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

• 批准号: 9341796
• 负责人: Matthew Harms
• 金额: $ 15.12万
• 依托单位: COLUMBIA UNIVERSITY HEALTH SCIENCES
• 依托单位国家: 美国
• 财政年份: 2016
• 资助国家: 美国
• 起止时间: 2016-09-01 至 2017-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9341796
• 关键词: Next; Generation; Sequencing; Approaches; Novel

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科医生（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，主要集中于肌萎缩性侧向硬化症（ALS）。候选人具有先前的实验室经验，并具有神经药理，神经发育和最近的神经遗传学的成功研究项目的记录。为了进一步准备成功的独立研究，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方面的研究生级课程，以及有关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的华盛顿大学进行，该机构在遗传学/基因组学和ALS研究方面具有特殊优势。神经遗传学社区非常强大，将为候选人提供重要的智力援助和合作。科学培训将由Alison Goate博士共同指导，他的实验室专注于痴呆和成瘾的遗传学，Robert Baloh博士使用组织培养和动物模型研究了ALS中神经变性的分子基础。 Goate博士在人类遗传学和测序方法方面的专业知识以及Baloh博士在建模人类神经肌肉疾病方面的经验将为候选人提供成功的研究工具，作为研究神经疾病的遗传和分子基础所需的成功研究工具。候选人的研究建议旨在鉴定引起ALS的新基因，这是一种无法治疗的神经退行性疾病，导致运动神经元的丧失，进行性瘫痪以及呼吸衰竭死亡。尽管大多数ALS病例都是零星的，但在家族病例中负责的基因为所有形式的疾病提供了重要的见解。但是，已知的ALS基因仍然仅解释25％的家族病例。为了鉴定其他家族性ALS疾病基因，候选人将：1）筛选大量家族性和零星ALS患者在已知ALS基因中的突变。该屏幕将使用合并样本测序进行执行，这是一种创新方法，这是通过我们的一位主要协作者开创的下一代测序能力做出的创新方法。新型致病突变将进行表型分析。该屏幕还将在任何已知的ALS疾病基因中都没有突变来识别一群ALS家族，该基因将通过全外象征进行研究。 2）随后在已知基因中没有突变的ALS的家族病例将经过全外检查分析，这是一种下一代测序方法，允许人类基因组中的所有编码区域同时对变异进行测序。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，使用与（1）中相同的合并样本测序策略，将对其他ALS患者进行其他突变进行筛选，以筛选其他ALS患者的其他突变。新型ALS疾病基因的鉴定将为未来的研究铺平道路，旨在理解这些突变通过组织培养和动物建模的分子机制导致神经变性的方法。这项工作将为ALS的分子基础提供关键的见解，并将立即影响旨在用于这种不可治疗疾病的治疗发展的努力。