Inhibition of metalloprotease hyperactivity in cystic cholangiocytes halts the development of polycystic liver diseases

Abstract

Objective Polycystic liver diseases (PCLDs) are genetic disorders characterised by progressive bile duct dilatation and/or cyst development. Their pathogenesis is a consequence of hyperproliferation, hypersecretion and microRNA alterations in cholangiocytes. Here we evaluate the role of matrix metalloproteases (MMPs) in the hepatic cystogenesis of PCLDs. Design Metalloprotease activity was measured by microfluorimetric assays in normal and polycystic cholangiocyte cultures from humans and rats, and gene expression by real time quantitative PCR. The role of cytokines, oestrogens and growth factors present in the cystic fluid of PCLD patients was evaluated for MMP activity. The MMP inhibitor marimastat was examined for cystic expansion in vitro and in polycystic kidney (PCK) rats. Results Polycystic human and rat cholangiocytes displayed increased MMP activity, which was associated with increased mRNA levels of different MMPs. Interleukin (IL)-6 and IL-8, and 17β-oestradiol, all stimulated MMP activity in human cholangiocytes. The presence of antibodies against IL-6 and/or IL-8 receptor/s inhibited baseline MMP hyperactivity of polycystic human cholangiocytes but had no effect on normal human cholangiocytes. MMP-3 was overexpressed in cystic cholangiocytes from PCLD human and PCK rat livers by immunohistochemistry. Marimastat reduced MMP hyperactivity of polycystic human and rat cholangiocytes and blocked the cystic expansion of PCK cholangiocytes cultured in three-dimensions. Chronic treatment of 8-week-old PCK rats with marimastat inhibited hepatic cystogenesis and fibrosis. Conclusions PCLDs are associated with cholangiocyte MMP hyperactivity resulting from autocrine/paracrine stimulation by IL-6 and IL-8. Inhibition of this MMP hyperactivity with marimastat decreased hepatic cystogenesis in vitro and in an animal model of PCLD, offering a potential therapeutic tool.

Integrative Analysis of Transcriptome and MicroRNome Profiles in Cholangiocarcinoma

“…MMP7 has been suggested as an useful biomarker in lung cancer [101] and CCA [98]. MMPs may be targeted by synthetic inhibitors such as Marimastat, a broad spectrum MMP inhibitor [95] that was experimentally tested and shown useful for treatment of polycystic liver disease, which presents cholangiocyte overgrowth by MMP hyperactivity [102]. In addition, Batimastat and Prinomastat are synthetic MMP inhibitors and Neovastat is a natural MMP7 inhibitor [95].…”

Post-translational Regulation of the Type III Inositol 1,4,5-Trisphosphate Receptor by miRNA-506

“…H69 cholangiocytes stably overexpressing miR-506 exhibit fibrotic gene signatures. A, MMP enzymatic activity was assayed in untransfected, control-miR, and miR-506 stably overexpressing H69 human cholangiocyte cells as previously described (n ϭ 12) (24). Quantitative PCR analysis of expression of (B) ␣SMA and (C) CTGF in untransfected, control-miR and miR-506 stably expressing H69 cholangiocyte cells using the primers shown in Table 2 (n ϭ 11-12 per gene).…”

“…Matrix Metalloproteinase activity was measured in wild-type H69, control miR, and miR-506 cells as described by us (24).…”

A novel twist in polycystic liver disease

“…The discovery of an animal model (the PCK rat) that recapitulates many aspects of PLD has fuelled the field, and this signals another key development. 5 Urribarri et al have isolated normal and cyst-derived cholangiocytes and were able to keep these cells over a number of passages while retaining the original features of the cell. They went on to elucidate the role of matrix metalloproteinases (MMPs) in hepatic cystogenesis.…”