Redundant Sources of Wnt Regulate Intestinal Stem Cells and Promote Formation of Paneth Cells
Redundant Sources of Wnt Regulate Intestinal Stem Cells and Promote Formation of Paneth Cells
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DOI：
10.1053/j.gastro.2012.08.031
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发表时间：
2012-12-01
期刊：
影响因子：
29.4
通讯作者：
Clevers, Hans
中科院分区：
文献类型：
--
作者：
Farin, Henner F.;Van Es, Johan H.;Clevers, Hans
背景与目的：Wnt信号调节肠道生理的多个方面，包括干细胞的维持。潘氏细胞通过分泌Wnt支持干细胞，但对单个Wnt家族成员的确切来源和主要功能知之甚少。方法：我们分析了肠组织和培养的上皮细胞从成年小鼠与Wnt 3的条件性缺失（Vil-CreERT 2; Wnt 3fl/fl小鼠）。我们还分析了缺乏分泌细胞的ARAM 1突变小鼠的肠组织和细胞。结果：出乎意料的是，Wnt 3在小鼠肠道干细胞的维持中被抑制，表明Wnt信号的冗余。相比之下，培养的隐窝类器官需要潘氏细胞衍生的Wnt 3。添加外源性Wnt或与间充质细胞共培养恢复了Vil-CreERT 2; Wnt 3fl/fl隐窝类器官的生长。来自ARAM 1突变小鼠的肠类器官不生长或形成潘氏细胞;添加Wnt 3允许在不存在潘氏细胞的情况下生长。Wnt信号传导与Lgr 4/5配体R-spondin具有协同效应以诱导潘氏细胞的形成。嵌合表达的Wnt 3在类器官中使用逆转录病毒载体促进分化的潘氏细胞在细胞自主的方式。结论：Wnt是影响小鼠肠干细胞和潘氏细胞稳态的信号环的一部分。Wnt 3信号是类器官培养物生长和发育所必需的，而非上皮Wnt信号可以提供Wnt的第二生理来源。
BACKGROUND & AIMS: Wnt signaling regulates multiple aspects of intestinal physiology, including stem cell maintenance. Paneth cells support stem cells by secreting Wnt, but little is known about the exact sources and primary functions of individual Wnt family members. METHODS: We analyzed intestinal tissues and cultured epithelial cells from adult mice with conditional deletion of Wnt3(Vil-CreERT2;Wnt3fl/fl mice). We also analyzed intestinal tissues and cells from Atob1 mutant mice, which lack secretory cells. RESULTS: Unexpectedly, Wnt3 was dispensable for maintenance of intestinal stem cells in mice, indicating a redundancy of Wnt signals. By contrast, cultured crypt organoids required Paneth cell-derived Wnt3. Addition of exogenous Wnt, or coculture with mesenchymal cells, restored growth of Vil-CreERT2;Wnt3fl/fl crypt organoids. Intestinal organoids from Atob1 mutant mice did not grow or form Paneth cells; addition of Wnt3 allowed growth in the absence of Paneth cells. Wnt signaling had a synergistic effect with the Lgr4/5 ligand R-spondin to induce formation of Paneth cells. Mosaic expression of Wnt3 in organoids using a retroviral vector promoted differentiation of Paneth cells in a cell-autonomous manner. CONCLUSIONS: Wnt is part of a signaling loop that affects homeostasis of intestinal stem and Paneth cells in mice. Wnt3 signaling is required for growth and development of organoid cultures, whereas nonepithelial Wnt signals could provide a secondary physiological source of Wnt.