Mmp14 is required for matrisome homeostasis and circadian rhythm in fibroblasts

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  • Yeung, Ching-Yan Chloé
  • Richa Garva
  • Adam Pickard
  • Yinhui Lu
  • Venkatesh Mallikarjun
  • Joe Swift
  • Susan H. Taylor
  • Jyoti Rai
  • David R. Eyre
  • Mayank Chaturvedi
  • Yoshifumi Itoh
  • Qing Jun Meng
  • Cornelia Mauch
  • Paola Zigrino
  • Karl E. Kadler

The circadian clock in tendon regulates the daily rhythmic synthesis of collagen-I and the appearance and disappearance of small-diameter collagen fibrils in the extracellular matrix. How the fibrils are assembled and removed is not fully understood. Here, we first showed that the collagenase, membrane type I-matrix metalloproteinase (MT1-MMP, encoded by Mmp14), is regulated by the circadian clock in postnatal mouse tendon. Next, we generated tamoxifen-induced Col1a2-Cre-ERT2::Mmp14 KO mice (Mmp14 conditional knockout (CKO)). The CKO mice developed hind limb dorsiflexion and thickened tendons, which accumulated narrow-diameter collagen fibrils causing ultrastructural disorganization. Mass spectrometry of control tendons identified 1195 proteins of which 212 showed time-dependent abundance. In Mmp14 CKO mice 19 proteins had reversed temporal abundance and 176 proteins lost time dependency. Among these, the collagen crosslinking enzymes lysyl oxidase-like 1 (LOXL1) and lysyl hydroxylase 1 (LH1; encoded by Plod2) were elevated and had lost time-dependent regulation. High-pressure chromatography confirmed elevated levels of hydroxylysine aldehyde (pyridinoline) crosslinking of collagen in CKO tendons. As a result, collagen-I was refractory to extraction. We also showed that CRISPR-Cas9 deletion of Mmp14 from cultured fibroblasts resulted in loss of circadian clock rhythmicity of period 2 (PER2), and recombinant MT1-MMP was highly effective at cleaving soluble collagen-I but less effective at cleaving collagen pre-assembled into fibrils. In conclusion, our study shows that circadian clock-regulated Mmp14 controls the rhythmic synthesis of small diameter collagen fibrils, regulates collagen crosslinking, and its absence disrupts the circadian clock and matrisome in tendon fibroblasts.

Original languageEnglish
JournalMatrix Biology
Pages (from-to)8-22
Number of pages15
Publication statusPublished - 2023

Bibliographical note

Publisher Copyright:
© 2023 The Author(s)

    Research areas

  • Cell surface enzyme, Circadian rhythm, Collagen, Connective tissue, CRISPR/Cas-9, Electron microscopy, Gene knockout, Matrix metalloproteinase (MMP), Protein self-assembly, Tendon

ID: 374665447