Analysis of tRNA Synthetase Variants in the Undiagnosed Diseases Program

未确诊疾病项目中 tRNA 合成酶变异体的分析

基本信息

批准号：
8679822
负责人：
Anthony Antonellis
金额：
$ 24.07万
依托单位：
UNIVERSITY OF MICHIGAN AT ANN ARBOR
依托单位国家：
美国
项目类别：
财政年份：
2014
资助国家：
美国
起止时间：
2014-09-01 至 2016-07-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8679822
关键词：
Affect Alanine-tRNA Ligase Alleles Amino Acids Amino Acyl-tRNA Synthetases Aminoacylation Aspartate-tRNA Ligase Ataxia Binding Biochemical Biological Assay Biological Models Body Weight decreased Cardiomyopathies Cataloging Catalogs Catalytic Domain Cell Culture Techniques Cell Line Cells Communities Cultured Cells Cytoplasm DNA DNA Sequence Developmental Delay Disorders Disease Encephalopathies Enzymes Evaluation Failure to Thrive Family Family history of Family member Genes Genetic Genome Glycine-tRNA Ligase Hearing Hereditary Disease Human Human Genetics Impaired cognition In Vitro Individual Laboratories Link Microcephaly Molecular Mutation Nerve Degeneration Nuclear Onset of illness Ovarian Parents Pathogenicity Patients Peripheral Nervous System Diseases Phenotype Proteins Research Role Sampling Series Siblings Spastic Paraplegia Supporting Cell Technology Testing Therapeutic Toxic effect Transfer RNA Transfer RNA Aminoacylation Translations Variant Yeast Model System Yeasts Zebrafish base cell growth clinical phenotype clinically relevant cohort disease phenotype disease-causing mutation early onset gain of function genetic pedigree human disease human tissue improved in vivo insight loss of function next generation sequencing novel proband programs public health relevance rare variant research clinical testing therapy development

项目摘要

DESCRIPTION (provided by applicant): Rapid advances in DNA sequencing technologies have allowed comprehensive analysis of patient samples for disease-associated mutations. A major complicating factor is that mutations are often found in small pedigrees or sporadic cases of disease making it difficult to build a strong genetic argument for pathogenicity. In these cases functional analyses are critical for evaluating the pathogenicity of rare variants. Furthermore, functional evaluation can provide insight into the molecular mechanism of disease as well as platforms for developing therapeutics. The Undiagnosed Diseases Program (UDP) is cataloging promising rare variants identified in patients with myriad clinical phenotypes. A subset of these variants reside in genes encoding aminoacyl-tRNA synthetases (ARSs), which are critical enzymes involved in charging tRNA with cognate amino acids. The UDP has identified six ARS variants-two each in glycyl-tRNA synthetase (GARS), alanyl-tRNA synthetase (AARS), and aspartyl-tRNA synthetase (DARS). Our laboratory specializes in evaluating the pathogenicity of ARS variants in vitro and in vivo. Indeed, we identified the first disease-associated ARS mutations in the GARS gene in 2003 and have been continuously studying this class of genes ever since. We propose two specific aims to evaluate the pathogenicity of the six ARS variants and to understand how they cause human genetic disease. First, we will test each ARS variant for a loss-of-function effect in biochemical, yeast complementation, and cellular localization assays. Second, we will determine if each ARS variant is able to exert a dominant, toxic phenotype in zebrafish, similar to other disease-associated ARS mutations. These efforts compose a thorough evaluation of the pathogenic potential of each ARS variant and will provide the basis for developing therapies for affected individuals.

描述（由申请人提供）：DNA 测序技术的快速进步使得能够对患者样本进行与疾病相关的突变的全面分析。一个主要的复杂因素是，突变经常出现在小谱系或零星疾病病例中，因此很难为致病性建立强有力的遗传论据。在这些情况下，功能分析对于评估罕见变异的致病性至关重要。此外，功能评估可以深入了解疾病的分子机制以及开发治疗方法的平台。未诊断疾病计划 (UDP) 正在对在具有多种临床表型的患者中发现的有希望的罕见变异进行分类。这些变体的一个子集存在于编码氨酰基-tRNA 合成酶 (ARS) 的基因中，氨酰基-tRNA 合成酶是参与向 tRNA 添加同源氨基酸的关键酶。 UDP 已鉴定出六种 ARS 变体，其中甘氨酰 tRNA 合成酶 (GARS)、丙氨酰 tRNA 合成酶 (AARS) 和天冬氨酰 tRNA 合成酶 (DARS) 各有两种。我们的实验室专门评估 ARS 变异的体外和体内致病性。事实上，我们于 2003 年在 GARS 基因中发现了第一个与疾病相关的 ARS 突变，并从那时起一直在不断研究此类基因。我们提出了两个具体目标来评估六种 ARS 变体的致病性并了解它们如何导致人类遗传疾病。首先，我们将测试每个 ARS 变体在生化、酵母互补和细胞定位测定中的功能丧失效应。其次，我们将确定每个 ARS 变异是否能够在斑马鱼中发挥显性的毒性表型，类似于其他与疾病相关的 ARS 突变。这些工作对每种 ARS 变体的致病潜力进行了全面评估，并将为开发针对受影响个体的治疗方法提供基础。