Exome re-sequencing candidate loci for familial essential tremor

家族性特发性震颤候选位点的外显子组重测序

基本信息

批准号：
8164921
负责人：
BARRY E KOSOFSKY
金额：
$ 25.35万
依托单位：
WEILL MEDICAL COLL OF CORNELL UNIV
依托单位国家：
美国
项目类别：
财政年份：
2011
资助国家：
美国
起止时间：
2011-05-15 至 2013-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8164921
关键词：
2p24 6p23 Address Affect Aging Alleles American Ataxia Base Pairing Binding Proteins Bioinformatics Candidate Disease Gene Chromosomes Classification Complex Computer software DNA DNA Resequencing DNA Sequence Databases Development Diagnostic Dideoxy Chain Termination DNA Sequencing Disease Dominant Genetic Conditions Dystonia Enhancers Essential Tremor European Exons Family Feline Leukemia Virus Future Gene Mutation Genes Genetic Genetic Counseling Genetic Predisposition to Disease Genetic screening method Genome Haplotypes Hematopoietic High Prevalence Human In Vitro Individual Inheritance Patterns Inherited Intercistronic Region Knowledge Korea Koreans Lead Length Libraries Light Link Lod Score Manuscripts Methodology Missense Mutation Molecular Movement Disorders Mutation Neurobiology Nucleic Acid Regulatory Sequences Nucleotides Outcome Parkinson Disease Penetrance Phase Phenocopy Phenotype Polymorphism Analysis Population Population Database Predisposition Prevalence Process Proteins RNA Splicing Reporting Research Research Personnel Retinitis Pigmentosa Sample Size Screening procedure Singapore Single-Stranded Conformational Polymorphism Site Solid Subgroup Techniques Technology Therapeutic Intervention Transcript Tremor United States Variant base clinical practice density design diagnostic accuracy dopamine D3 receptor dorsal column effective therapy exome genetic association genetic linkage genetic variant genome-wide linkage improved in vivo Model instrumentation mutant nervous system disorder novel novel strategies outcome forecast receptor research study transmission process

项目摘要

DESCRIPTION (provided by applicant): Essential tremor (ET) is one of the most common neurological disorders in humans with a prevalence reaching almost four percent. ET is inherited as a dominant trait with incomplete penetrance in most familial cases but complex multi-genic transmission is possible. Three genetic susceptibility loci for familial ET exist on chromosomes 3p13.1 (ETM1), 2p24 (ETM2), and 6p23 (ETM3), but the causal genes have not been identified in the families linked to these loci. Polymorphic ETM1 loci in 16 Icelandic families and a variant in the dopamine receptor D3 gene in 23 French families are associated with an ET phenotype. ETM2 loci are linked to a disease allele in four American families. Further studies suggest a tight allelic association between ETM2 and the ET phenotype in populations from the United States, Singapore, and Korea. Two North American families are linked to ETM3 loci with the largest family showing a mixed phenotype that includes dystonia in addition to ET. An unclear pattern of inheritance due to the presence of phenocopies, incomplete penetrance, and the high prevalence of ET, hinder the search for causal genes. Advances in solid- phase microarrays, and instrumentation have partially alleviated these barriers by permitting large scale DNA sequencing. Advances in genome bioinformatics and the availability of validated normative population databases (e.g. 1000 Genomes SNP database, dbSNP, and HapMap) provide the capability to filter genetic variants from putative mutations. By using this technology, we recently identified missense mutations in a novel gene, feline leukemia virus subgroup C receptor 1 gene, as the cause of the Mendelian disorder, posterior column ataxia and retinitis pigmentosa, in three unrelated families by high density sequencing of the 4.2 megabase (Mb) candidate region. This discovery confirms our expertise in all of the specific technologies required to carry out the proposed genetic studies and provides proof of principle for Specific Aim 1 to identify the genes that cause ET in two large, informative families linked to the ETM2 and ETM3 loci using high- throughput resequencing technology. We have designed a bait tiling library using 120 base-pair bait lengths for the 24.6 Mb ETM2 (48,054 baits) and the 14.4 Mb ETM3 (30,796 baits) loci to include all regulatory regions, exons, splice sites, enhancer, and conserved intergenic regions. A power analysis estimates that the sample size required to detect mutations with 95% confidence are 10 affected individuals with a disease haplotype and 10 unaffected individuals without the disease haplotype from each family linked to the ETM2 and ETM3 loci. Sanger sequencing will reconfirm the ETM2 and ETM3 mutations identified in the 20 affected individuals from the two families. These candidate genes will be Sanger sequenced in 73 additional, unrelated families with ET to identify other ETM2 and ETM3 mutations. Future experiments using in vitro and in vivo models will analyze the functional consequences of ET gene mutations during aging and development. PUBLIC HEALTH RELEVANCE: The identification of ET genes will improve diagnostic accuracy, refine the classification of tremor, and is a pivotal step toward finding effective treatments. Studying the untoward effects of mutant ET genes will increase our knowledge of ET and other movement disorders such as Parkinson disease.

描述（由申请人提供）：基本震颤（ET）是人类中最常见的神经系统疾病之一，患病率达到近4％。 ET在大多数家族案例中被遗传为具有不完全渗透率的主要特征，但是复杂的多类型传播是可能的。染色体3P13.1（ETM1），2P24（ETM2）和6P23（ETM3）上存在三个遗传易感性基因座，但在与这些基因座相关的家族中尚未鉴定出因果基因。 16个冰岛家族中的多态性ETM1基因座和23个法国家族中多巴胺受体D3基因的变体与ET表型相关。 ETM2基因座与四个美国家庭中的疾病等位基因有关。进一步的研究表明，在美国，新加坡和韩国人群中，ETM2与ET表型之间存在紧密的等位基因关联。两个北美家庭与ETM3基因座有关，其中最大的家族表现出混合表型，除了ET外，还包括肌张力障碍。由于表型的存在，不完全的外观和ET的高流行而导致的遗传模式不明显，阻碍了对因果基因的搜索。固相微阵列和仪器的进步通过允许大规模的DNA测序部分缓解了这些障碍。基因组生物信息学的进步以及经过验证的规范总体数据库的可用性（例如1000个基因组SNP数据库，DBSNP和HAPMAP）提供了从推定突变中过滤遗传变异的能力。通过使用这项技术，我们最近确定了一种新型基因中的错义突变，猫白血病病毒亚组C受体1基因是Mendelian疾病，后柱共济失调和色素性视网膜炎的原因，在三个不相关的家族中，由4.2兆巴群（MB）候选区域的高密度测序中的三个无关家族中。这一发现证实了我们在进行提出的遗传研究所需的所有特定技术中的专业知识，并为特定目标1提供了原理证明，以识别使用高吞吐量重新设备技术与ETM2和ETM3 Loci相关的两个大型，信息丰富的家族中引起ET的基因。我们已经使用120个碱基对诱饵长度设计了一个诱饵瓷砖库，用于24.6 MB ETM2（48,054个诱饵）和14.4 MB ETM3（30,796个诱饵）基因座，包括所有调节区域，外显子，夹杂物，增强器，增强器和保守的地间区域。功率分析估计，检测95％置信度突变所需的样本量是10个受疾病单倍型的受影响的个体和10个未受影响的个体，而没有疾病单倍型，来自与ETM2和ETM3基因座相关的每个家庭。 Sanger测序将重新确认来自两个家庭的20个受影响个体的ETM2和ETM3突变。这些候选基因将在73个具有ET的无关家族中进行桑格测序，以鉴定其他ETM2和ETM3突变。未来使用体外和体内模型的实验将分析ET基因突变在衰老和发育过程中的功能后果。公共卫生相关性：ET基因的识别将提高诊断准确性，完善震颤的分类，并且是寻找有效治疗方法的关键步骤。研究突变ET基因的不良影响将增加我们对ET和其他运动障碍（例如帕金森病）的了解。