PMID- 36085540 OWN - NLM STAT- MEDLINE DCOM- 20220929 LR - 20220929 IS - 1749-0774 (Electronic) IS - 0914-7470 (Linking) VI - 35 IP - 6 DP - 2022 Nov TI - Overexpression of PYGO1 promotes early cardiac lineage development in human umbilical cord mesenchymal stromal/stem cells by activating the Wnt/beta-catenin pathway. PG - 1722-1735 LID - 10.1007/s13577-022-00777-3 [doi] AB - Cardiovascular disease still has the highest mortality. Gene-modified mesenchymal stromal/stem cells could be a promising therapy. Pygo plays an important role in embryonic development and regulates life activities with a variety of regulatory mechanisms. Therefore, this study aimed to investigate whether the overexpression of the PYGO1 gene can promote the differentiation of human umbilical cord-derived mesenchymal stromal/stem cells (HUC-MSCs) into early cardiac lineage cells and to preliminary explore the relevant mechanisms. In this study, HUC-MSCs were isolated by the explant method and were identified by flow cytometry and differentiation assay, followed by transfected with lentivirus carrying the PYGO1 plasmid. In PYGO1 group (cells were incubated with lentiviral-PYGO1), the mRNA expressions of cardiac differentiation-specific markers (MESP1, NKX2.5, GATA4, MEF2C, ISL1, TBX5, TNNT2, ACTC1, and MYH6 genes) and the protein expressions of NKX2.5 and cTnT were significantly up-regulated compared with the NC group (cells were incubated with lentiviral-empty vector). In addition, the proportion of NKX2.5, GATA4, and cTnT immunofluorescence-positive cells increased with the inducement time. Overexpression of PYGO1 statistically significantly increased the relative luciferase expression level of Topflash plasmid, the protein expression level of beta-catenin and the mRNA expression level of CYCLIND1. Compared with the control group, decreased protein levels of NKX2.5 and cTnT were detected in PYGO1 group after application of XAV-939, the specific inhibitor of the canonical Wnt/beta-catenin pathway. Our study suggests that overexpression of PYGO1 significantly promotes the differentiation of HUC-MSCs into early cardiac lineage cells, which is regulated by the canonical Wnt/beta-catenin signaling. CI - (c) 2022. The Author(s) under exclusive licence to Japan Human Cell Society. FAU - Shen, Jie AU - Shen J AD - Department of Pediatrics, The Third Xiangya Hospital, Central South University, Changsha, 410013, Hunan, China. FAU - Wu, Xiushan AU - Wu X AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, China. FAU - Zhu, Ping AU - Zhu P AD - Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, 510080, Guangdong, China. FAU - Zhuang, Jian AU - Zhuang J AD - Guangdong Cardiovascular Institute, Guangdong Provincial People's Hospital, Guangdong Academy of Medical Sciences, Guangzhou, 510080, Guangdong, China. FAU - Qin, Bin AU - Qin B AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, China. FAU - Sun, Fang AU - Sun F AD - Department of Pediatrics, The Third Xiangya Hospital, Central South University, Changsha, 410013, Hunan, China. FAU - Yuan, Wuzhou AU - Yuan W AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, China. FAU - Fan, Xiongwei AU - Fan X AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, China. FAU - Jiang, Zhigang AU - Jiang Z AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, China. FAU - Li, Fang AU - Li F AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, China. FAU - Li, Yongqing AU - Li Y AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, China. FAU - Wang, Yuequn AU - Wang Y AD - The Center for Heart Development, State Key Laboratory of Development Biology of Freshwater Fish, Key Laboratory of MOE for Development Biology and Protein Chemistry, College of Life Sciences, Hunan Normal University, Changsha, 410081, Hunan, China. FAU - Zhao, Mingyi AU - Zhao M AUID- ORCID: 0000-0002-2884-0736 AD - Department of Pediatrics, The Third Xiangya Hospital, Central South University, Changsha, 410013, Hunan, China. 36163773@qq.com. LA - eng GR - 805212639211/Key Program of Guangzhou Science Research/ GR - 81970248/National Natural Science Foundation of China/ GR - 2020zzts888/Fundamental Research Funds for Central Universities of the Central South University/ GR - 2017A030312007/Natural Science Foundation of Guangdong Province/ PT - Journal Article DEP - 20220909 PL - Japan TA - Hum Cell JT - Human cell JID - 8912329 RN - 0 (RNA, Messenger) RN - 0 (beta Catenin) SB - IM MH - Cell Differentiation/genetics MH - Humans MH - *Mesenchymal Stem Cells MH - RNA, Messenger/metabolism MH - Umbilical Cord MH - Wnt Signaling Pathway/genetics MH - *beta Catenin/genetics/metabolism OTO - NOTNLM OT - Cardiac lineage development OT - Human umbilical cord mesenchymal stromal/stem cells OT - PYGO1 OT - Wnt/beta-catenin signaling EDAT- 2022/09/11 06:00 MHDA- 2022/09/30 06:00 CRDT- 2022/09/10 00:07 PHST- 2022/02/20 00:00 [received] PHST- 2022/08/26 00:00 [accepted] PHST- 2022/09/11 06:00 [pubmed] PHST- 2022/09/30 06:00 [medline] PHST- 2022/09/10 00:07 [entrez] AID - 10.1007/s13577-022-00777-3 [pii] AID - 10.1007/s13577-022-00777-3 [doi] PST - ppublish SO - Hum Cell. 2022 Nov;35(6):1722-1735. doi: 10.1007/s13577-022-00777-3. Epub 2022 Sep 9.