Defining an Electrophysiological Phenotype of ALS Patient-Derived Motor Neurons

描述（由申请人提供）：该提案描述了Brian winger作为一名使用患者源性运动神经元研究肌萎缩性侧索硬化症（ALS）的研究者实现独立性的五年计划。候选人是马萨诸塞州总医院的神经科医生。他在电生理学和分子生物学方面有很强的研究背景，以及最近成功记录来自患者的运动神经元的经验，他将把这项技术应用到拟议的项目中。主要导师Clifford Woolf博士是哈佛医学院神经病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主任。他是神经科学领域的世界级专家，并指导了许多现在担任学术教职的学员。共同导师Kevin Eggan博士是HHMI早期职业科学家和哈佛干细胞研究所干细胞和再生生物学部门的教授。他是干细胞领域和ALS研究领域的领军人物。能够提供额外专家指导的资深科学家咨询委员会还包括Merit Cudkowicz博士和Bruce Bean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的神经元逐渐衰弱和死亡。大多数病例是散发的，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，并向外传递导致肌肉收缩的电信号。研究人员假设，调查健康和患病运动神经元的电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见解。利用干细胞技术的最新进展，研究人员从ALS和对照受试者中提取了运动神经元，发现来自家族性ALS患者的运动神经元是过度兴奋的，这意味着它们容易过度活动，这种过度兴奋性可能导致ALS患者的运动神经元死亡。该计划包括评估一系列遗传性和散发性ALS变体的过度兴奋性和运动神经元死亡。该项目将产生对观察到的低兴奋性的机制理解和表征，特别是关于电压门控钾离子通道的作用。通过培训计划，研究者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，以及生物统计学和试验设计等转化医学的关键组成部分的教学。有了这些技能，申请人将成功完成拟议的项目并过渡到独立，在那里他可以更广泛地利用这种利用人类神经元建模人类神经系统疾病的新方法。

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.