Mapping Brainstem Motor Neuron Subtypes and Genetic Pathways Involved in their Differential Susceptibility to Disease

项目总结/摘要 该NIH K 08提案描述了一个为期五年的初始培训计划，其中包括一年的职业扩展 学术神经病理学和发育神经遗传学的发展。马修·罗斯博士已经完成了 布里格姆妇女医院解剖病理学临床住院医师和神经病理学奖学金 (BWH)和波士顿儿童医院（BCH）在哈佛医学院（HMS），并将着手研究 该计划旨在培养一个独立的学术生涯在神经疾病为导向的研究。 在这个培训项目中，Rose博士将获得眼运动神经元（OMN）研究的深入经验 发育和轴突靶向，单细胞RNA测序的解释，基因调控分析， 网络，组织清除和先进的成像方法，以及基因编辑技术。他最初的导师 在BCH，Elizabeth Engle博士（BCH神经学教授和HHMI研究员）是脑干研究的领导者 OMN发育和涉及这些运动神经元的先天性颅神经支配障碍（CCDD）。 随着博士罗斯的移动到加州大学欧文分校（UCI），阿尔伯特拉斯帕达，医学博士，博士在UCI将 成为罗斯博士的导师，恩格尔博士将继续担任他的共同导师。 两者都建立了指导受训人员在生物医学调查方面取得成功的记录。 除了他的导师，他还召集了一批在这些领域具有互补专长的合作者， 学科，并在UCI的高级教师咨询委员会与指导医生的丰富经验- 科学家向独立过渡。 研究计划的主要科学目标是研究正常和异常OMN规范 以及轴突的生长和健康疾病的指导。罗斯博士提供了一个初步的遗传图谱， 脑干运动神经元之间的差异，可作为进一步研究所有运动神经元的基础 疾病这些初步数据将作为进一步研究OMN发展的基础 通过发现每个OMN亚群的标记物，然后将这些标记物应用于OMN亚群的研究， 的CCD。该建议的中心假设是，OMN中基因表达的差异 亚群可能使某些亚型在不同的CCDD中易发生神经支配障碍。 单细胞RNA-seq将在三个主要的OMN上进行：眼神经、眼轮匝肌和外展神经 脑干细胞核，以发现每个亚群的标记。与ATAC-seq组合， 将研究OMN规范的监管网络。每个OMN亚群的独特标志物将 然后用于研究正常的OMN轴突投射到不同的肌肉目标，以及使用它们来解剖 不同人群的贡献，刻板和病理异常神经支配中观察到的， CCD。这些研究对神经元特化和分化的更广泛领域具有重要意义， 轴突的生长和引导

PROJECT SUMMARY/ABSTRACT This NIH K08 proposal describes an initial five-year training program with one year extension for career development in academic neuropathology and developmental neurogenetics. Dr. Matthew Rose has completed clinical residency in Anatomic Pathology and fellowship in Neuropathology at Brigham and Women’s Hospital (BWH) and Boston Children’s Hospital (BCH) at Harvard Medical School (HMS) and will embark on a research program designed to train for an independent academic career in neurologic disease-oriented research. In this training program, Dr. Rose will acquire in-depth experience in the study of ocular motor neuron (OMN) development and axon targeting, interpretation of single cell RNA-sequencing, analysis of gene regulatory networks, tissue clearing and advanced imaging approaches, and gene-editing technologies. His initial mentor at BCH, Dr. Elizabeth Engle (a Professor of Neurology and HHMI Investigator at BCH), is a leader in brainstem OMN development and congenital cranial dysinnervation disorders (CCDDs) that involve these motor neurons. With Dr. Rose’s move to the University of California, Irvine (UCI), Albert La Spada, MD, PhD at UCI will become Dr. Rose’s mentor, and Dr. Engle will continue as his co-mentor. Both have established track records for mentoring trainees to successful careers in biomedical investigation. In addition to his mentor, he has assembled a group of collaborators with complementary expertise in these disciplines, and an Advisory Committee of senior faculty at UCI with extensive experience in guiding physician- scientists through the transition to independence. The primary scientific goal of the proposed research plan is to study normal and abnormal OMN specification and axonal growth and guidance in health and disease. Dr. Rose provides an initial map of the genetic differences among brainstem motor neurons that may serve as a foundation for further studies of all motor neuron diseases. These preliminary data will be used as a foundation to further study the development of the OMN subnuclei by discovering markers of each OMN subpopulation, and then applying those markers to the study of the CCDDs. The central hypothesis of this proposal is that the differences in gene expression among OMN subpopulations may predispose certain subtypes to undergo dysinnervation in different CCDDs. Single cell RNA-seq will be performed on the three primary OMNs: the oculomotor, trochlear, and abducens brainstem nuclei to discover markers of each subpopulation. In combination with ATAC-seq, the transcriptional regulatory networks for OMN specification will be investigated. Unique markers of each OMN subpopulation will then be used to study normal OMN axon projections to distinct muscle targets, as well as using them to dissect the contributions of different populations to the stereotypic and pathologic aberrant innervations observed in the CCDDs. These studies have significance to the broader fields of neuronal specification and differentiation and to axonal growth and guidance.