Enteroendocrine Progenitor Cell–Enriched miR-7 Regulates Intestinal Epithelial Proliferation in an Xiap-Dependent Manner
Enteroendocrine Progenitor Cell–Enriched miR-7 Regulates Intestinal Epithelial Proliferation in an Xiap-Dependent Manner
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DOI：
10.1016/j.jcmgh.2019.11.001
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发表时间：
2019-11
影响因子：
7.2
通讯作者：
A. Singh;Y. Hung;M. Shanahan;Matt Kanke;Alessandro Bonfini;M. Dame;Mandy Biraud;B. Peck;O. Oyesola;J. Freund;Rebecca L Cubitt;Ennessa G. Curry;L. Gonzalez;G. Bewick;Elia D. Tait-Wojno;N. Kurpios;Shengli Ding;J. Spence;C. M. Dekaney;N. Buchon;P. Sethupathy
中科院分区：
文献类型：
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作者：
A. Singh;Y. Hung;M. Shanahan;Matt Kanke;Alessandro Bonfini;M. Dame;Mandy Biraud;B. Peck;O. Oyesola;J. Freund;Rebecca L Cubitt;Ennessa G. Curry;L. Gonzalez;G. Bewick;Elia D. Tait-Wojno;N. Kurpios;Shengli Ding;J. Spence;C. M. Dekaney;N. Buchon;P. Sethupathy
背景与目的肠内分泌细胞（EEC）是维持肠道内环境稳定的重要细胞系。最近的研究表明，EEC祖细胞有助于肠上皮细胞的生长和更新，但其潜在的机制仍然知之甚少。microRNA是根据探索沿着整个EEC谱系轨迹，相对较少的是知道他们的贡献肠homeostasis.MethodsWe利用无偏测序和八个不同的小鼠模型和分选方法，以确定microRNA丰富沿着EEC谱系轨迹。我们进一步表征了EEC祖细胞富集的miRNA，miR-7，by in vivodial study以及asex vivoenteroid in mice.ResultsFirst，我们证明了miR-7在整个EEC谱系轨迹中高度富集，并且是EEC祖细胞相对于Lgr 5+肠干细胞中最富集的miRNA。接下来，我们在体内显示，在有利于隐窝分裂和抑制EEC丰度的饮食条件下，EEC祖细胞中的miR-7被显著抑制。然后，我们通过在小鼠类肠中的功能测定证明，miR-7发挥了对生长的鲁棒控制，如通过出芽（隐窝分裂的代理）、EdU和PH 3染色所确定的，并且还可能调节EEC丰度。最后，我们通过单细胞RNA测序分析表明，miR-7 regulatesXiapin祖细胞/干细胞，我们证明了在肠上皮细胞中，miR-7对小鼠肠上皮细胞生长的影响部分依赖于Xip和Egfr signaling.ConclusionsThis研究首次证明，EEC祖细胞富集的miR-7被饮食扰动改变，它通过完整的Xip和Egfr信号调节肠上皮细胞的生长。
Background & AimsThe enteroendocrine cell (EEC) lineage is important for intestinal homeostasis. It was recently shown that EEC progenitors contribute to intestinal epithelial growth and renewal, but the underlying mechanisms remain poorly understood. MicroRNAs are under-explored along the entire EEC lineage trajectory, and comparatively little is known about their contributions to intestinal homeostasis.MethodsWe leverage unbiased sequencing and eight different mouse models and sorting methods to identify microRNAs enriched along the EEC lineage trajectory. We further characterize the functional role of EEC progenitor-enriched miRNA, miR-7, byin vivodietary study as well asex vivoenteroid in mice.ResultsFirst, we demonstrate that miR-7 is highly enriched across the entire EEC lineage trajectory and is the most enriched miRNA in EEC progenitors relative to Lgr5+ intestinal stem cells. Next, we showin vivothat in EEC progenitors miR-7 is dramatically suppressed under dietary conditions that favor crypt division and suppress EEC abundance. We then demonstrate by functional assays in mouse enteroids that miR-7 exerts robust control of growth, as determined by budding (proxy for crypt division), EdU and PH3 staining, and likely regulates EEC abundance also. Finally, we show by single-cell RNA sequencing analysis that miR-7 regulatesXiapin progenitor/stem cells and we demonstrate in enteroids that the effects of miR-7 on mouse enteroid growth depend in part on Xiap and Egfr signaling.ConclusionsThis study demonstrates for the first time that EEC progenitor cell-enriched miR-7 is altered by dietary perturbations and that it regulates growth in enteroids via intact Xiap and Egfr signaling.