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Pyruvate kinase M2 activation protects against the proliferation and migration of pulmonary artery smooth muscle cells

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Abstract

Pyruvate kinase M2 (PKM2), which is encoded by PKM, is a ubiquitously expressed intracellular protein and is associated with proliferation cell phenotype. In PAH patients and PAH models, we found higher levels of PKM2 tyrosine 105 phosphorylation (phospho-PKM2 (Y105)) than in controls, both in vivo and in vitro. Here, we demonstrate that PKM2 stimulates inflammatory and apoptosis signalling pathways in pulmonary artery smooth muscle cells (PASMCs) and promotes PASMC migration and proliferation. PKM2 phosphorylation promoted the dimerization activation and nuclear translocation of STAT3, a transcription factor regulating proliferation, growth, and apoptosis. TLR2, a transmembrane protein receptor involved in both innate and adaptive immune responses, promoted PKM2 phosphorylation in hypoxia-induced PASMCs. Therefore, we hypothesized that PKM2 also affects the proliferation and migration of PASMCs. The proliferation of hypoxia-induced normal human pulmonary artery smooth muscle cells (normal-HPASMCs) was found to be inhibited by TEPP-46 (PKM2 agonist) and PKM2 siRNA using wound healing, 5-ethynyl-2′-deoxyuridine (EdU), and immunofluorescence (Ki67) assays. PASMCs isolated from PAH patients (PAH-HPASMCs) and hypoxia-treated rats (PAH-RPASMCs) also confirmed the above results. TEPP-46 treatment was found to improve hypoxia-induced pulmonary artery remodelling and right heart function in mice, and the link between PKM2 and STAT3 was also confirmed in vivo. In conclusion, PKM2 plays crucial roles in the proliferation and migration of PASMCs.

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Funding

We acknowledge the National Natural Science Foundation of China (NSFC-91639303), Nanjing Municipal Health Bureau (CN)(QRX17016), Department of Human Resources and Social Security of Jiangsu Province (2015-WSN-069), and Natural Science Funds of Jiangsu Province (BK20161112) for supporting completion of this study.

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  1. Aikai Zhang and Fenfang Yu contributed equally to this work.

Authors and Affiliations

  1. Department of Cardiology, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210029, China

    Aikai Zhang, Wande Yu, Peng Ye, Pengfei Liu, Yue Gu, Shaoliang Chen & Hang Zhang

  2. Department of Respiratory, Nanjing First Hospital, Nanjing Medical University, Nanjing, 210029, China

    Fenfang Yu

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  1. Aikai Zhang
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Contributions

ZAK performed the vitro experiment, organized pictures, analysed the data, and drafted the manuscript. YFF and YP performed the animal work, including feeding and harvest. LPF aided in collecting tissues from animals and human. YWD and GY analysed the data and edited the manuscript. ZH and CSL designed and supervised the research and edited the manuscript. The final draft was read and approved by all authors.

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Correspondence to Hang Zhang.

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The authors declare that they have no conflicts of interest.

Ethical approval

All animal experiments were approved by the Institutional Animal Care and Use Committee of Nanjing Medical University. The present study was first approved by the Ethics Committees of the Nanjing Medical University for experiments involving human tissues, and informed consent was obtained from each individual before collecting tissues.

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Zhang, A., Yu, F., Yu, W. et al. Pyruvate kinase M2 activation protects against the proliferation and migration of pulmonary artery smooth muscle cells. Cell Tissue Res 382, 585–598 (2020). https://doi.org/10.1007/s00441-020-03245-2

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