Transcriptional profiling in child mitochondrial disease

儿童线粒体疾病的转录谱

• 批准号: 6852108
• 负责人: WILLIAM James CRAIGEN
• 金额: $ 7.5万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2005
• 资助国家: 美国
• 起止时间: 2005-01-01 至 2006-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6852108
• 关键词: RNA; central nervous system disorders; clinical research; computer assisted sequence analysis; cytochrome c; cytochrome oxidase; fibroblasts; gene expression profiling; gene mutation; genetic transcription; human genetic material tag; lactic acidosis; microarray technology; mitochondrial disease /disorder; northern blottings; pediatrics; tissue /cell culture; western blottings

DESCRIPTION (provided by applicant): Mitochondrial encephalomyopathies are a group of genetically heterogeneous disorders for which molecular diagnosis remains remarkably difficult. One particular form of mitochondrial encepaholmyopathy termed Leigh syndrome (LS) is a pediatric neurodegenerative condition and is a phenotypic manifestation of a variety of disorders of energy metabolism, most commonly cytochrome c oxidase (COX) deficiency. Because of locus heterogeneity associated with the clinical phenotype, identifying the molecular basis for this disease and other mitochondrial disorders remains an enormous challenge. Currently, the state-of-the-art in evaluating patients with suspected mitochondrial disorders is to perform a muscle biopsy and carry out biochemical assays of the respiratory chain. However, it does not provide a specific gene diagnosis, which remains a research endeavor. The investigators would like to explore the feasibility of establishing specific diagnoses using transcriptional profiling of total RNA from cultured skin fibroblasts. It is hypothesized that there will be unique transcriptional profiles for a number of single gene mitochondrial disorders. To apply this to LS and other mitochondrial diseases, data sets of known gene defects need to be developed. Preliminary data from the investigator's laboratory using COX deficient fibroblasts with mutations in SURF1 demonstrates that cluster analysis identifies subsets of genes that are either up or down regulated in a consistent fashion. The investigators propose to validate these observations using additional SURF1 deficient fibroblasts by comparing the output using three different software packages: Rosetta, GeneSpring, and dChip. Triplicate comparison will be made and compared to a single wild-type control also analyzed in triplicate. The effect of culture conditions and passage number will be examined. Clinically suspected samples will be analyzed and gene predictions made based upon known patterns. In those disorders manifested in cultured cells, this approach offers the potential for a much less invasive means of determining the specific nuclear gene defect. These results will help in counseling, the development of potential therapies, and assessing long-term outcomes.

描述（由申请人提供）：线粒体脑肌病是一组遗传异质性疾病，分子诊断仍然非常困难。 线粒体脑肌病的一种特殊形式称为Leigh综合征（LS），是儿科神经退行性疾病，是多种能量代谢紊乱的表型表现，最常见的是细胞色素c氧化酶（考克斯）缺乏。 由于与临床表型相关的基因座异质性，确定这种疾病和其他线粒体疾病的分子基础仍然是一个巨大的挑战。 目前，评估疑似线粒体疾病患者的最新技术是进行肌肉活检并进行呼吸链的生化测定。 然而，它不提供特定的基因诊断，这仍然是一项研究奋进。 研究人员希望探索利用培养的皮肤成纤维细胞总RNA的转录谱建立特异性诊断的可行性。 据推测，将有一些单基因线粒体疾病的独特的转录谱。 为了将其应用于LS和其他线粒体疾病，需要开发已知基因缺陷的数据集。 研究者实验室使用SURF 1突变的考克斯缺陷成纤维细胞的初步数据表明，聚类分析确定了以一致方式上调或下调的基因子集。 研究人员建议使用额外的SURF1缺陷成纤维细胞通过比较三种不同软件包的输出来验证这些观察结果：Rosetta，GeneSpring和dChip。 将进行三次比较，并与同样一式三份分析的单个野生型对照进行比较。 将检查培养条件和传代次数的影响。 将对临床疑似样本进行分析，并根据已知模式进行基因预测。 在培养细胞中表现出的那些疾病中，这种方法提供了一种确定特定核基因缺陷的侵入性小得多的方法的潜力。 这些结果将有助于咨询，潜在疗法的开发和评估长期结果。