PMID- 37233752 OWN - NLM STAT- MEDLINE DCOM- 20230703 LR - 20230704 IS - 1432-0878 (Electronic) IS - 0302-766X (Print) IS - 0302-766X (Linking) VI - 393 IP - 1 DP - 2023 Jul TI - Novel biphasic mechanism of the canonical Wnt signalling component PYGO2 promotes cardiomyocyte differentiation from hUC-MSCs. PG - 163-179 LID - 10.1007/s00441-023-03774-6 [doi] AB - Human umbilical cord-derived mesenchymal stem cells (hUC-MSCs) are used to regenerate the myocardium during cardiac repair after myocardial infarction. However, the regulatory mechanism underlying their ability to form mesodermal cells and differentiate into cardiomyocytes remains unclear. Here, we established a human-derived MSCs line isolated from healthy umbilical cords and established a cell model of the natural state to examine the differentiation of hUC-MSCs into cardiomyocytes. Quantitative RT-PCR, western blotting, immunofluorescence, flow cytometry, RNA Seq, and inhibitors of canonical Wnt signalling were used to detect the germ-layer markers T and MIXL1; the markers of cardiac progenitor cells MESP1, GATA4, and NKX2.5 and the cardiomyocyte-marker cTnT to identify the molecular mechanism associated with PYGO2, a key component of the canonical Wnt signalling pathway that regulates the formation of cardiomyocyte-like cells. We demonstrated that PYGO2 promotes the formation of mesodermal-like cells and their differentiation into cardiomyocytes through the hUC-MSC-dependent canonical Wnt signalling by promoting the early-stage entry of beta-catenin into the nucleus. Surprisingly, PYGO2 did not alter the expression of the canonical-Wnt, NOTCH, or BMP signalling pathways during the middle-late stages. In contrast, PI3K-Akt signalling promoted hUC-MSCs formation and their differentiation into cardiomyocyte-like cells. To the best of our knowledge, this is the first study to demonstrate that PYGO2 uses a biphasic mechanism to promote cardiomyocyte formation from hUC-MSCs. CI - (c) 2023. The Author(s). FAU - Shi, Yan AU - Shi Y AD - Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. AD - Laboratory of Artificial Intelligence and 3D Technologies for Cardiovascular Diseases, Guangdong Provincial Key Laboratory of South China Structural Heart Disease, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, 510080, People's Republic of China. FAU - Qin, Bin AU - Qin B AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Hunan, Changsha, 410081, China. FAU - Fan, Xiongwei AU - Fan X AD - Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. FAU - Li, Yongqing AU - Li Y AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Hunan, Changsha, 410081, China. FAU - Wang, Yuequn AU - Wang Y AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Hunan, Changsha, 410081, China. FAU - Yuan, Wuzhou AU - Yuan W AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Hunan, Changsha, 410081, China. FAU - Jiang, Zhigang AU - Jiang Z AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Hunan, Changsha, 410081, China. FAU - Zhu, Ping AU - Zhu P AD - Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. AD - Guangdong Provincial Key Laboratory of Pathogenesis, Targeted Prevention and Treatment of Heart Disease, Guangzhou, Guangdong, 510080, People's Republic of China. FAU - Chen, Jimei AU - Chen J AD - Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. FAU - Chen, Yu AU - Chen Y AD - Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. AD - Guangdong Provincial Key Laboratory of Pathogenesis, Targeted Prevention and Treatment of Heart Disease, Guangzhou, Guangdong, 510080, People's Republic of China. FAU - Li, Fang AU - Li F AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Hunan, Changsha, 410081, China. FAU - Wan, Yongqi AU - Wan Y AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Hunan, Changsha, 410081, China. 954906876@qq.com. FAU - Wu, Xiushan AU - Wu X AUID- ORCID: 0000-0001-8431-4266 AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, College of Life Sciences, Hunan Normal University, Hunan, Changsha, 410081, China. xiushanwu2003@aliyun.com. AD - Guangdong Provincial Key Laboratory of Pathogenesis, Targeted Prevention and Treatment of Heart Disease, Guangzhou, Guangdong, 510080, People's Republic of China. xiushanwu2003@aliyun.com. FAU - Zhuang, Jian AU - Zhuang J AD - Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, Guangdong 510100, China. zhuangjiangenetics@163.com. AD - Laboratory of Artificial Intelligence and 3D Technologies for Cardiovascular Diseases, Guangdong Provincial Key Laboratory of South China Structural Heart Disease, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, 510080, People's Republic of China. zhuangjiangenetics@163.com. LA - eng GR - 82100321/National Natural Science Foundation of China/ GR - 81670290/National Natural Science Foundation of China/ GR - 81970324/National Natural Science Foundation of China/ GR - 81974019/National Natural Science Foundation of China/ GR - 2020M682642/Postdoctoral Research Foundation of China/ GR - 2019B020230003/Science and Technology Planning Project of Guangdong Province/ GR - 2018B090944002/Science and Technology Planning Project of Guangdong Province/ GR - 2020B1111170011/Science and Technology Planning Project of Guangdong Province/ GR - 2022B1212010010/Science and Technology Planning Project of Guangdong Province/ GR - 201904020047/Key Program of Guangzhou Science Research Plan/ GR - 202206010049/Technology Program of Guangzhou/ GR - DFJH201802/Guangdong Peak Project/ GR - 2018YFA0108700/National Key Research and Development Program of China/ GR - 2017YFA0105602/National Key Research and Development Program of China/ GR - 81720108004/NSFC Projects of International Cooperation and Exchanges/ GR - 2017A030312007/Research Team Project of Natural Science Foundation of Guangdong Province of China/ GR - DFJH201812/Special Project of Dengfeng Program of Guangdong Provincial People's Hospital/ GR - KJ012019119/Special Project of Dengfeng Program of Guangdong Provincial People's Hospital/ GR - KJ012019423/Special Project of Dengfeng Program of Guangdong Provincial People's Hospital/ PT - Journal Article DEP - 20230526 PL - Germany TA - Cell Tissue Res JT - Cell and tissue research JID - 0417625 RN - EC 2.7.1.- (Phosphatidylinositol 3-Kinases) RN - 0 (PYGO2 protein, human) RN - 0 (Intracellular Signaling Peptides and Proteins) SB - IM MH - Humans MH - Myocytes, Cardiac/metabolism MH - Phosphatidylinositol 3-Kinases/metabolism MH - *Mesenchymal Stem Cells MH - Wnt Signaling Pathway MH - Cell Differentiation MH - Umbilical Cord MH - *Mesenchymal Stem Cell Transplantation MH - Intracellular Signaling Peptides and Proteins/metabolism PMC - PMC10313541 OTO - NOTNLM OT - Canonical Wnt signalling pathway OT - Cardiomyocyte OT - Human umbilical cord-derived mesenchymal stem cells OT - PI3K-Akt signalling OT - PYGO2 OT - beta-catenin COIS- The authors declare no competing interests. EDAT- 2023/05/26 13:09 MHDA- 2023/07/03 06:41 PMCR- 2023/05/26 CRDT- 2023/05/26 11:03 PHST- 2022/08/30 00:00 [received] PHST- 2023/04/19 00:00 [accepted] PHST- 2023/07/03 06:41 [medline] PHST- 2023/05/26 13:09 [pubmed] PHST- 2023/05/26 11:03 [entrez] PHST- 2023/05/26 00:00 [pmc-release] AID - 10.1007/s00441-023-03774-6 [pii] AID - 3774 [pii] AID - 10.1007/s00441-023-03774-6 [doi] PST - ppublish SO - Cell Tissue Res. 2023 Jul;393(1):163-179. doi: 10.1007/s00441-023-03774-6. Epub 2023 May 26.