L-glutamate requires β-catenin signalling through Frizzled7 to stimulate porcine intestinal stem cell expansion
L-glutamate requires β-catenin signalling through Frizzled7 to stimulate porcine intestinal stem cell expansion
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DOI：
10.1007/s00018-022-04545-2
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发表时间：
2022-09
影响因子：
8
通讯作者：
Ying-chao Qin;Jia-yi Zhou;Min Zhu;Ge Zan;Chun-qi Gao;Hui-chao Yan;Xiang-guang Li;Xiu-qi Wang
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作者：
Ying-chao Qin;Jia-yi Zhou;Min Zhu;Ge Zan;Chun-qi Gao;Hui-chao Yan;Xiang-guang Li;Xiu-qi Wang
肠道干细胞(ISCs)解码和协调来自饮食的各种营养信息，以支持隐窝-绒毛轴结构，但特定的饮食分子如何影响肠上皮细胞动态平衡仍不清楚。在本研究中，在无氮日粮和玉米-豆粕日粮中添加L-谷氨酸(Glu)，均可刺激断奶仔猪的肠道生长和ISC扩张。定量蛋白质组学筛选发现，典型的Wnt信号通路是体内肠上皮发育和ISC活动的中心调节因子。重要的是，Wnt跨膜受体Frizzled7(FZD7)上调是对饮食中谷氨酸模式的反应，它在用特定抑制剂处理的肠器官(IOS)和FZD7-KOIPEC-J2细胞中的扰动中断了Glu输入和β-catenin信号之间的联系，并随后降低了细胞活力。此外，共定位、免疫共沉淀(Co-IP)、等温滴定量热法(ITC)和微量热电泳法(MST)表明，Glu是直接与FZD7结合的信号分子。我们认为，FZD7介导的细胞外Glu信号的整合控制着ISC的增殖和分化，这为研究营养物质与ISCs的相互作用提供了新的见解。
Intestinal stem cells (ISCs) decode and coordinate various types of nutritional information from the diet to support the crypt–villus axis architecture, but how specific dietary molecules affect intestinal epithelial homeostasis remains unclear. In the current study, L-glutamate (Glu) supplementation in either a nitrogen-free diet (NFD) or a corn-soybean meal diet (CSMD) stimulated gut growth and ISC expansion in weaned piglets. Quantitative proteomics screening identified the canonical Wnt signalling pathway as a central regulator of intestinal epithelial development and ISC activity in vivo. Importantly, the Wnt transmembrane receptor Frizzled7 (FZD7) was upregulated in response to dietary Glu patterns, and its perturbations in intestinal organoids (IOs) treated with a specific inhibitor and in FZD7-KO IPEC-J2 cells disrupted the link between Glu inputs and β-catenin signalling and a subsequent reduction in cell viability. Furthermore, co-localization, coimmunoprecipitation (Co-IP), isothermal titration calorimetry (ITC), and microscale thermophoresis (MST) revealed that Glu served as a signalling molecule directly bound to FZD7. We propose that FZD7-mediated integration of the extracellular Glu signal controls ISC proliferation and differentiation, which provides new insights into the crosstalk of nutrients and ISCs.