MMP Mediated Degradation of Type VI Collagen Is Highly Associated with Liver Fibrosis - Identification and Validation of a Novel Biochemical Marker Assay

Sanne Skovgard Veidal, Morten Asser Karsdal, Efstathios Vassiliadis, Arkadiusz Nawrocki, Martin Rossel Larsen, HTN Quoc, Per Hägglund, Yunyun Luo, Qinlong Zheng, Ben Vainer, Diana Julie Leeming

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    Abstract

    Background and Aims: During fibrogenesis, in which excessive remodeling of the extracellular matrix occurs, both the quantity of type VI collagen and levels of matrix metalloproteinases, including MMP-2 and MMP-9, increase significantly. Proteolytic degradation of type VI collagen into small fragments, so-called neo-epitopes, may be specific biochemical marker of liver fibrosis. The aim of this study was to develop an ELISA detecting a fragment of type VI collagen generated by MMP-2 and MMP-9, and evaluate this assay in two preclinical models of liver fibrosis. Methods: Mass spectrometric analysis of cleaved type VI collagen revealed a large number of protease-generated neo-epitopes. A fragment unique to type VI collagen generated by MMP-2 and MMP-9 was selected for ELISA development. The CO6-MMP assay was evaluated in two rat models of liver fibrosis: bile duct ligation (BDL) and carbon tetrachloride (CCl4)-treated rats. Results: Intra-and inter-assay variation was 4.1% and 10.1% respectively. CO6-MMP levels were significantly elevated in CCl4-treated rats compared to vehicle-treated rats at weeks 12 (mean 30.9 ng/mL vs. 12.8 ng/mL, p = 0.002); week 16 (mean 34.0 ng/mL vs. 13.7 ng/mL, p = 0.0018); and week 20 (mean 35.3 ng/mL vs. 13.3 ng/mL, p = 0.0033) with a tight correlation between hepatic collagen content and serum levels of CO6-MMP (R-2 = 0.58, p
    Original languageEnglish
    JournalP L o S One
    Volume6
    Issue number9
    Pages (from-to)e24753
    ISSN1932-6203
    DOIs
    Publication statusPublished - 2011

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