Xiao Yao, Wang Yujie, Zhang Minghao, Zhang Yan, Ju Zhihua, Wang Jinpeng, Zhang Yaran, Yang Chunhong, Wang Xiuge, Jiang Qiang, Gao Yaping, Wei Xiaochao, Liu Wenhao, Gao Yundong, Hu Peng, Huang Jinming
Key Laboratory of Livestock and Poultry Multi-omics of Ministry of Agriculture and Rural Affairs, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China; Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China; Technical Innovation Center of Dairy Cattle Breeding Industry of Shandong Province, Jinan, 250100, China.
Key Laboratory of Livestock and Poultry Multi-omics of Ministry of Agriculture and Rural Affairs, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China; Shandong Key Laboratory of Animal Disease Control and Breeding, Institute of Animal Science and Veterinary Medicine, Shandong Academy of Agricultural Sciences, Jinan, 250100, China; College of Life Sciences, Shandong Normal University, Jinan, 250358, China.
Theriogenology. 2025 Feb;233:100-111. doi: 10.1016/j.theriogenology.2024.11.010. Epub 2024 Nov 19.
The use of tankyrase inhibitors is essential for capturing livestock embryonic stem cells (ESC), yet their mechanisms of action remain largely uncharacterized. Previous studies indicate that their roles extend beyond the suppression of canonical WNT signaling. This study investigates the effects of the tankyrase inhibitor IWR-1 on maintaining the pluripotency of bovine embryonic stem cells (bESC) cultured on mitotically inactivated mouse embryonic fibroblasts (MEF). Notably, bESC exhibited significant differentiation after one month of IWR-1 withdrawal, without a clear bias toward any specific germ layer. IWR-1 effectively inhibited TNKS2 activity in bESC, whereas it suppressed TNKS1 protein level in growth-arrested MEF. Early differentiation upon IWR-1 removal induced more substantial transcriptomic changes in MEF than in bESC. Furthermore, cell communication analysis predicted that IWR-1 influenced several paracrine and autocrine signals within the culture system. We also observed that IWR-1 repressed protein abundance of the HIPPO pathway components including TEAD4 and YAP1 in bESC and decreased transcription of HIPPO targeted genes CYR61. Protein analysis in growth-arrested MEF suggested that IWR-1 modulated MEF function by impeding TGF-β1 activation and activin A secretion which mitigated nuclear localization of SMAD2/3 in the bESC. This study underscores the role of tankyrase inhibitors in ESC self-renewal by modulating key signaling pathways and orchestrating cell-cell interactions, which may be meaningful in understanding the delicate signaling control of pluripotency in livestock and improving the culture system.
端锚聚合酶抑制剂的使用对于捕获家畜胚胎干细胞（ESC）至关重要，但其作用机制在很大程度上仍未明确。先前的研究表明，它们的作用超出了对经典WNT信号通路的抑制。本研究调查了端锚聚合酶抑制剂IWR-1对维持在有丝分裂失活的小鼠胚胎成纤维细胞（MEF）上培养的牛胚胎干细胞（bESC）多能性的影响。值得注意的是，在撤去IWR-1一个月后，bESC表现出明显的分化，且没有明显偏向任何特定胚层。IWR-1有效抑制了bESC中的TNKS2活性，而在生长停滞的MEF中它抑制了TNKS1蛋白水平。去除IWR-1后的早期分化在MEF中诱导的转录组变化比在bESC中更显著。此外，细胞通讯分析预测IWR-1影响了培养系统内的几种旁分泌和自分泌信号。我们还观察到IWR-1抑制了bESC中包括TEAD4和YAP1在内的HIPPO信号通路成分的蛋白丰度，并降低了HIPPO靶向基因CYR61的转录。对生长停滞的MEF进行的蛋白分析表明，IWR-1通过阻碍TGF-β1激活和激活素A分泌来调节MEF功能，从而减轻了bESC中SMAD2/3的核定位。本研究强调了端锚聚合酶抑制剂通过调节关键信号通路和协调细胞间相互作用在ESC自我更新中的作用，这对于理解家畜多能性的精细信号控制和改进培养系统可能具有重要意义。
