Mmp14-controlled extracellular matrix homeostasis is required for circadian rhythm

GARVA, R.; Yeung, C.-Y. C.; Pickard, A.; Lu, Y.; Mallikarjun, V.; Swift, J.; Taylor, S. H.; Rai, J.; Eyre, D. R.; Chaturvedi, M.; Itoh, Y.; Meng, Q.-J.; Mauch, C.; Zigrino, P.; Kadler, K. E.

doi:10.1101/2022.04.01.486699

bioRxiv · 10.1101/2022.04.01.486699

Mmp14-controlled extracellular matrix homeostasis is required for circadian rhythm

GARVA, R.·Yeung, C.-Y. C.·Pickard, A.·Lu, Y.·Mallikarjun, V.·Swift, J.·Taylor, S. H.·Rai, J.·Eyre, D. R.·Chaturvedi, M.·Itoh, Y.·Meng, Q.-J.·Mauch, C.·Zigrino, P.·Kadler, K. E.

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Abstract

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.

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GARVA, R., Yeung, C.-Y. C., Pickard, A., Lu, Y., Mallikarjun, V., Swift, J., Taylor, S. H., Rai, J., Eyre, D. R., Chaturvedi, M., Itoh, Y., Meng, Q.-J., Mauch, C., Zigrino, P., Kadler, K. E.. 2022-04-02. Mmp14-controlled extracellular matrix homeostasis is required for circadian rhythm. https://doi.org/10.1101/2022.04.01.486699

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