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Wnt4 negatively regulates the TGF-β1-induced human dermal fibroblast-to-myofibroblast transition via targeting Smad3 and ERK

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Abstract

Abnormal activation of Wnt signaling has been demonstrated in the wound healing process and the pathogenesis of fibrotic disorders, with Wnt4 specifically identified as having a key role in the pathogenesis of renal, pulmonary and liver fibrosis. Wnt4 also was found to be upregulated by transforming growth factor-β1 (TGF-β1) in fetal and postnatal murine fibroblasts and bone marrow mesenchymal cells, suggesting an underlying cooperation between Wnt4 and TGF-β1 in fibrosis. However, the specific roles of Wnt4 in TGF-β1-induced skin myofibroblast transition and hypertrophic scar formation remain unclear. In the present study, we first observed reduced Wnt4 expression in hypertrophic scar tissue compared with that in normal skin tissue. Following upregulation by TGF-β1, Wnt4 inhibited the TGF-β1-induced transdifferentiation of fibroblasts into myofibroblasts. Using fibroblast-populated collagen lattice contraction assays, we showed that the increased contractility induced by TGF-β1 was significantly blocked by exogenous Wnt4 and the α-smooth muscle actin (α-SMA) expression was decreased in fibroblasts in the collagen lattices. In addition, knockdown of Wnt4 resulted in further increases in α-SMA and collagen I expressions. Further investigation showed that Wnt4 could inhibit the autocrine effect of TGF-β1 as well as block the phosphorylation of Smad3 and ERK but not of AKT or JNK. Lastly, using hypertrophic scar–derived fibroblasts, we showed that the elevated α-SMA and collagen I levels were markedly reduced after treatment with Wnt4. Taken together, our results suggest that Wnt4 negatively regulates TGF-β1-induced fibroblast activation, which may represent a novel therapeutic strategy for the treatment and prevention of hypertrophic scars.

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Funding

This work was supported by the National Natural Science Foundation of China (No. 81201470, No. 81772071, No. 81501684), the Natural Science Foundation of Shaanxi Province (No. 2017JQ8031) and the China Postdoctoral Science Foundation (No. 2014M562600).

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Author notes

  1. Jiaqi Liu, Bin Zhao, and Huayu Zhu contributed equally to this work.

Authors and Affiliations

  1. Department of Burns and Cutaneous Surgery, Xijing Hospital, The Fourth Military Medical University, 127 Changle West Road, Xi’an, 710032, Shaanxi, China

    Jiaqi Liu, Bin Zhao, Huayu Zhu, Xiaozhi Bai, Xiaoqiang Li, Xiaolong Hu, Min Zhang, Jihong Shi, Zhao Zheng & Dahai Hu

  2. School of Life Science, Northwestern Polytechnical University, Xi’an, 710072, Shaanxi, China

    Bin Zhao

  3. Outpatient Department, The Second Artillery Engineering College, 18 Lishan Road, Xi’an, 710025, Shaanxi, China

    Qing Pan

  4. The PLA 94062 troops Korla Air Force Hospital, Korla, 841000, Xinjiang, China

    Mingda Cai

  5. Department of Immunology, School of Preclinical Education, The Fourth Military Medical University, 127 Changle West Road, Xi’an, 710032, Shaanxi, China

    Angang Yang

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  1. Jiaqi Liu
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Correspondence to Angang Yang or Dahai Hu.

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Written informed consent was obtained from each patient.

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All applicable international, national and/or institutional guidelines for the care and use of animals were followed. All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards. All the experiments, including the collection of skin and scar tissue samples from patients, were approved by the ethics committee of Xijing Hospital, The Fourth Military Medical University (Approval No. KY20183245).

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Liu, J., Zhao, B., Zhu, H. et al. Wnt4 negatively regulates the TGF-β1-induced human dermal fibroblast-to-myofibroblast transition via targeting Smad3 and ERK. Cell Tissue Res 379, 537–548 (2020). https://doi.org/10.1007/s00441-019-03110-x

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