NEXT-GENERATION SEQUENCING APPROACHES FOR NOVEL GENE DISCOVERY IN ALS

用于 ALS 新基因发现的下一代测序方法

基本信息

批准号：
8481601
负责人：
Matthew Harms
金额：
$ 15.74万
依托单位：
WASHINGTON UNIVERSITY
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：
2011-07-01 至 2016-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8481601
关键词：
Age of Onset Algorithms Amyotrophic Lateral Sclerosis Animal Model Bioinformatics Biological Assay Biometry Brain Stem Candidate Disease Gene Cessation of life Clinical Code Collaborations Communities Coupled Cytoplasm DNA Data Dementia Development Plans Disease Disease Management Exons Familial Amyotrophic Lateral Sclerosis Family Future Gene Mutation Genes Genetic Genetic Heterogeneity Genetic Techniques Genome Genomics Genotype Goals Human Human Genetics Human Genome Incidence Individual Inherited Institution Introns Laboratories Lead Light Mentors Methods Minority Modeling Molecular Molecular Genetics Motor Neurons Mutation Nerve Degeneration Neurodegenerative Disorders Neurologic Neurologist Neuromuscular Diseases Neuropharmacology Nuclear Paralysed Pathogenesis Pathogenicity Pathway interactions Patients Penetrance RNA Processing RNA-Binding Proteins Research Research Personnel Research Project Grants Research Proposals Respiratory Failure Sampling Spinal Cord Symptoms Techniques Technology Testing Training Universities Variant Washington Work addiction base career career development cohort disease-causing mutation effective therapy exome exome sequencing experience follow-up gene discovery gene function genetic linkage analysis genetic pedigree innovation insight motor neuron degeneration neurodevelopment neurogenetics next generation sequencing novel novel strategies palliation proband protein TDP-43 public health relevance repository respiratory segregation therapeutic development tissue culture tool

项目摘要

DESCRIPTION (provided by applicant): The candidate is a clinical neurologist (MD) with a career goal of investigating the genetic and molecular basis of human neuromuscular diseases, focusing primarily on amyotrophic lateral sclerosis (ALS). The candidate has significant prior laboratory experience, with a track record of successful research projects in neuropharmacology, neurodevelopment, and most recently, neurogenetics. In order to further prepare for successful independent research, the candidate's career development plan includes graduate-level courses in biostatistics, bioinformatics, and genomics, as well as seminars and intensive courses on next-generation sequencing and related methods. The proposed development plan and scientific training will take place at Washington University in St. Louis, an institution with particular strengths in both genetics/genomics and the study of ALS. The neurogenetics community is robust and will provide the candidate with important intellectual assistance and collaborations. The scientific training will be jointly mentored by Dr. Alison Goate, whose lab focuses on the genetics of dementia and addiction, and Dr. Robert Baloh, whose lab studies the molecular basis of neurodegeneration in ALS using tissue culture and animal models. Dr. Goate's expertise in human genetics and sequencing methods coupled with Dr. Baloh's experience in modeling human neuromuscular diseases will provide the candidate with the set of research tools needed to succeed as an independent investigator studying the genetic and molecular basis of neurologic diseases. The candidate's research proposal aims to identify novel genes causing ALS, an untreatable neurodegenerative disease producing loss of motor neurons, progressive paralysis, and death from respiratory failure. Although the majority of ALS cases are sporadic, the genes responsible in familial cases have provided important insights into all forms of the disease. However, known ALS genes still only explain 25% of familial cases. To identify additional familial ALS disease genes, the candidate will: 1) Screen a large cohort of familial and sporadic ALS patients for mutations in known ALS genes. This screen will be performed using pooled-sample sequencing, an innovative method made possible by next-generation sequencing capabilities pioneered by one of our key collaborators. Novel disease-causing mutations will be followed up with phenotypic analysis. This screen will also identify a cohort of ALS families without mutations in any known ALS disease gene to be studied with whole-exome sequencing. 2) Familial cases of ALS without mutations in known genes will then undergo whole-exome analysis, a next- generation sequencing method that allows all coding regions in the human genome to be sequenced simultaneously for variations. To narrow the number of candidate pathogenic mutations, a host of bioinformatic techniques will be used. Candidate ALS genes identified will then be screened for additional mutations in other ALS patients using the same pooled-sample sequencing strategy as in (1). The identification of novel ALS disease genes will pave the way for future studies aimed at understanding the molecular mechanism by which these mutations lead to neurodegeneration, using tissue culture and animal modeling. This work will provide key insight into the molecular basis of ALS and will immediately impact efforts aimed at therapeutic development for this untreatable disease.

描述（由申请人提供）：候选人是一名临床神经学家（MD），其职业目标是研究人类神经肌肉疾病的遗传和分子基础，主要关注肌萎缩侧索硬化症（ALS）。该候选人之前拥有丰富的实验室经验，在神经药理学、神经发育以及最近的神经遗传学领域拥有成功的研究项目记录。为了进一步为成功的独立研究做好准备，候选人的职业发展计划包括生物统计学、生物信息学和基因组学的研究生水平课程，以及下一代测序和相关方法的研讨会和强化课程。拟议的发展计划和科学培训将在圣路易斯华盛顿大学进行，该机构在遗传学/基因组学和 ALS 研究方面具有独特的优势。神经遗传学界非常强大，将为候选人提供重要的智力帮助和合作。科学培训将由艾莉森·戈特博士和罗伯特·巴洛博士共同指导，艾莉森·戈特博士的实验室专注于痴呆和成瘾的遗传学，罗伯特·巴洛博士的实验室利用组织培养和动物模型研究肌萎缩侧索硬化症神经退行性变的分子基础。 Goate 博士在人类遗传学和测序方法方面的专业知识，加上 Baloh 博士在人类神经肌肉疾病建模方面的经验，将为候选人提供一套成功作为独立研究者研究神经系统疾病的遗传和分子基础所需的研究工具。该候选人的研究计划旨在识别导致 ALS 的新基因，ALS 是一种无法治疗的神经退行性疾病，会导致运动神经元丧失、进行性瘫痪和呼吸衰竭死亡。尽管大多数 ALS 病例是散发性的，但家族病例中的相关基因为了解该疾病的所有形式提供了重要的见解。然而，已知的 ALS 基因仍然只能解释 25% 的家族性病例。为了识别其他家族性 ALS 疾病基因，候选人将： 1) 对一大群家族性和散发性 ALS 患者进行已知 ALS 基因突变的筛查。该筛选将使用混合样本测序进行，这是一种创新方法，由我们的一位主要合作者首创的下一代测序功能实现。新的致病突变将进行表型分析。该筛查还将鉴定出一组没有任何已知 ALS 疾病基因突变的 ALS 家族，以便通过全外显子组测序进行研究。 2) 已知基因没有突变的 ALS 家族病例将接受全外显子组分析，这是一种下一代测序方法，允许对人类基因组中的所有编码区域同时进行变异测序。为了缩小候选致病突变的数量，将使用大量生物信息学技术。然后，将使用与 (1) 中相同的合并样本测序策略，筛选其他 ALS 患者中确定的候选 ALS 基因是否存在其他突变。新的 ALS 疾病基因的鉴定将为未来的研究铺平道路，旨在利用组织培养和动物模型了解这些突变导致神经退行性变的分子机制。这项工作将为 ALS 的分子基础提供重要的见解，并将立即影响针对这种无法治疗的疾病的治疗开发工作。