Transcription factors mediate postnatal neurogenesis from enteric glial cells

项目摘要 肠神经系统（ENS）通过调节肠道运动在胃肠道生物学中起着至关重要的作用， 分泌和吸收，以及与免疫系统相互作用。ENS失调， 导致许多疾病，包括炎症性肠病（IBD）、溃疡性结肠炎（UC）和 克罗恩病（CD）。IBD与肠神经元密度增加和肠上皮细胞活化有关。 神经胶质细胞，现有的证据表明，这些影响直接有助于IBD的病理生物学。 因此，了解ENS组分在UC和CD中的作用对于开发靶向治疗至关重要。 我们小组和其他实验室最近的论文都表明，一些ENS神经胶质细胞 在炎症反应中转变命运成为神经元。ENS神经胶质因此含有神经元的子集， 祖细胞和炎症信号触发该群体产生新的神经元。但不 清楚神经胶质细胞如何改变它们的转录程序以产生神经元，以及神经胶质细胞的亚群是什么。 参与这一过程的是未知的。这个项目的目标是检验特异性转录 在产前神经发生中具有已知作用的因子诱导ENS中的出生后神经发生， 炎症Phox2b是一种在ENS和中枢神经系统中具有重要作用的转录因子 神经发生，并且初步数据显示其诱导培养的神经胶质细胞采用神经元表型。我 假设结肠炎症将触发Phox2b介导的神经元命运获取， 肠神经胶质细胞。为了验证这一假设，我将使用单细胞RNA测序和条件Phox2b 敲除小鼠模型。这个项目的完成将大大提高我们对肠神经胶质细胞的理解， IBD的病理生理学，并将揭示可能代表治疗靶点的分子途径。这 这项工作还将揭示神经胶质细胞亚群的潜力，利用治疗再生失踪 神经元除了拟议的研究，该奖学金将提供培训，以计划，执行和分析 利用RNA测序技术的实验数据，以繁殖小鼠， 条件敲除模型，培养和操纵原代肠神经胶质细胞，有效地沟通 科学数据的书面和口头，并管理和领导一个科学项目。完成后 奖学金，我将准备开始我自己的职业生涯作为一个独立的，NIH资助的科学家研究 胃肠生物学

PROJECT SUMMARY The enteric nervous system (ENS) plays a vital role in gastrointestinal biology by regulating gut motility, secretion, and absorption, and by interacting with the immune system. Dysregulation of the ENS thus contributes to many diseases, including the inflammatory bowel diseases (IBDs) ulcerative colitis (UC) and Crohn’s disease (CD). IBDs are associated with increased density of enteric neurons and activation of enteric glial cells, and existing evidence suggests that these effects directly contribute to the pathobiology of IBDs. Understanding the role of ENS components in UC and CD is thus critical for developing targeted therapies. Recent papers both from our group and from other laboratories have demonstrated that some ENS glial cells switch fates to become neurons in response to inflammation. The ENS glia thus contain a subset of neuronal progenitor cells, and inflammatory signals trigger this population to generate new neurons. However, it is not clear how glial cells alter their transcriptional program to generate neurons, and what subgroups of glia are involved in this process is unknown. The goal of this project is to test the hypothesis that specific transcription factors with known roles in prenatal neurogenesis induce postnatal neurogenesis in the ENS in response to inflammation. Phox2b is a transcription factor with critical roles in both ENS and central nervous system neurogenesis, and preliminary data shows that it induces cultured glia to adopt a neuronal phenotype. I hypothesize that colonic inflammation will trigger Phox2b-mediated neuronal fate acquisition in a subset of enteric glial cells. To test this hypothesis, I will use single cell RNA sequencing and a conditional Phox2b knockout mouse model. Completion of this project will significantly enhance our understanding of enteric glia in the pathophysiology of IBDs, and will reveal molecular pathways that may represent therapeutic targets. This work will also reveal glial subpopulations with potential to be harnessed therapeutically to regenerate missing neurons. In addition to the proposed research, this fellowship will provide training to plan, execute, and analyze data from experiments utilizing RNA sequencing techniques, to breed mice to generate tissue-specific conditional knockout models, to culture and manipulate primary enteric glial cells, to effectively communicate scientific data in writing and orally, and to manage and lead a scientific project. Upon completion of the fellowship, I will be prepared to launch my own career as an independent, NIH-funded scientist studying gastrointestinal biology.