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Deletion of FHL2 in fibroblasts attenuates fibroblasts activation and kidney fibrosis via restraining TGF-β1-induced Wnt/β-catenin signaling

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Abstract

Four-and-a-half LIM domains protein 2 (FHL2) has been proposed involving in β-catenin activity. We previously reported that FHL2 mediates TGF-β1-induced tubular epithelial-to-mesenchymal transition through activating Wnt/β-catenin signaling. However, the potential role and mechanism for FHL2 in TGF-β1-induced fibroblast activation and kidney fibrosis remains unknown. Here, we initially observed higher levels of FHL2 expression in fibrotic kidneys from both patients and mice, especially in α-smooth muscle actin (α-SMA)-positive cells in the interstitium. In cultured interstitial fibroblasts, FHL2 expression was induced by TGF-β1. Knockdown of FHL2 remarkably suppressed TGF-β1-induced α-SMA, type I collagen, and fibronectin expression, while overexpression of FHL2 was sufficient to activate fibroblasts. In mice, fibroblast-specific deletion of FHL2 diminished renal induction of α-SMA, type I collagen, and fibronectin and interstitial extracellular matrix deposition at 2 weeks after ureteral obstruction. We next investigated Wnt/β-catenin activity and found that β-catenin was activated in most FHL2-positive cells in renal interstitium from mice with obstructive nephropathy. In vitro, TGF-β1 induced a physical interaction between FHL2 and β-catenin, especially in the nucleus. Downregulation of FHL2 inhibited TGF-β1-induced active β-catenin upregulation, β-catenin nuclear translocation, and β-catenin-mediated transcription, whereas overexpression of FHL2 was able to activate Wnt/β-catenin signaling. FHL2 overexpression-induced β-catenin-mediated gene transcription could be hindered by ICG-001, but FHL2 overexpression-induced upregulation of active β-catenin could not be. Collectively, this study reveals that the signal regulatory effect of FHL2 on β-catenin plays an important role in TGF-β1-induced fibroblast activation and kidney fibrosis.

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Author information

HW designed the study and wrote the paper. DY, QY and HX performed the experiments. HW, DY, and QY prepared all figures and analyzed the results. DC and YJ provided valuable suggestions and comments on the study design. All authors approved the final version of the manuscript.

Correspondence to Weichun He.

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The authors declared that they have no conflict of interest.

Ethical approval

Animal protocol was approved by the Institutional Animal Care and Use Committee at the Nanjing Medical University. Study involving human tissues was approved by the Ethics Committee at the Second Affiliated Hospital of Nanjing Medical University.

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Key messages

•FHL2 expression is induced in the interstitial fibroblasts from fibrotic kidneys.

•Fibroblast-specific deletion of FHL2 attenuates interstitial fibrosis in UUO mice.

•FHL2 is responsible for TGF-β1-induced Wnt/β-catenin activity and fibroblasts activation.

•Overexpressing FHL2 is sufficient to activate Wnt/β-catenin signaling and fibroblasts.

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Cite this article

Duan, Y., Qiu, Y., Huang, X. et al. Deletion of FHL2 in fibroblasts attenuates fibroblasts activation and kidney fibrosis via restraining TGF-β1-induced Wnt/β-catenin signaling. J Mol Med (2020) doi:10.1007/s00109-019-01870-1

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Keywords

  • FHL2
  • β-Catenin
  • TGF-β1
  • Fibroblast
  • Kidney fibrosis