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Overexpression of PYGO1 promotes early cardiac lineage development in human umbilical cord mesenchymal stromal/stem cells by activating the Wnt/β-catenin pathway

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Abstract

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 β-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/β-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/β-catenin signaling.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This study was funded by grants of the National Natural Science Foundation of China (81970248); the Graduate research innovation project of Central South University (2020zzts888); the Research Team Project of Natural Science Foundation of Guangdong Province of China (2017A030312007), and the Key Program of Guangzhou Science Research Plan (805212639211).

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Authors and Affiliations

  1. Department of Pediatrics, The Third Xiangya Hospital, Central South University, Changsha, 410013, Hunan, China

    Jie Shen, Fang Sun & Mingyi Zhao

  2. 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

    Xiushan Wu, Bin Qin, Wuzhou Yuan, Xiongwei Fan, Zhigang Jiang, Fang Li, Yongqing Li & Yuequn Wang

  3. Guangdong Cardiovascular Institute, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical Sciences, Guangzhou, 510080, Guangdong, China

    Ping Zhu & Jian Zhuang

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  1. Jie Shen
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Contributions

JS: performed the experiments, analyzed and interpreted the data, and wrote the initial manuscript. XW, PZ, and JZ: conceived the experiments. BQ and FS: provided assistances for experiments. WY, XF, ZJ, FL, YL, and YW: contributed to manuscript correction. MZ: conceptualized and guaranteed the research. All authors approved the final manuscript as submitted and agreed to be accountable for all aspects of the work.

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Correspondence to Mingyi Zhao.

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This study was performed in line with the principles of the Declaration of Helsinki. Approval was granted by the Ethics Committee of the third Xiangya hospital of Central South University (No: 20042).

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Shen, J., Wu, X., Zhu, P. et al. Overexpression of PYGO1 promotes early cardiac lineage development in human umbilical cord mesenchymal stromal/stem cells by activating the Wnt/β-catenin pathway. Human Cell 35, 1722–1735 (2022). https://doi.org/10.1007/s13577-022-00777-3

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