Defining an Electrophysiological Phenotype of ALS Patient-Derived Motor Neurons

描述(由申请人提供)：本提案描述了Brian Wainger作为一名使用患者来源的运动神经元研究肌萎缩侧索硬化症(ALS)的研究员实现独立的五年计划。这位候选人是马萨诸塞州综合医院的神经科医生。他在电生理学和分子生物学方面有很强的研究背景，以及最近从患者来源的运动神经元中记录的成功经验，他将把这项技术应用于拟议的项目。首席导师克利福德·伍尔夫博士是哈佛医学院神经学和神经生物学教授，也是波士顿儿童医院神经生物学项目的主任。他是神经科学方面的世界专家，曾指导过许多现在担任学术教职的实习生。共同导师Kevin Eggan博士是HHMI早期职业科学家，也是干细胞和再生生物学系以及哈佛干细胞研究所的教授。他是干细胞领域和肌萎缩侧索硬化症研究的领先者。一个由资深科学家组成的咨询委员会能够提供额外的专家指导，其中还包括梅里特·库德科维奇博士和布鲁斯·宾博士。这项研究将在儿童医院进行。肌萎缩侧索硬化症是一种毁灭性的、无法治愈的神经系统疾病，其特征是运动系统中神经元的进行性衰弱和死亡。大多数病例是零星的，但约10%是家族性的。运动神经元的主要功能是收集和整合来自大脑和脊髓的信号，并传输导致肌肉收缩的传出电信号。研究人员假设，研究健康和疾病运动神经元的电学特性将有助于增加对ALS的了解，并为如何治疗这种疾病提供洞察力。利用干细胞技术的最新进展，研究人员从ALS和对照组受试者身上提取了运动神经元，发现家族性ALS患者的运动神经元具有高度兴奋性，这意味着它们容易过于活跃，这种过度兴奋性可能导致ALS运动神经元的死亡。拟议的项目包括评估ALS的一系列遗传性和零星变种的过度兴奋性和运动神经元死亡。该项目将对观察到的兴奋性进行机械性的理解和表征，特别是关于电压门控钾离子通道的作用。通过培训计划，研究人员将获得使用干细胞生物学和单细胞RNA表达分析技术的关键专业知识，以及教授转化医学的关键成分，如生物统计学和试验设计。有了这些技能，申请者将成功完成拟议的项目并过渡到独立，在那里他可以利用这种新的方法，更广泛地使用人类神经元来模拟人类神经疾病。

DESCRIPTION (provided by applicant): This proposal describes a five-year plan for Brian Wainger to achieve independence as an investigator who uses patient-derived motor neurons to study amyotrophic lateral sclerosis (ALS). The candidate is a neurologist at Massachusetts General Hospital. He has a strong research background in electrophysiology and molecular biology, as well as recent successful experience recording from patient-derived motor neurons, a technique that he will apply to the proposed project. Dr. Clifford Woolf, the primary mentor, is Professor of Neurology and Neurobiology at Harvard Medical School and Director of the Neurobiology Program at Children's Hospital Boston. He is a world expert in neuroscience, and has supervised numerous trainees who now hold academic faculty positions. Dr. Kevin Eggan, the co- mentor, is an HHMI Early Career Scientist and Professor in the Department of Stem Cell and Regenerative Biology and the Harvard Stem Cell Institute. He is a leader in the stem cell field and in ALS research. An advisory committee of senior scientists able to provide additional expert guidance includes also Dr. Merit Cudkowicz and Dr. Bruce Bean. The research will be performed at Children's Hospital. ALS is a devastating, untreatable neurological disease characterized by progressive weakness and death of neurons in the motor system. Most cases are sporadic, but about 10% are familial. The primary function of a motor neuron is to collect and integrate signals from the brain and spinal cord and transmit an outgoing electrical signal that results in muscle contraction. The researchers hypothesize that investigating the electrical properties of healthy and diseased motor neurons will help increase understanding of ALS and yield insight into how to treat the disease. Using recent advances in stem-cell technology, the investigators have derived motor neurons from ALS and control subjects and found that motor neurons derived from patients with familial ALS are hyperexcitable, meaning that they are prone to too much activity, and that the hyperexcitability may contribute to motor neuron death in ALS. The proposed project consists of evaluating hyperexcitability and motor neuron death in a range of inherited and sporadic variants of ALS. The project will yield a mechanistic understanding and characterization of the observed hypexcitability, specifically with regard to the role of voltage-gated potassium ion channels. Through the training program, the investigator will acquire critical expertise using techniques in stem cell biology and single cell RNA expression analysis, as well as didactic exposure to key components of translational medicine such as biostatistics and trial design. With these skills, the applicant will successfully complete the proposed project and transition to independence, where he can exploit this novel approach of modeling human neurological disease using human neurons more broadly.