2. Academic Validation
  3. Fibrosis resolution in the mouse liver: Role of Mmp12 and potential role of calpain 1/2

Fibrosis resolution in the mouse liver: Role of Mmp12 and potential role of calpain 1/2

  • Matrix Biol Plus. 2022 Dec 28:17:100127. doi: 10.1016/j.mbplus.2022.100127.
Toshifumi Sato 1,  Kimberly Z Head 2,  Jiang Li 1,  Christine E Dolin 3,  Daniel Wilkey 4 5,  Nolan Skirtich 6,  Katelyn Smith 7,  Dylan D McCreary 6,  Sylvia Liu 7 8,  Juliane I Beier 1 8 9,  Aatur D Singhi 7 8,  Ryan M McEnaney 6,  Michael L Merchant 4 5,  Gavin E Arteel 1 8
Affiliations

Affiliations

  • 1 Department of Medicine, Division of Gastroenterology, Hepatology and Nutrition, United States.
  • 2 Department of Pharmacology and Chemical Biology, University of Pittsburgh, Pittsburgh, PA 15213, United States.
  • 3 Department of Pharmacology and Toxicology, University of Louisville, Louisville, KY 40292, United States.
  • 4 Department of Medicine, Division of Nephrology and Hypertension, University of Louisville, Louisville, KY 40292, United States.
  • 5 University of Louisville Alcohol Research Center, University of Louisville, Louisville, KY 40292, United States.
  • 6 Department of Surgery, University of Pittsburgh, Pittsburgh, PA 15213, United States.
  • 7 Department of Pathology, University of Pittsburgh, Pittsburgh, PA 15213, United States.
  • 8 Pittsburgh Liver Research Center, University of Pittsburgh, Pittsburgh, PA 15213, United States.
  • 9 Department of Environmental and Occupational Health, University of Pittsburgh, Pittsburgh, PA 15213, United States.
PMID: 36632559 DOI: 10.1016/j.mbplus.2022.100127
Abstract

Although most work has focused on resolution of Collagen ECM, fibrosis resolution involves changes to several ECM proteins. The purpose of the current study was twofold: 1) to examine the role of MMP12 and elastin; and 2) to investigate the changes in degraded proteins in plasma (i.e., the "degradome") in a preclinical model of fibrosis resolution. Fibrosis was induced by 4 weeks carbon tetrachloride (CCl4) exposure, and recovery was monitored for an additional 4 weeks. Some mice were treated with daily MMP12 inhibitor (MMP408) during the resolution phase. Liver injury and fibrosis was monitored by clinical chemistry, histology and gene expression. The release of degraded ECM peptides in the plasma was analyzed using by 1D-LC-MS/MS, coupled with PEAKS Studio (v10) peptide identification. Hepatic fibrosis and liver injury rapidly resolved in this mouse model. However, some Collagen fibrils were still present 28d after cessation of CCl4. Despite this persistent Collagen presence, expression of canonical markers of fibrosis were also normalized. The inhibition of MMP12 dramatically delayed fibrosis resolution under these conditions. LC-MS/MS analysis identified that several proteins were being degraded even at late stages of fibrosis resolution. Calpains 1/2 were identified as potential new proteases involved in fibrosis resolution. CONCLUSION. The results of this study indicate that remodeling of the liver during recovery from fibrosis is a complex and highly coordinated process that extends well beyond the degradation of the collagenous scar. These results also indicate that analysis of the plasma degradome may yield new insight into the mechanisms of fibrosis recovery, and by extension, new "theragnostic" targets. Lastly, a novel potential role for calpain activation in the degradation and turnover of proteins was identified.

Keywords

CCl4, carbon tetrachloride; ECM, extracellular matrix; ELISA, enzyme-linked immunosorbent assay; Elastin; Fibrosis; GO, gene ontology; LC-MS/MS, liquid Chromatography with tandem mass spectrometry; MMP, matrix metalloprotease; MMP-12; MPM, multiphoton microscopy; SDS, sodium dodecyl sulfate; rtPCR, reverse-transcriptase polymerase chain reaction.

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