Transcription factors mediate postnatal neurogenesis from enteric glial cells

项目总结 肠道神经系统(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，以及与免疫系统的相互作用。因此，ENS的调节失调 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)。IBDS与肠道神经元密度增加和肠道激活有关 神经胶质细胞，现有证据表明，这些效应直接导致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。 来自我们小组和其他实验室的最新论文表明，一些神经胶质细胞 对炎症做出反应，改变命运成为神经元。因此，ENS胶质细胞包含神经元的一个子集 祖细胞和炎症信号触发这个群体产生新的神经元。然而，它并不是 明确神经胶质细胞如何改变其转录程序以产生神经元，以及神经胶质细胞的哪些亚群 参与这一过程的人还不清楚。这个项目的目标是测试特定转录的假设 已知在出生前神经发生中起作用的因素在ENS中诱导出生后神经发生 发炎。PHOX2B是一种转录因子，在ENS和中枢神经系统中都有重要作用 神经发生，初步数据表明，它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得 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 基因敲除小鼠模型。该项目的完成将显著提高我们对肠胶质细胞的了解。 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有可能被利用来治疗再生缺失 神经元。除了建议的研究外，该奖学金还将提供计划、执行和分析方面的培训 来自利用RNA测序技术培育小鼠以产生组织特异性的实验数据 条件性基因敲除模型，用于培养和操纵原代肠道神经胶质细胞，以有效地沟通 以书面和口头形式提供科学数据，并管理和领导科学项目。在完成 奖学金，我将准备开始我自己的职业生涯，作为一名独立的，由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.