Personalized Functional Genomics for Mitochondrial Encephalopathy Gene Discovery

线粒体脑病基因发现的个性化功能基因组学

• 批准号: 10582623
• 负责人: Penelope E Bonnen
• 金额: $ 65.92万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2014
• 资助国家: 美国
• 起止时间: 2014-08-20 至 2025-12-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10582623
• 关键词: Affect; Area; Ataxia; Automobile Driving; Bioinformatics; Biological; Biological Assay; Biological Markers; Cell model; Cells; Child; Childhood; Chronic; Chronic progressive external ophthalmoplegia; Clinical; Communities; Complement; Complementary DNA; Complex; Consensus; Coupled; DNA; DNA sequencing; Data; Data Analyses; Data Security; Data Set; Databases; Defect; Development; Diagnosis; Diagnostic; Disease; Disease model; Electron Transport; Encephalopathies; Etiology; Evaluation; Exhibits; Experimental Models; Functional disorder; Genes; Genetic; Genome; Genomic approach; Guidelines; Huntington Disease; Incidence; Individual; Inherited; International; Knock-in; Laboratories; Lentivirus Vector; Measures; Membrane Potentials; Mitochondria; Mitochondrial DNA; Mitochondrial Diseases; Molecular; Molecular Diagnosis; Multiomic Data; Muscle hypotonia; Mutagenesis; Mutation; Nerve Degeneration; Neurodegenerative Disorders; Neurologic; Nuclear; Ontology; Parkinson Disease; Pathogenesis; Pathogenicity; Pathology; Pathway interactions; Patients; Phenotype; Public Domains; RNA; Reactive Oxygen Species; Resources; Seizures; Series; Software Validation; Study Subject; System; Technology; Testing; Therapeutic; Validation; Variant; Visualization; Work; analysis pipeline; analytical method; analytical tool; biobank; body system; clinical phenotype; computational reasoning; data integration; data mining; disease-causing mutation; effective therapy; empowerment; exome; experience; functional genomics; gene discovery; genetic variant; genome sequencing; genome-wide; global health; heterogenous data; human disease; improved; innovation; insight; knock-down; knowledge base; metabolome; metabolomics; mitochondrial dysfunction; mitochondrial membrane; molecular diagnostics; multiple omics; nervous system disorder; neuromuscular; novel; transcriptome sequencing; transcriptomics; variant of unknown significance; virulence gene; whole genome

Project Summary Mitochondrial disease is a commonly occurring inherited condition, incidence 1/5000, which can affect every organ system and thus exhibits a broad range of clinical phenotypes. The most common are neurological and neuromuscular dysfunction that manifest as neurodegeneration, seizures, ataxia, chronic progressive external opthalmoplegia (CPEO), and hypotonia. Childhood-onset mitochondrial disease most often results from mutations in the nuclear genome; however, the majority of cases remain without a molecular diagnosis and no effective treatments thus underscoring the critical need to identify the genetic aberrations driving these disorders. We propose a personalized functional genomics approach combining genome-wide sequencing, transcriptomics, metabolomics and mitochondrial functional profiling in cells to identify validated novel mitochondrial disease genes and variants. We will leverage a multi-omic strategy for identifying the pathogenic genes and elucidating pathomechanisms: 1. Genome-wide sequencing of patients coupled with transcriptomics and metabolomics 2. Cell-based functional studies of genes and pathways identified in patients. Through our international network of collaborators we have collected patients with clinically confirmed primary mitochondrial encephalopathy who do not have a molecular diagnosis. For patients who have already had WES/WGS but no molecular diagnosis we will re-interpret these data and leverage our ability to interpret beyond ABMGG guidelines for diagnosis. Additionally, we have a parallel effort to identify disease genes through datamining the clinical exome database at Baylor Genetics diagnostic laboratory wherein genes that are known to be essential for mitochondrial function but are not yet demonstrated as disease causing are analyzed for mutations in patients. Gene causality will be determined through a series of cell-based disease modeling experiments of mitochondrial functional profiling that include strategies of gene knock down, high-throughput mutagenesis knock-in, and cDNA complementation studies. We will utilize this technology to test the functionality of variants of uncertain significance identified in our sequencing efforts as well as those obtained through collaborators, diagnostic laboratories, and the public domain. This work will generate an unprecedented resource of systematic profiling of cellular mitochondrial function and functionally- confirmed pathogenic molecular defects. The elucidation of these pathogenic genes and variants will immediately improve the molecular diagnostic potential for children with suspected mitochondrial disease. Moreover, by identifying the pathogenic genes for primary mitochondrial encephalopathy we will empower the scientific community focused on neurological and neurodegenerative disorders, which have a more complex etiology, by delivering genes and pathways for further study of the pathogenetic mechanisms of these global health problems.

项目摘要 线粒体疾病是一种常见的遗传性疾病，发病率为1/5000，可以影响每一个人。 器官系统，因此表现出广泛的临床表型。最常见的是神经系统疾病， 神经肌肉功能障碍，表现为神经变性、癫痫发作、共济失调、慢性进行性 眼外肌麻痹（CPEO）和张力减退。儿童期发病的线粒体疾病最常导致 核基因组突变;然而，大多数病例仍然没有分子诊断 没有有效的治疗方法，因此强调迫切需要确定驱动这些疾病的遗传畸变。 紊乱我们提出了一种个性化的功能基因组学方法， 转录组学、代谢组学和线粒体功能分析，以鉴定经验证的新的 线粒体疾病基因和变体。我们将利用多组学策略来识别 致病基因和阐明病理机制：1.患者的全基因组测序加上 转录组学和代谢组学2.基于细胞的基因和途径的功能研究， 患者通过我们的国际合作者网络，我们收集了临床 确诊的原发性线粒体脑病，但没有分子诊断。的患者 已经进行了WES/WGS，但没有分子诊断，我们将重新解释这些数据，并利用我们的 能够解释ABMGG诊断指南以外的内容。此外，我们还同时努力确定 通过对贝勒遗传学诊断实验室的临床外显子组数据库进行数据挖掘， 其中已知对于线粒体功能是必需的但尚未被证实为 分析患者中的突变。基因因果关系将通过一系列的 线粒体功能谱的基于细胞的疾病建模实验，包括基因调控策略 敲低、高通量诱变敲入和cDNA互补研究。我们会利用这个 技术来测试在我们的测序工作中鉴定的不确定意义的变体的功能， 以及通过合作者、诊断实验室和公共领域获得的那些。这项工作将 产生了前所未有的细胞线粒体功能和功能的系统分析资源- 确认了致病分子缺陷这些致病基因和变异的阐明将 立即提高疑似线粒体疾病儿童的分子诊断潜力。 此外，通过鉴定原发性线粒体脑病的致病基因，我们将使 科学界专注于神经和神经退行性疾病，这些疾病具有更复杂的 病原学，通过提供基因和途径，进一步研究这些全球性的发病机制， 健康问题