Hepatocyte growth factor/c-met signaling is required for stem-cell-mediated liver regeneration in mice

Tsuyoshi Ishikawa, Valentina M. Factor, Jens U. Marquardt, Chiara Raggi, Daekwan Seo, Mitsuteru Kitade, Elizabeth A. Conner, Snorri S. Thorgeirsson

Research output: Contribution to journalArticle

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Abstract

Hepatocyte growth factor (HGF)/c-Met supports a pleiotrophic signal transduction pathway that controls stem cell homeostasis. Here, we directly addressed the role of c-Met in stem-cell-mediated liver regeneration by utilizing mice harboring c-met floxed alleles and Alb-Cre or Mx1-Cre transgenes. To activate oval cells, the hepatic stem cell (HSC) progeny, we used a model of liver injury induced by diet containing the porphyrinogenic agent, 3,5-diethocarbonyl-1,4-dihydrocollidine (DDC). Deletion of c-met in oval cells was confirmed in both models by polymerase chain reaction analysis of fluorescence-activated cell-sorted epithelial cell adhesion molecule (EpCam)-positive cells. Loss of c-Met receptor decreased the sphere-forming capacity of oval cells in vitro as well as reduced oval cell pool, impaired migration, and decreased hepatocytic differentiation in vivo, as demonstrated by double immunofluorescence using oval- (A6 and EpCam) and hepatocyte-specific (i.e. hepatocyte nuclear factor 4-alpha) antibodies. Furthermore, lack of c-Met had a profound effect on tissue remodeling and overall composition of HSC niche, which was associated with greatly reduced matrix metalloproteinase (MMP)9 activity and decreased expression of stromal-cell-derived factor 1. Using a combination of double immunofluorescence of cell-type-specific markers with MMP9 and gelatin zymography on the isolated cell populations, we identified macrophages as a major source of MMP9 in DDC-treated livers. The Mx1-Cre-driven c-met deletion caused the greatest phenotypic impact on HSCs response, as compared to the selective inactivation in the epithelial cell lineages achieved in c-Met fl/fl; Alb-Cre +/- mice. However, in both models, genetic loss of c-met triggered a similar cascade of events, leading to the failure of HSC mobilization and death of the mice. Conclusion: These results establish a direct contribution of c-Met in the regulation of HSC response and support a unique role for HGF/c-Met as an essential growth-factor-signaling pathway for regeneration of diseased liver.

Original languageEnglish
Pages (from-to)1215-1226
Number of pages12
JournalHepatology
Volume55
Issue number4
DOIs
Publication statusPublished - Apr 2012

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Liver Regeneration
Hepatocyte Growth Factor
Stem Cells
Hepatocytes
Fluorescent Antibody Technique
Hepatocyte Nuclear Factor 4
Hematopoietic Stem Cell Mobilization
Stem Cell Niche
Chemokine CXCL12
Liver
Genetic Models
Matrix Metalloproteinase 9
Cell Lineage
Gelatin
Transgenes
Liver Diseases
Signal Transduction
Intercellular Signaling Peptides and Proteins
Homeostasis
Cell Death

ASJC Scopus subject areas

  • Hepatology

Cite this

Ishikawa, T., Factor, V. M., Marquardt, J. U., Raggi, C., Seo, D., Kitade, M., ... Thorgeirsson, S. S. (2012). Hepatocyte growth factor/c-met signaling is required for stem-cell-mediated liver regeneration in mice. Hepatology, 55(4), 1215-1226. https://doi.org/10.1002/hep.24796

Hepatocyte growth factor/c-met signaling is required for stem-cell-mediated liver regeneration in mice. / Ishikawa, Tsuyoshi; Factor, Valentina M.; Marquardt, Jens U.; Raggi, Chiara; Seo, Daekwan; Kitade, Mitsuteru; Conner, Elizabeth A.; Thorgeirsson, Snorri S.

In: Hepatology, Vol. 55, No. 4, 04.2012, p. 1215-1226.

Research output: Contribution to journalArticle

Ishikawa, T, Factor, VM, Marquardt, JU, Raggi, C, Seo, D, Kitade, M, Conner, EA & Thorgeirsson, SS 2012, 'Hepatocyte growth factor/c-met signaling is required for stem-cell-mediated liver regeneration in mice', Hepatology, vol. 55, no. 4, pp. 1215-1226. https://doi.org/10.1002/hep.24796
Ishikawa, Tsuyoshi ; Factor, Valentina M. ; Marquardt, Jens U. ; Raggi, Chiara ; Seo, Daekwan ; Kitade, Mitsuteru ; Conner, Elizabeth A. ; Thorgeirsson, Snorri S. / Hepatocyte growth factor/c-met signaling is required for stem-cell-mediated liver regeneration in mice. In: Hepatology. 2012 ; Vol. 55, No. 4. pp. 1215-1226.
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