Identification of Genetic and Molecular Pathways in Congenital Rare Disorders Affecting the Brain and Muscle

影响大脑和肌肉的先天性罕见疾病的遗传和分子途径的鉴定

• 批准号: 10671469
• 负责人: M. CHIARA MANZINI
• 金额: $ 32.87万
• 依托单位: RUTGERS BIOMEDICAL AND HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-15 至 2024-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10671469
• 关键词: Affect; Biochemical; Biopsy; Brain; Childhood; Code; Cognitive deficits; Cohort Studies; Complex; Consumption; DNA Sequence Alteration; DNA sequencing; Development; Developmental Delay Disorders; Disease; Dystroglycan; Embryo; Extracellular Matrix; Eye; Family; Future; Genes; Genetic; Genetic Heterogeneity; Genetic Models; Genetic study; Genome; Genotype; Goals; Histologic; Human; Individual; Modeling; Molecular; Motor; Muscle; Muscle Cells; Muscle Development; Muscular Dystrophies; Mutate; Mutation; Neurons; Nucleic Acid Regulatory Sequences; Organ; Pathogenesis; Pathway interactions; Phenotype; Postdoctoral Fellow; Protein Glycosylation; RNA Splicing; Rare Diseases; Research; Research Personnel; Severities; Specificity; Testing; Therapeutic; Therapeutic Intervention; Therapy Evaluation; Time; Translations; Untranslated RNA; Variant; Work; Zebrafish; analysis pipeline; bioinformatics pipeline; clinical heterogeneity; cognitive disability; cohort; congenital muscular dystrophy; cost; data integration; deafness; disease-causing mutation; dystroglycanopathy; exome; gene discovery; genetic analysis; genetic architecture; genetic disorder diagnosis; genetic testing; genome sequencing; glycosylation; glycosyltransferase; in vivo Model; mortality; mouse model; next generation sequencing; novel; novel therapeutics; overexpression; patient population; targeted treatment; therapeutic evaluation; therapy development; transcriptome; transcriptome sequencing; treatment group; whole genome

ABSTRACT Congenital muscular dystrophies (CMDs) are a group of heterogeneous pediatric disorders leading to motor and developmental delay, and childhood mortality. CMDs have variable presentation often affecting multiple organs, such as the eyes and brain. While this clinical heterogeneity initially hindered genetic analyses, the ad- vent of next-generation sequencing and whole exome studies greatly increased gene identification reaching up to 30 genes, with eight genes identified in 2012-2013, three of which by the PI. Each new gene has provided a different piece of a complex puzzle, not only providing a genetic diagnosis for the affected individuals, but also informing us on how the mutated genes converge onto shared molecular pathways such as protein glycosyla- tion. Yet, our group and others have found that mutations in each gene are only present in a small portion of cases, and 30-40% of cases remain unexplained. In parallel, the large number of known genes has hindered therapy development as it remains unclear how cases with different genotypes and phenotypes can be grouped for treatment. The PI has focused the past decade on studying the genetics of CMD and developing zebrafish models to define how disease-causing mutations affect muscle and brain development. Through these studies, we have devel- oped our central hypotheses that most CMD genes regulate interactions with the extracellular matrix (ECM) through glycosylation, and that unexplained cases will either carry mutations in novel genes involved in cell- ECM interaction or noncoding variants in already known CMD genes. The proposed studies will test these hypotheses through two independent and complementary Specific Aims. Specific Aim 1 will leverage the multi- ple zebrafish models we have developed for known CMD genes to test whether increasing glycosylation will restore cell-ECM interactions in the muscle and brain in different genetic models of CMD. We will 1) define which biochemical deficits are shared by different genetic mutations in muscle cells and neurons, and 2) test whether changes in glycosylation can be beneficial in different forms of CMD. This workflow can then be ex- panded to future therapeutic interventions and novel disease genes can be rapidly taken from gene identifica- tion to therapy evaluation. Specific Aim 2 will close the gap in gene discovery in CMDs by testing the hypothe- sis that undiagnosed CMD cases are caused by a combination of rare mutations in novel genes and non- coding mutations in intronic/regulatory regions of known CMD genes. We have developed a next generation sequencing and bioinformatic pipeline that will integrate data from exome, genome, and transcriptome to iden- tify coding, splicing, and regulatory variants to fully unravel the genetics of CMD. These studies will directly impact the CMD fields by both discovering how different disease genes contribute to pathogenesis and by developing novel genetic tests for the global patient population.

抽象的 先天性肌营养不良症 (CMD) 是一组导致运动障碍的异质儿科疾病 发育迟缓和儿童死亡率。 CMD 具有可变的表现形式，通常会影响多个 器官，例如眼睛和大脑。虽然这种临床异质性最初阻碍了遗传分析，但 新一代测序和全外显子组研究的出现大大增加了基因鉴定 到 30 个基因，其中 8 个基因是在 2012-2013 年确定的，其中 3 个是 PI 鉴定的。每个新基因都提供了 复杂难题的不同部分，不仅为受影响的个体提供基因诊断，而且还 告诉我们突变基因如何汇聚到共享分子途径，例如蛋白质糖基化- 。然而，我们的小组和其他人发现每个基因的突变只存在于一小部分 且 30-40% 的病例仍无法解释。与此同时，大量的已知基因阻碍了 治疗的发展，因为目前尚不清楚如何对具有不同基因型和表型的病例进行分组 进行治疗。 在过去的十年里，PI 一直致力于研究 CMD 的遗传学并开发斑马鱼模型来定义 致病突变如何影响肌肉和大脑发育。通过这些研究，我们开发了 我们的中心假设是，大多数 CMD 基因调节与细胞外基质 (ECM) 的相互作用 通过糖基化，无法解释的病例要么携带与细胞相关的新基因突变 ECM 相互作用或已知 CMD 基因中的非编码变异。拟议的研究将测试这些 通过两个独立且互补的具体目标提出假设。具体目标 1 将利用多 我们为已知的 CMD 基因开发了多个斑马鱼模型，以测试增加糖基化是否会 在不同的 CMD 遗传模型中恢复肌肉和大脑中细胞与 ECM 的相互作用。我们将 1) 定义 肌肉细胞和神经元的不同基因突变共有哪些生化缺陷，2) 测试 糖基化的变化是否对不同形式的 CMD 有益。然后这个工作流程可以是ex- 适应未来的治疗干预措施，并且可以从基因鉴定中快速获取新的疾病基因 重刑治疗评估。具体目标 2 将通过测试假设来缩小 CMD 基因发现的差距 未确诊的 CMD 病例是由新基因的罕见突变和非基因突变共同引起的。 已知 CMD 基因的内含子/调节区域的编码突变。我们已经开发了下一代 测序和生物信息管道将整合外显子组、基因组和转录组的数据以识别 tify 编码、剪接和调控变异，以完全阐明 CMD 的遗传学。 这些研究将发现不同的疾病基因如何影响 CMD 领域，从而直接影响 CMD 领域。 发病机制并为全球患者群体开发新的基因测试。

项目成果

Novel mutation identification and copy number variant detection via exome sequencing in congenital muscular dystrophy.

• DOI: 10.1002/mgg3.1387
• 发表时间: 2020-11
• 期刊: Molecular genetics & genomic medicine
• 影响因子: 2
• 作者: Cauley ES;Pittman A;Mummidivarpu S;Karimiani EG;Martinez S;Moroni I;Boostani R;Podini D;Mora M;Jamshidi Y;Hoffman EP;Manzini MC
• 通讯作者: Manzini MC