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

项目摘要/摘要 这份NIH K08建议书描述了最初的五年培训计划，并延长了一年的职业生涯 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研究员 (BWH)和波士顿儿童医院(BCH)，并将开始一项研究 该计划旨在为面向神经疾病的研究进行独立的学术生涯培训。 在这个培训项目中，Rose博士将获得深入研究眼运动神经元(OMN)的经验 发展和轴突靶向，解释单细胞RNA测序，基因调控分析 网络、组织清理和先进的成像方法以及基因编辑技术。他最初的导师 在BCH，伊丽莎白·恩格尔博士(BCH的神经学教授和HHMI研究员)是脑干领域的领导者 OMN发育和涉及这些运动神经元的先天性颅神经紊乱(CCDD)。 随着Rose博士转到加州大学欧文分校(UCI)，加州大学阿尔伯特·拉斯帕达医学博士和UCI博士将 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。 这两家公司都在指导学员在生物医学研究领域取得成功方面建立了良好的记录。 除了他的导师，他还组建了一组在这些领域具有互补专业知识的合作者 以及UCI高级教员咨询委员会，该委员会在指导医生- 科学家通过向独立的过渡。 提出的研究计划的主要科学目标是研究正常和异常的OMN规范 以及轴突生长和对健康和疾病的指导。罗斯博士提供了一张遗传基因的初步图谱 脑干运动神经元的差异可能成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，然后将这些标记应用于对OMN的研究 CCDD。这一建议的中心假设是OMN之间基因表达的差异 在不同的CCDD中，亚群可能倾向于某些亚型经历神经功能障碍。 单细胞rna-seq将在三个主要的动眼神经上进行：动眼神经、滑车神经和外展神经。 脑干核团以发现每个亚群的标记。与ATAC-seq结合，转录 将对OMN规范的监管网络进行调查。每个OMN亚群的唯一标记将 然后被用来研究正常的OMN轴突向不同肌肉靶点的投射，以及用它们来解剖 不同人群对观察到的刻板和病理性异常神经支配的贡献 CCDDS。这些研究对更广泛的神经元规范和分化领域以及 对轴突的生长和引导。

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.