The role of Hedgehog and enteric neural crest cell reprogramming in neuroendocrine differentiation

项目摘要 尽管越来越多的诊断，胃肠胰腺神经内分泌肿瘤（GEP-NETs）仍然很差， 了解，无论是在其分子发病机制和细胞的起源。我最近的工作是用转基因 小鼠首次证实了肠神经嵴源性细胞（ENCC）参与了 Men1基因缺失导致的神经内分泌分化和肿瘤发展。与其在以下方面的作用相一致： 细胞命运模式，Hedgehog信号转导涉及ENCC的神经内分泌重编程。梅宁， 由MEN1基因编码，已显示在胰腺癌小鼠模型中调节Hedgehog信号传导。 NET的发展，但相互作用之间的menin和刺猬还有待研究，在ENCC。这 该提案旨在解决ENCC中menin的缺失是否会驱动神经内分泌细胞命运的获得， 谱系承诺被Hedgehog信号积极调节。我的目标是定义细胞 MEN1 GEP-NET的起源，并破译Hedgehog在驱动神经内分泌细胞中的作用 模式化我将联合收割机结合我使用新型转基因小鼠模型和离体组织培养的经验， 采用最先进的单细胞和空间转录组分析技术来确定ENCC的贡献 神经内分泌分化和GEP-NET发育。我假设， menin和Hedgehog驱动ENCC重编程并产生增生的神经内分泌细胞 有致瘤潜力本研究计划将确定MEN1相关的GEP-NET是否起源于 从重编程的ENCC群体（目的1）和破译刺猬信号在神经内分泌 MEN1 GEP-NET的重编程（Aim 2）。在指导K01奖励期间，我将与我的 主要导师Juanita Merchant博士和共同导师Megha Padi博士，胃肠道领域的杰出专家 生物学和单细胞分析分别，以发展技能，完成我的研究和职业生涯 目标.我的职业发展计划将促进我成为一个有生产力的独立的目标 研究人员通过结合严格的教学培训和正式的指导下的教师团队谁带来 在细胞和分子生物学、胃肠道生理学、大型基因组数据分析方面的专业知识， 三维类器官系统最后，职业发展资源的广度和广泛的共享研究 亚利桑那大学的设施使它成为我进行拟议研究的理想环境 计划并实现我的职业目标。通过利用创新的GEP-NET模型和尖端的 测序方法，这个奖项将使我能够建立一个国家的最先进的研究计划与长期的- 术语的目标是定义控制肠神经内分泌细胞命运的细胞信号。

PROJECT SUMMARY Despite increasing diagnoses, gastroenteropancreatic neuroendocrine tumors (GEP-NETs) remain poorly understood, both in relation to their molecular pathogenesis and cells-of-origin. My recent work using transgenic mice demonstrated, for the first time, the involvement of enteric neural crest-derived cells (ENCCs) in neuroendocrine differentiation and tumor development driven by Men1 gene deletion. Consistent with its role in cell fate patterning, Hedgehog signaling was implicated in neuroendocrine reprogramming of ENCCs. Menin, encoded by the MEN1 gene, has been shown to regulate Hedgehog signaling in mouse models of pancreatic NET development, however reciprocity between menin and Hedgehog has yet to be studied in ENCCs. This proposal aims to address whether loss of menin in ENCCs drives acquisition of the neuroendocrine cell fate with lineage commitment being actively modulated by Hedgehog signaling. My goals are to define the cells-of- origin for MEN1 GEP-NETs and to decipher the role of Hedgehog in driving neuroendocrine cell patterning. I will combine my experience using novel transgenic mouse models and ex vivo tissue culture techniques with state-of-the-art single cell and spatial transcriptome profiling to define the contribution of ENCCs to neuroendocrine differentiation and GEP-NET development. I hypothesize that reciprocal signaling by menin and Hedgehog drives ENCC reprogramming and gives rise to hyperplastic neuroendocrine cells with tumorigenic potential. This Research Plan will determine whether MEN1-associated GEP-NETs originate from reprogrammed ENCC populations (Aim 1) and decipher the role of Hedgehog signaling in neuroendocrine reprogramming of MEN1 GEP-NETs (Aim 2). During the mentored K01 award period, I will work closely with my primary mentor Dr. Juanita Merchant and co-mentor Dr. Megha Padi, distinguished experts in gastrointestinal biology and single cell analysis respectively, to develop the skillset to accomplish my research and career objectives. My Career Development Plan will facilitate my goal of becoming a productive independent investigator by combining rigorous didactic training and formal mentorship under a team of faculty who bring established expertise in cell and molecular biology, gastrointestinal physiology, large genomic data analysis, and 3-D organoid systems. Finally, the breadth of career development resources and extensive shared research facilities at the University of Arizona make it an ideal environment for me to carry out the proposed research project and achieve my career objectives. By leveraging innovative GEP-NET models and cutting-edge sequencing methods, this award will enable me to establish a state-of-the-art research program with the long- term goal of defining the cellular signals that govern enteric neuroendocrine cell fate.