫星（CTG）n和（CCTG）n扩增引起，但CDM仅与DMPK基因中> 1，000个CTG重复相关。该研究的长期目标是阐明导致CDM的分子事件，并利用这些信息来开发这种疾病的新治疗策略。根据RNA显性模型，DM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，在那里它们抑制肌盲样（MBNL）蛋白的功能。MBNL蛋白是选择性剪接因子，目前的证据表明，DM疾病是由新生儿蛋白异构体在成人中的保留引起的。本提案的目的是使用基因敲除和转基因小鼠模型，使用四个实验目标将RNA优势模型扩展到CDM。首先，将使用用于CDM的新的转基因小鼠模型来测试特定Mbnl蛋白在胚胎发生期间通过（CUG）n扩增被隔离的假设。第二，CDM的特征在于新生儿张力减退和智力迟钝，因此将研究Mbnl蛋白在胚胎肌肉和脑发育期间调节前mRNA加工的潜在作用。第三，CDM仅与DMPK CTG重复扩增相关，并且先前的研究已经报道了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以测试CDM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的过表达抑制肌原性分化，因此将评估CDM由MBNL和DMPK RNA结合蛋白的共螯合引起的假设。