Gene regulatory mechanisms that assign and maintain motor neuron terminal differe

概述：运动神经元（MN）功能或存活缺陷会导致严重的人类疾病，如肌萎缩侧索硬化症（ALS）和脊髓性肌萎缩症（SMA）。MN疾病没有有效的治疗方法，部分原因是我们对MN发展的最后步骤，即MN末端分化基因的诱导了解不足。此外，在成年期维持MN末端分化基因表达（从而确保MN功能）的分子机制在很大程度上是未知的。我们最近首次提供了在发育过程中建立MN末端分化并在成人生活中维持这种分化的调控机制的见解。使用线虫和简单脊索动物作为模型系统，我们发现COE （Collier, Olf, Ebf）家族转录因子分配并维持MN末端分化。在这个职业发展提案中，C在Aim #1中，我将利用我在条件小鼠诱变方面的专业知识来检验Ebf2 （COE家族成员）是脊髓MNs最终分化所必需的假设。Aim #2的目标是利用芯片测序和微阵列技术在小鼠胚胎干细胞（mESC）衍生的MN上鉴定脊椎动物中新的MN末端分化标记物。这是MN领域一个备受期待的目标，因为目前只有乙酰胆碱（ACh）途径基因作为MN末端分化的真正标记。在Aim #3中，我将致力于揭示秀丽隐杆线虫特定MN亚型的终端分化所需的其他进化保守转录因子。从这些目标中获得的知识将有助于我们了解MN末端分化是如何在发育过程中建立并在整个生命中维持的，并可能为MN疾病的病因、诊断或治疗提供新的切入点。如果获得资助，该职业发展奖（CDA）将为我提供机会扩展我在秀丽隐杆线虫遗传学方面的背景，发展脊椎动物神经生物学方面的额外专业知识，获得芯片测序和生物信息学工具使用方面的必要培训；这些技能对我未来作为独立研究者的成功至关重要。与我的咨询委员会的正式和非正式互动将评估我在拟议目标方面的进展并提供咨询意见。最后，在K99阶段，通过哥伦比亚大学的课程和我的顾问委员会和导师的建议，我将获得对我作为一名学者取得长期成功至关重要的专业技能（如指导、研究报告、时间管理）。我提出的学习和职业发展计划将确保我实现我的长期目标；在一所重点大学建立一个成功的实验室，并为MN领域做出重大贡献。

DESCRIPTION (provided by candidate): "Gene regulatory mechanisms that assign and maintain motor neuron terminal differentiation" Summary: Defects in motor neuron (MN) function or survival result in severe human conditions such as amyotrophic lateral sclerosis (ALS), and spinal muscular atrophy (SMA). There are no effective treatments for MN disorders in part due to our poor understanding of the last steps of MN development, i.e., the induction of MN terminal differentiation genes. In addition, the molecular mechanisms that maintain the expression of MN terminal differentiation genes during adult life (and thereby ensure MN function) are largely unknown. We have recently provided the first insights on the regulatory mechanisms that establish MN terminal differentiation during development and maintain it in adult life. Using a nematode and a simple chordate as model systems, we showed that the COE (Collier, Olf, Ebf) family of transcription factors assigns and maintains MN terminal differentiation. In this Career Development Proposal, C In Aim #1, I will test the hypothesis that Ebf2 (COE family member) is required for terminal differentiation of spinal MNs using my expertise in conditional mouse mutagenesis. The goal of Aim #2 is to identify novel MN terminal differentiation markers in vertebrates using ChIP-sequencing and microarray technology on mouse embryonic stem cell (mESC)-derived MNs. This is a much-anticipated goal in the MN field as currently only the acetylcholine (ACh) pathway genes serve as bona fide markers of MN terminal differentiation. In Aim #3, I will aim to reveal other evolutionarily conserved transcripton factors that are required for terminal differentiation of specific MN subtypes in C. Elegans. The knowledge gained from these aims will lend to our understanding of how MN terminal differentiation is established during development and maintained throughout life, and may provide novel entry points into the etiology, diagnosis or treatment of MN disorders. If funded, this Career development Award (CDA) would afford me the opportunity to expand my background in C. Elegans genetics, develop additional expertise in vertebrate neurobiology, obtain the necessary training in ChIP-sequencing and the use of bioinformatic tools; skills critical for my future success as an independent researcher. Formal and informal interactions with my Advisory Committee will assess my progress on the proposed Aims and provide advice. Finally, professional skills (e.g. mentoring, research presentation, time management) vital to my long-term success as an academic will be gained during the K99 phase through Columbia University courses and advice by my advisory committee and my mentor. Together, the proposed studies and career development plan will ensure I achieve my long-term goal; to establish a successful laboratory at a major University and make significant contributions to the MN field.