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

项目总结 尽管胃肠胰脏神经内分泌肿瘤(GEP-Net)的诊断越来越多，但仍很差。 了解它们与其分子发病机制和起源细胞的关系。我最近的工作是使用转基因 小鼠首次证实肠神经峰源性细胞(ENCCs)参与了 MEN1基因缺失对神经内分泌分化和肿瘤发生的影响与其在以下方面的角色一致 细胞命运模式、Hedgehog信号与ENCC的神经内分泌重编程有关。梅宁 由MEN1基因编码，已被证明在胰腺癌小鼠模型中调节Hedgehog信号。 Net发育，然而，在ENCC中，Menin和Hedgehog之间的相互作用尚未被研究。这 该提案旨在解决ENCCs中薄荷素的丢失是否会导致神经内分泌细胞命运的获得 血统承诺受到刺猬信号的积极调节。我的目标是定义细胞-- MEN1 GEP-Net的起源及Hedgehog在神经内分泌细胞驱动中的作用 图案化。我将把我使用新型转基因小鼠模型和体外组织培养的经验结合起来 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网络发育。我假设互惠的信号通过 梅宁和Hedgehog驱动ENCC重新编程并产生增生性神经内分泌细胞 具有致癌潜力。该研究计划将确定与MEN1相关的GEP-Net是否起源于 并破译Hedgehog信号在神经内分泌中的作用 重新编制MEN1全球环境保护方案网(目标2)。在导师K01颁奖期间，我将与我的 主要导师Juanita Merchant博士和共同导师Megha Padi博士，杰出的胃肠道专家 生物学和单细胞分析，以发展完成我的研究和职业生涯的技能 目标。我的职业发展计划将促进我成为一名富有成效的独立人士的目标 调查员通过将严格的教学培训和正式的指导相结合，在一个教职员工团队的带领下 在细胞和分子生物学、胃肠生理学、大型基因组数据分析以及 3-D有机体系统。最后，广泛的职业发展资源和广泛的共享研究 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 规划并实现我的职业目标。通过利用创新的GEP-Net模式和尖端 测序方法，这个奖项将使我能够建立一个最先进的研究计划，与长期- 定义支配肠道神经内分泌细胞命运的细胞信号的术语目标。

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.