Large-scale functional phenotyping of ion channel arrhythmia genomic variants

离子通道心律失常基因组变异的大规模功能表型分析

• 批准号: 8757581
• 负责人: THOMAS V MCDONALD
• 金额: $ 15.14万
• 依托单位: ALBERT EINSTEIN 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: 2014-08-07 至 2016-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8757581
• 关键词: Affect; Arrhythmia; Awareness; Behavior; Behavior Therapy; Biochemical; Biological Assay; Biology; Cardiac; Cardiovascular system; Cell physiology; Clinical; Clinical Management; Complementary DNA; Computer Simulation; DNA; DNA Sequence; Databases; Defect; Defibrillators; Diagnosis; Disease; Disease susceptibility; Electrophysiology (science); Exhibits; Explosion; Family member; Generations; Genes; Genetic; Genetic Databases; Genetic Polymorphism; Genetic screening method; Genome; Genomics; Goals; Healthcare Systems; Heart Diseases; Hereditary Disease; Human; Human Cell Line; Implant; Inherited; Internet; Ion Channel; Ion Channel Protein Gene; Lead; Life; Life Style; Literature; Mission; Modification; Mutation; National Heart, Lung, and Blood Institute; Online Systems; Pacemakers; Patient Self-Report; Patients; Pharmaceutical Preparations; Phenotype; Physicians; Population; Procedures; Proteins; Reporting; Research Personnel; Resources; Risk; Side; Site; Source; Surface; Susceptibility Gene; Symptoms; System; Technology; Testing; Time; United States National Institutes of Health; Variant; Work; cDNA Expression; clinical phenotype; cohort; disorder risk; exome; functional genomics; genetic variant; heart rhythm; implantation; improved; loss of function; mutant; next generation sequencing; public health relevance; response; sudden cardiac death; trafficking

DESCRIPTION (provided by applicant): This proposal is in response to the RFA-HL-13-027: "Functional Assays to Screen Genomic Hits" (R21/R33). In the past 15 years over 40 genetic loci have been identified as associated with increased risk for cardiac arrhythmia and/or sudden cardiac death with Mendelian and multigenic inheritance. With the advance of rapid clinical genetic testing and Next-Generation sequencing, the number of mutations/variants has grown to greater than 1000 for the top loci alone (LQT1-3). A large majority of these variants have not been functionally phenotyped however. This poses a considerable dilemma for the clinical geneticist and managing cardiologist-are uncharacterized variants/mutations to be treated as true risks for arrhythmia? This decision is critical because therapy includes life-long medication, behavioral modification and often implantation of cardiac defibrillators. Moreover, these management decisions extend to all other family members harboring the variants. Furthermore, these clinical scenarios will soon become increasingly common as rapid, accessible and affordable whole exome/genome takes its place in the clinical arena. Till now, functional phenotyping of new variants has been performed in piecemeal fashion, with reports of one or a handful of variants at a time. This usually takes place as a side-project for investigators who are pursuing other aspects of cardiac biology. We propose to functionally phenotype all of the disease-associated genetic variants and most of the rare polymorphisms that have appeared in genetic databases for the loci most commonly reported for hereditary arrhythmias. Variants will be created in cDNAs for expression in human cell lines. Functional expression will be assessed with a high-throughput electrophysiology system. Subsequently, those variants exhibiting a loss-of-function phenotype will be further assayed for surface trafficking defects by biochemical analysis. We will create a publically available database with both ongoing and completed results that will allow other investigators to provide input. Creation of this public resource will provide clinical geneticist and managing physicians a valuable resource to fully analyze the results from genetic testing of their patients. Successful competition of this project will provide a framework and resource for investigators in the field and have immediate impact on clinical management of an increasing number of patients. Accordingly we believe that this proposal is in the spirit of both the RFA-HL- 13-027 and the mission of the NIH/NHLBI.

描述（由申请人提供）：本提案是对RFA-HL-13-027：“筛选基因组命中的功能性试验”（R21/R33）的回应。在过去的15年中，超过40个遗传位点已被确定为与孟德尔和多基因遗传的心律失常和/或心脏性猝死的风险增加相关。随着快速临床基因检测和下一代测序技术的发展，仅最高基因座（LQT 1 -3）的突变/变体数量已超过1000个。然而，这些变体中的绝大多数尚未进行功能表型分型。这给临床遗传学家和管理心脏病专家带来了相当大的困境--未被表征的变异/突变是否被视为心律失常的真正风险？这一决定是至关重要的，因为治疗包括终身药物治疗，行为矫正和心脏起搏器植入。此外，这些管理决策延伸到所有其他携带变异的家庭成员。此外，随着快速、可获得和可负担的全外显子组/基因组在临床竞技场中占据其位置，这些临床情况将很快变得越来越普遍。到目前为止，新变体的功能表型分析一直是以零碎的方式进行的，每次报告一个或少数几个变体。这通常发生在从事心脏生物学其他方面的研究人员的副项目中。我们建议功能表型所有的疾病相关的遗传变异和大多数罕见的多态性，已出现在遗传数据库中的基因座最常报告的遗传性心律失常。将在cDNA中产生变体以在人细胞系中表达。将用高通量电生理学系统评估功能表达。随后，将通过生化分析进一步测定表现出功能丧失表型的那些变体的表面运输缺陷。我们将创建一个具有正在进行的和已完成的结果的医学可用数据库，以允许其他研究者提供输入。这一公共资源的建立将提供 临床遗传学家和管理医生是一个宝贵的资源，以充分分析结果，从基因测试的病人。该项目的成功竞争将为该领域的研究人员提供一个框架和资源，并对越来越多的患者的临床管理产生直接影响。因此，我们认为该提案符合RFA-HL- 13-027和NIH/NHLBI的使命的精神。