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Hepatocyte produced matrix metalloproteinases are regulated by CD147 in liver fibrogenesis.

Calabro SR, Maczurek AE, Morgan AJ, Tu T, Wen VW, Yee C, Mridha A, Lee M, d'Avigdor W, Locarnini SA, McCaughan GW, Warner FJ, McLennan SV, Shackel NA - PLoS ONE (2014)

Bottom Line: Functionally significant in-vitro regulation of hepatocyte MMP production by CD147 was demonstrated using siRNA to CD147 that decreased hepatocyte MMP-2 and -9 expression/activity.Targeting CD147 regulates hepatocyte MMP production both in-vitro and in-vivo, with the net result being reduced fibrotic matrix turnover in-vivo.Therefore, CD147 regulation of hepatocyte MMP is a novel pathway that could be targeted by future anti-fibrogenic agents.

View Article: PubMed Central - PubMed

Affiliation: Liver Cell Biology, Centenary Institute, Sydney, NSW, Australia; Sydney Medical School, The University of Sydney, Sydney, NSW, Australia.

ABSTRACT

Background: The classical paradigm of liver injury asserts that hepatic stellate cells (HSC) produce, remodel and turnover the abnormal extracellular matrix (ECM) of fibrosis via matrix metalloproteinases (MMPs). In extrahepatic tissues MMP production is regulated by a number of mechanisms including expression of the glycoprotein CD147. Previously, we have shown that CD147 is expressed on hepatocytes but not within the fibrotic septa in cirrhosis [1]. Therefore, we investigated if hepatocytes produce MMPs, regulated by CD147, which are capable of remodelling fibrotic ECM independent of the HSC.

Methods: Non-diseased, fibrotic and cirrhotic livers were examined for MMP activity and markers of fibrosis in humans and mice. CD147 expression and MMP activity were co-localised by in-situ zymography. The role of CD147 was studied in-vitro with siRNA to CD147 in hepatocytes and in-vivo in mice with CCl4 induced liver injury using ãCD147 antibody intervention.

Results: In liver fibrosis in both human and mouse tissue MMP expression and activity (MMP-2, -9, -13 and -14) increased with progressive injury and localised to hepatocytes. Additionally, as expected, MMPs were abundantly expressed by activated HSC. Further, with progressive fibrosis there was expression of CD147, which localised to hepatocytes but not to HSC. Functionally significant in-vitro regulation of hepatocyte MMP production by CD147 was demonstrated using siRNA to CD147 that decreased hepatocyte MMP-2 and -9 expression/activity. Further, in-vivo α-CD147 antibody intervention decreased liver MMP-2, -9, -13, -14, TGF-β and α-SMA expression in CCl4 treated mice compared to controls.

Conclusion: We have shown that hepatocytes produce active MMPs and that the glycoprotein CD147 regulates hepatocyte MMP expression. Targeting CD147 regulates hepatocyte MMP production both in-vitro and in-vivo, with the net result being reduced fibrotic matrix turnover in-vivo. Therefore, CD147 regulation of hepatocyte MMP is a novel pathway that could be targeted by future anti-fibrogenic agents.

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Expression of CD147 mRNA in human liver.Quantitative PCR on whole liver tissue, non-diseased as well as PBC, AIH and HCV cirrhosis of CD147 (splice variant 2) mRNA (n = 4 per group). CD147 was significantly increased in all cirrhotic specimens compared to non-diseased controls, *p<0.05.
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pone-0090571-g004: Expression of CD147 mRNA in human liver.Quantitative PCR on whole liver tissue, non-diseased as well as PBC, AIH and HCV cirrhosis of CD147 (splice variant 2) mRNA (n = 4 per group). CD147 was significantly increased in all cirrhotic specimens compared to non-diseased controls, *p<0.05.

Mentions: CD147 is a known regulator of MMPs, enzymes that play an important role in the remodelling of the ECM during liver injury and cirrhosis [3], [6], [41], [42]. We therefore wished to investigate whether MMPs within the injured liver would also be regulated by CD147. To quantitatively compare CD147 expression between non-diseased and cirrhotic liver tissues we used qPCR. A significant increase of greater than 2 fold in CD147 mRNA levels was seen in PBC, AIH and HCV explanted liver tissues compared to non-diseased liver tissue (Figure 4, p<0.05 and n = 4 per group). To further characterize CD147 localisation within the liver we stained non-diseased liver and end-stage cirrhotic liver with α-CD147 antibody. Figure 5 Panels A and D show CD147 expression in non-diseased tissue, which was equally distributed across the liver, while CD147 within the cirrhotic liver localised to cirrhotic nodules and not the fibrous bands (Figure 5 Panels B and E). No staining was observed in the isotype controls (Figure 5 Panels C and F). Figure 5 shows non-diseased donor and cirrhotic tissue from an ALD explant, we found identical staining for further non-diseased samples as well as PBC, PSC, HCV and AIH tissues (n = 3 per group, not shown). This staining pattern was also observed in murine models of liver injury (CCl4 and thioacetamide) using the commercially available and well characterized α-CD147 antibodies RL73.2 or G19 (data not shown). In non-diseased and cirrhotic liver, CD147 immuno-reactivity was consistent with cell membrane expression in hepatocytes. Importantly, no CD147 immuno-reactivity was observed in HSC within the fibrotic septa.


Hepatocyte produced matrix metalloproteinases are regulated by CD147 in liver fibrogenesis.

Calabro SR, Maczurek AE, Morgan AJ, Tu T, Wen VW, Yee C, Mridha A, Lee M, d'Avigdor W, Locarnini SA, McCaughan GW, Warner FJ, McLennan SV, Shackel NA - PLoS ONE (2014)

Expression of CD147 mRNA in human liver.Quantitative PCR on whole liver tissue, non-diseased as well as PBC, AIH and HCV cirrhosis of CD147 (splice variant 2) mRNA (n = 4 per group). CD147 was significantly increased in all cirrhotic specimens compared to non-diseased controls, *p<0.05.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4116334&req=5

pone-0090571-g004: Expression of CD147 mRNA in human liver.Quantitative PCR on whole liver tissue, non-diseased as well as PBC, AIH and HCV cirrhosis of CD147 (splice variant 2) mRNA (n = 4 per group). CD147 was significantly increased in all cirrhotic specimens compared to non-diseased controls, *p<0.05.
Mentions: CD147 is a known regulator of MMPs, enzymes that play an important role in the remodelling of the ECM during liver injury and cirrhosis [3], [6], [41], [42]. We therefore wished to investigate whether MMPs within the injured liver would also be regulated by CD147. To quantitatively compare CD147 expression between non-diseased and cirrhotic liver tissues we used qPCR. A significant increase of greater than 2 fold in CD147 mRNA levels was seen in PBC, AIH and HCV explanted liver tissues compared to non-diseased liver tissue (Figure 4, p<0.05 and n = 4 per group). To further characterize CD147 localisation within the liver we stained non-diseased liver and end-stage cirrhotic liver with α-CD147 antibody. Figure 5 Panels A and D show CD147 expression in non-diseased tissue, which was equally distributed across the liver, while CD147 within the cirrhotic liver localised to cirrhotic nodules and not the fibrous bands (Figure 5 Panels B and E). No staining was observed in the isotype controls (Figure 5 Panels C and F). Figure 5 shows non-diseased donor and cirrhotic tissue from an ALD explant, we found identical staining for further non-diseased samples as well as PBC, PSC, HCV and AIH tissues (n = 3 per group, not shown). This staining pattern was also observed in murine models of liver injury (CCl4 and thioacetamide) using the commercially available and well characterized α-CD147 antibodies RL73.2 or G19 (data not shown). In non-diseased and cirrhotic liver, CD147 immuno-reactivity was consistent with cell membrane expression in hepatocytes. Importantly, no CD147 immuno-reactivity was observed in HSC within the fibrotic septa.

Bottom Line: Functionally significant in-vitro regulation of hepatocyte MMP production by CD147 was demonstrated using siRNA to CD147 that decreased hepatocyte MMP-2 and -9 expression/activity.Targeting CD147 regulates hepatocyte MMP production both in-vitro and in-vivo, with the net result being reduced fibrotic matrix turnover in-vivo.Therefore, CD147 regulation of hepatocyte MMP is a novel pathway that could be targeted by future anti-fibrogenic agents.

View Article: PubMed Central - PubMed

Affiliation: Liver Cell Biology, Centenary Institute, Sydney, NSW, Australia; Sydney Medical School, The University of Sydney, Sydney, NSW, Australia.

ABSTRACT

Background: The classical paradigm of liver injury asserts that hepatic stellate cells (HSC) produce, remodel and turnover the abnormal extracellular matrix (ECM) of fibrosis via matrix metalloproteinases (MMPs). In extrahepatic tissues MMP production is regulated by a number of mechanisms including expression of the glycoprotein CD147. Previously, we have shown that CD147 is expressed on hepatocytes but not within the fibrotic septa in cirrhosis [1]. Therefore, we investigated if hepatocytes produce MMPs, regulated by CD147, which are capable of remodelling fibrotic ECM independent of the HSC.

Methods: Non-diseased, fibrotic and cirrhotic livers were examined for MMP activity and markers of fibrosis in humans and mice. CD147 expression and MMP activity were co-localised by in-situ zymography. The role of CD147 was studied in-vitro with siRNA to CD147 in hepatocytes and in-vivo in mice with CCl4 induced liver injury using ãCD147 antibody intervention.

Results: In liver fibrosis in both human and mouse tissue MMP expression and activity (MMP-2, -9, -13 and -14) increased with progressive injury and localised to hepatocytes. Additionally, as expected, MMPs were abundantly expressed by activated HSC. Further, with progressive fibrosis there was expression of CD147, which localised to hepatocytes but not to HSC. Functionally significant in-vitro regulation of hepatocyte MMP production by CD147 was demonstrated using siRNA to CD147 that decreased hepatocyte MMP-2 and -9 expression/activity. Further, in-vivo α-CD147 antibody intervention decreased liver MMP-2, -9, -13, -14, TGF-β and α-SMA expression in CCl4 treated mice compared to controls.

Conclusion: We have shown that hepatocytes produce active MMPs and that the glycoprotein CD147 regulates hepatocyte MMP expression. Targeting CD147 regulates hepatocyte MMP production both in-vitro and in-vivo, with the net result being reduced fibrotic matrix turnover in-vivo. Therefore, CD147 regulation of hepatocyte MMP is a novel pathway that could be targeted by future anti-fibrogenic agents.

Show MeSH
Related in: MedlinePlus