Essential roles of YAP-TEAD complex in adult stem cell development during thyroid hormone-induced intestinal remodeling of Xenopus laevis
Essential roles of YAP-TEAD complex in adult stem cell development during thyroid hormone-induced intestinal remodeling of Xenopus laevis
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DOI：
10.1007/s00441-022-03600-5
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发表时间：
2022-02-25
影响因子：
3.6
通讯作者：
Ishizuya-Oka，Atsuko
中科院分区：
文献类型：
--
作者：
Hasebe，Takashi;Fujimoto，Kenta;Ishizuya-Oka，Atsuko
在由甲状腺激素(TH)触发的两栖动物变态过程中，小肠从幼虫广泛重塑为成虫。在非洲爪哇，一些幼虫上皮细胞去分化为成体干细胞，新形成的成体上皮类似于哺乳动物的上皮。我们先前已经证明TH激活的Shh、Wnt和Notch信号通路在成人上皮发育中发挥重要作用。在这里，我们将重点放在河马信号通路上，已知它与哺乳动物肠道中的这些通路相互作用。我们的定量RT-PCR分析表明，在变态肠道中，参与该途径的基因包括YAP1、TAZ、TEAD1和核心激酶的表达受到TH的不同调控。此外，我们通过原位杂交和免疫组织化学方法表明，转录辅助激活因子YAP1是Hippo信号的主要效应因子，在成体干细胞和周围的结缔组织细胞中都有表达，YAP1蛋白定位于干细胞的细胞核或细胞质中。我们进一步证明YAP1在体内与其结合伙伴TEAD1(转录因子)结合，并且它们的相互作用被维替普芬(VP)抑制。更重要的是，通过在蝌蚪肠器官培养中使用VP，我们实验证明，抑制YAP1-TEAD1相互作用既减少了TH诱导的表达LGR5的干细胞，也减少了附近结缔组织细胞的数量和增殖，导致成年上皮发育失败。我们的结果表明，YAP-tead复合体在肠道重塑过程中是干细胞发育所必需的。
During amphibian metamorphosis which is triggered by thyroid hormone (TH), the small intestine is extensively remodeled from the larval to adult form. In theXenopus laevisintestine, some of the larval epithelial cells dedifferentiate into adult stem cells, which newly form the adult epithelium similar to the mammalian one. We have previously shown that TH-activated Shh, Wnt and Notch signaling pathways play important roles in adult epithelial development. Here we focus on the Hippo signaling pathway, which is known to interact with these pathways in the mammalian intestine. Our quantitative RT-PCR analysis indicates that the expression of genes involved in this pathway including YAP1, TAZ, TEAD1 and core kinases is differently regulated by TH in the metamorphosing intestine. Additionally, we show by in situ hybridization and immunohistochemistry that the transcriptional co-activator YAP1, a major effector of the Hippo signaling, is expressed in the adult stem cells and connective tissue cells surrounding them and that YAP1 protein is localized in either nucleus or cytoplasm of the stem cells. We further show that YAP1 binds its binding partner TEAD1 (transcription factor) in vivo and that their interaction is inhibited by verteporfin (VP). More importantly, by using VP in organ culture of the tadpole intestine, we experimentally demonstrate that the inhibition of YAP1-TEAD1 interaction decreases both TH-induced stem cells expressing LGR5 and nearby connective tissue cells in number and proliferation, leading to the failure of adult epithelial development. Our results indicate that YAP-TEAD complex is required for stem cell development during intestinal remodeling.