DE PEDIATRIC COBRE: MOLECULAR MECHANISMS IN PELIZAEUS MERZBACHER DISEASE

DE PEDIATRIC COBRE：Pelizaeus Merzbacher 病的分子机制

• 批准号: 7720951
• 负责人: Grace M. Hobson
• 金额: $ 14.35万
• 依托单位: ALFRED I. DU PONT HOSP FOR CHILDREN
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-08-01 至 2009-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7720951
• 关键词: 5' Splice Site; Accounting; Affect; Alternative Splicing; Biological Assay; Childhood; Computer Retrieval of Information on Scientific Projects Database; Computer Simulation; Coupled; Defect; Diagnostic Procedure; Disease; ES Cell Line; Engineering; Enhancers; Exons; Family; Funding; Gene Expression; Gene Transfer; Genes; Genomics; Grant; Hereditary Disease; Institution; Introns; Link; Location; Molecular; Molecular Cytogenetics; Mus; Mutation; Myelin Proteins; Oligodendroglia; Pathogenesis; Pelizaeus-Merzbacher Disease; Play; Point Mutation; Protein Binding; Proteins; Proteolipids; RNA Splicing; Relative (related person); Research; Research Personnel; Resources; Role; Source; Structure; Testing; Trans-Activators; Transfer RNA; United States National Institutes of Health; cis acting element; disease phenotype; dosage; improved; leukodystrophy; novel; size; 5' Splice Site; Accounting; Affect; Alternative Splicing; Biological Assay; Childhood; Computer Retrieval of Information on Scientific Projects Database; Computer Simulation; Coupled; Defect; Diagnostic Procedure; Disease; ES Cell Line; Engineering; Enhancers; Exons; Family; Funding; Gene Expression; Gene Transfer; Genes; Genomics; Grant; Hereditary Disease; Institution; Introns; Link; Location; Molecular; Molecular Cytogenetics; Mus; Mutation; Myelin Proteins; Oligodendroglia; Pathogenesis; Pelizaeus-Merzbacher Disease; Play; Point Mutation; Protein Binding; Proteins; Proteolipids; RNA Splicing; Relative (related person); Research; Research Personnel; Resources; Role; Source; Structure; Testing; Trans-Activators; Transfer RNA; United States National Institutes of Health; cis acting element; disease phenotype; dosage; improved; leukodystrophy; novel; size

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Pelizaeus-Merzbacher disease (PMD), an X-linked leukodystrophy, is caused by defects of the proteolipid protein 1 gene (PLP1) that encodes the major CNS myelin protein. Approximately 60% of families have a genomic duplication that includes the PLP1 gene, and 15-20% of families have PLP1 point mutations. Studies in mice have shown that an increased dosage of PLP1 can account for disease pathogenesis. Further, studies have shown that mutations in noncoding regions can alter PLP1 expression levels or the ratio of the alternatively spliced forms PLP1 and DM20. Our long-term objective is to understand the molecular mechanisms involved in generating the PMD phenotype so rational treatments and improved diagnostic techniques can be developed. In the first aim, results on location, size, structure and sequence at the junctions of PMD duplications support the hypothesis that most are caused by a novel coupled homologous and nonhomologous mechanism. We will continue a combined cytogenetic, molecular, and in silico approach to refine our understanding of the mechanism. In the second aim, the hypothesis that the duplication structure affects expression of PLP1 is being tested by engineering different duplications into ES cell lines and analyzing the structural effects on gene expression before and after differentiation into oligodendrocytes. In the third aim, the hypothesis that some PMD mutations dysregulate splicing of PLP1 is being tested by using gene transfer and RNA-protein binding assays to investigate cis-acting elements and trans-acting factors involved in PLP1/DM20 alternative splicing. We have determined that exon and intron splicing enhancers and the relative strength of the PLP1 and DM20 donor splice sites play an important role in PLP1 alternative splicing. The results of these studies have greatest impact potential through their generalizability to other genomic and splicing diseases, which have only recently been recognized as important types of genetic disease.

该副本是利用众多研究子项目之一 由NIH/NCRR资助的中心赠款提供的资源。子弹和 调查员（PI）可能已经从其他NIH来源获得了主要资金， 因此可以在其他清晰的条目中代表。列出的机构是 对于中心，这不一定是调查员的机构。 Pelizaeus-Merzbacher疾病（PMD）是一种X连锁的白细胞营养不良，是由编码主要CNS髓磷脂蛋白的蛋白质蛋白1基因（PLP1）的缺陷引起的。大约60％的家庭具有包括PLP1基因在内的基因组重复，而15-20％的家庭具有PLP1点突变。在小鼠中的研究表明，PLP1的剂量增加可以解释疾病的发病机理。此外，研究表明，非编码区域的突变可以改变PLP1表达水平或替代剪接形式PLP1和DM20的比率。我们的长期目标是了解产生PMD表型所涉及的分子机制，以便可以开发出合理治疗和改进的诊断技术。在第一个目标中，PMD重复连接处的位置，大小，结构和序列的结果支持以下假设：大多数是由新型的同源和非同源机制引起的。我们将继续一种结合细胞遗传学，分子和硅方法，以完善我们对机制的理解。在第二个目标中，通过将不同的重复重复化为ES细胞系，并在分化为少突胶质细胞之前和之后分析了对基因表达的结构效应，从而测试了重复结构影响PLP1表达的假设。在第三个目标中，通过使用基因转移和RNA蛋白结合测定法研究了PLP1的某些PMD突变失调的剪接的假设，以研究参与PLP1/DM20替代旋转涉及的顺式作用元件和涉及的反式作用因子。我们已经确定，外显子和内含子剪接增强子以及PLP1和DM20供体剪接位点的相对强度在PLP1替代剪接中起重要作用。这些研究的结果通过对其他基因组和剪接疾病的推广性具有最大的影响潜力，这些疾病直到最近才被公认为是重要类型的遗传疾病。