Identifying Pathogenic Non-Coding Mutations in Rare Mendelian Disease

鉴定罕见孟德尔病的致病性非编码突变

• 批准号: 9806572
• 负责人: GREGORY E CRAWFORD
• 金额: $ 19.65万
• 依托单位: DUKE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2019
• 资助国家: 美国
• 起止时间: 2019-08-07 至 2021-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9806572
• 关键词: Affect; Alleles; Amino Acid Sequence; Biochemical; Biological Assay; Biological Markers; Biomedical Engineering; Biopsy Specimen; Blood specimen; Cell Line; Cell model; Cessation of life; Childhood; Clinical; Clustered Regularly Interspaced Short Palindromic Repeats; Code; Custom; DNA; DNA Sequence; Data; Deficiency Diseases; Development; Diagnosis; Diagnostic tests; Dideoxy Chain Termination DNA Sequencing; Disease; Early Diagnosis; Early Intervention; Early treatment; Enhancers; Exons; Family; Foundations; Frameshift Mutation; Gene Expression; Genes; Genetic; Genetic Counseling; Genetic screening method; Genomics; Glycogen Storage Disease; Goals; Health; Healthcare Systems; Impairment; Institutional Review Boards; Insurance; Introns; Lead; Life; Long-Chain-Acyl-CoA Dehydrogenase; Mendelian disorder; Methods; Mutation; Outcome; Pathogenicity; Patients; Protein Isoforms; Proteins; RNA Splicing; Rare Diseases; Reagent; Regulation; Regulator Genes; Regulatory Element; Research Personnel; Sampling; Site; Splice-Site Mutation; Techniques; Test Result; Testing; Therapeutic Intervention; Untranslated RNA; VLCAD deficiency; Variant; World Health; acyl-CoA dehydrogenase; clinical diagnostics; design; disease-causing mutation; early onset; effective therapy; epigenetic regulation; experimental study; fatty acid oxidation; genetic variant; genome editing; improved; innovation; novel; prevent; protein function; statistics; transcriptome sequencing

ABSTRACT Determining new causes for rare and common disease would have major and immediate benefits for patients and their families by improved genetic testing, genetic counseling, insurance reimbursement, and ultimately more effective treatment options. Our long term goal is to disruptively improve and expand genetic testing for rare and common disease. Current diagnostic tests only consider pathogenic variants in protein-coding genes. However, we now have evidence that a substantial fraction of rare disease is due to unknown non-coding genetic variants that influence the regulation of those genes. The goal of this proposal is to identify and quantify the effect of pathogenic non-coding genetic variants on the function and expression of genes that cause rare disease. This initial step will enable treatment early in life when it is still possible to stop the most severe consequences of disease, including death. We will focus on severe early-onset pediatric disorders, including glycogen storage diseases (GSD I, II, III, IV, and IX), and the fatty acid oxidation disorders, very long-chain acyl-CoA dehydrogenase deficiency (VLCAD), and multiple acyl-CoA dehydrogenase deficiency (MADD). To date, genetic tests for these and other diseases are limited to protein-coding mutations. However, our clinical team has collected numerous cases that have a single pathogenic coding variant on only one of the two alleles that must be both affected in these recessive disorders. We also have biochemical and biomarker evidence that supports the diagnosis. Those cases are an ideal opportunity to identify additional disease-causing variants. Our hypothesis is that the genetic causes of recessive disorders include novel genetic variants that can alter either protein sequence (Aim 1), splicing (Aim 2), or gene expression (Aim 3) of disease genes. We have assembled a team of Pediatric clinicians who are experts in GSDs, VLCAD, and MADD, as well as researchers who are experts in genetics, genomics, epigenetic regulation, biomedical engineering, and statistics. This team has obtained patient samples and received Duke IRB approval to begin immediately. We expect this study will identify and validate novel genetic variants that influence disease. While we propose to study a relatively small subset of rare disorders, these strategies will be immediately generalizable to any patient sample with any recessive disorder that has inconclusive genetic testing results. That outcome will provide comprehensive genetic testing, better understanding of disease mechanisms, and ultimately better treatment options.

摘要 确定罕见和常见疾病的新病因将有重大而直接的好处 通过改进的基因检测、遗传咨询、保险为患者及其家人提供 报销，并最终实现更有效的治疗选择。我们的长期目标是 颠覆性地改进和扩大罕见病和常见病基因检测。当前诊断 测试只考虑蛋白质编码基因中的致病变异。然而，我们现在有证据表明 很大一部分罕见疾病是由未知的非编码遗传变异引起的 影响这些基因的调控。这项提议的目标是确定和量化 致病性非编码基因变异对血管紧张素转换酶功能和表达的影响 导致罕见疾病的基因。这一最初的步骤将使在生命早期接受治疗成为可能。 有可能阻止疾病造成的最严重后果，包括死亡。我们将重点关注 严重的早发性儿科疾病，包括糖原储存疾病(GSD I、II、III、IV和 和脂肪酸氧化紊乱，超长链酰辅酶A脱氢酶缺乏症 (VLCAD)和多发性酰辅酶A脱氢酶缺乏症(MADD)。到目前为止，基因测试 这些疾病和其他疾病仅限于蛋白质编码突变。然而，我们的临床团队已经 收集了大量的病例，这些病例只在两种病毒中的一种上有一个致病编码变体 在这些隐性疾病中必须同时受到影响的等位基因。我们还有生化和 支持诊断的生物标志物证据。这些案件是确定 更多的致病变异。我们的假说是隐性遗传的原因 疾病包括可以改变蛋白质序列(目标1)、剪接等的新型遗传变异 (目标2)，或疾病基因的基因表达(目标3)。我们已经组建了一支儿科团队 临床医生是GSD、VLCAD和MADD方面的专家，以及研究人员是专家 遗传学、基因组学、表观遗传调控、生物医学工程和统计学。这支队伍 已获得患者样本，并获得杜克大学IRB批准立即开始。我们预计 这项研究将识别和验证影响疾病的新基因变异。当我们提议 为了研究相对较小的罕见疾病子集，这些策略将立即 可推广到任何具有不确定遗传的隐性疾病的患者样本 检测结果。这一结果将提供全面的基因测试，更好地了解 疾病机制，以及最终更好的治疗选择。