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Therapeutic effect of hepatocyte growth factor-secreting mesenchymal stem cells in a rat model of liver fibrosis.

Kim MD, Kim SS, Cha HY, Jang SH, Chang DY, Kim W, Suh-Kim H, Lee JH - Exp. Mol. Med. (2014)

Bottom Line: Assessment of cell and biochemical parameters revealed that mRNA levels of the fibrogenic cytokines PDGF-bb and TGF-β1 were significantly decreased after MSC/HGF therapy.Subsequent to the decrease in collagen, expression of matrix metalloprotease-9 (MMP-9), MMP-13, MMP-14 and urokinase-type plasminogen activator was augmented following MSC/HGF, whereas tissue inhibitor of metalloprotease-1 (TIMP-1) expression was reduced.In conclusion, therapy with MSCs/HGF resulted in an improved therapeutic effect compared with MSCs alone, probably because of the anti-fibrotic activity of HGF.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biochemistry, Ajou University School of Medicine, Suwon, Republic of Korea [2] Chronic Inflammatory Disease Research Center, Ajou University School of Medicine, Suwon, Republic of Korea.

ABSTRACT
Bone marrow-derived mesenchymal stromal cells (MSCs) have been reported to be beneficial for the treatment of liver fibrosis. Here, we investigated the use of genetically engineered MSCs that overexpress hepatocyte growth factor (HGF) as a means to improve their therapeutic effect in liver fibrosis. Liver fibrosis was induced by intraperitoneal injection of dimethylnitrosamine. HGF-secreting MSCs (MSCs/HGF) were prepared by transducing MSCs with an adenovirus carrying HGF-encoding cDNA. MSCs or MSCs/HGF were injected directly into the spleen of fibrotic rats. Tissue fibrosis was assessed by histological analysis 12 days after stem cell injection. Although treatment with MSCs reduced fibrosis, treatment with MSCs/HGF produced a more significant reduction and was associated with elevated HGF levels in the portal vein. Collagen levels in the liver extract were decreased after MSC/HGF therapy, suggesting recovery from fibrosis. Furthermore, liver function was improved in animals receiving MSCs/HGF, indicating that MSC/HGF therapy resulted not only in reduction of liver fibrosis but also in improvement of hepatocyte function. Assessment of cell and biochemical parameters revealed that mRNA levels of the fibrogenic cytokines PDGF-bb and TGF-β1 were significantly decreased after MSC/HGF therapy. Subsequent to the decrease in collagen, expression of matrix metalloprotease-9 (MMP-9), MMP-13, MMP-14 and urokinase-type plasminogen activator was augmented following MSC/HGF, whereas tissue inhibitor of metalloprotease-1 (TIMP-1) expression was reduced. In conclusion, therapy with MSCs/HGF resulted in an improved therapeutic effect compared with MSCs alone, probably because of the anti-fibrotic activity of HGF. Thus, MSC/HGF represents a promising approach toward a cell therapy for liver fibrosis.

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Analysis of HGF secreted by MSCs/HGF. (a) The amount of human HGF after 4 days of culture. (b) Secretion of HGF by MSCs or MSCs/HGF at 2, 4, 6, 8 and 10 days after Ad-HGF transduction. (c) Immunoblot showing c-Met phosphorylation in MDCK2 cells treated with conditioned media obtained from indicated cells. Human recombinant HGF protein (rhHGF, 100 unit ml−1) was used as a positive control. IB, immunoblotting; IP, immunoprecipitation; pY, phospho-tyrosine.
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fig1: Analysis of HGF secreted by MSCs/HGF. (a) The amount of human HGF after 4 days of culture. (b) Secretion of HGF by MSCs or MSCs/HGF at 2, 4, 6, 8 and 10 days after Ad-HGF transduction. (c) Immunoblot showing c-Met phosphorylation in MDCK2 cells treated with conditioned media obtained from indicated cells. Human recombinant HGF protein (rhHGF, 100 unit ml−1) was used as a positive control. IB, immunoblotting; IP, immunoprecipitation; pY, phospho-tyrosine.

Mentions: The amount of HGF produced by MSCs transduced with Ad-HGF was assessed by ELISA and was ∼160 times higher than the amount endogenously secreted by MSCs (557.86 vs 3.49 ng ml−1, from 5 × 105 cells cultured for 4 days, Figure 1a). HGF production peaked at 2–4 days after transduction and was maintained for at least 10 days (Figure 1b). The biological activity of HGF produced by MSCs/HGF was confirmed by phosphorylation of c-Met, the receptor for HGF (Figure 1c), and by scattering of MDCK2 cells (data not shown), a characteristic function of HGF in these cells.24


Therapeutic effect of hepatocyte growth factor-secreting mesenchymal stem cells in a rat model of liver fibrosis.

Kim MD, Kim SS, Cha HY, Jang SH, Chang DY, Kim W, Suh-Kim H, Lee JH - Exp. Mol. Med. (2014)

Analysis of HGF secreted by MSCs/HGF. (a) The amount of human HGF after 4 days of culture. (b) Secretion of HGF by MSCs or MSCs/HGF at 2, 4, 6, 8 and 10 days after Ad-HGF transduction. (c) Immunoblot showing c-Met phosphorylation in MDCK2 cells treated with conditioned media obtained from indicated cells. Human recombinant HGF protein (rhHGF, 100 unit ml−1) was used as a positive control. IB, immunoblotting; IP, immunoprecipitation; pY, phospho-tyrosine.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4150933&req=5

fig1: Analysis of HGF secreted by MSCs/HGF. (a) The amount of human HGF after 4 days of culture. (b) Secretion of HGF by MSCs or MSCs/HGF at 2, 4, 6, 8 and 10 days after Ad-HGF transduction. (c) Immunoblot showing c-Met phosphorylation in MDCK2 cells treated with conditioned media obtained from indicated cells. Human recombinant HGF protein (rhHGF, 100 unit ml−1) was used as a positive control. IB, immunoblotting; IP, immunoprecipitation; pY, phospho-tyrosine.
Mentions: The amount of HGF produced by MSCs transduced with Ad-HGF was assessed by ELISA and was ∼160 times higher than the amount endogenously secreted by MSCs (557.86 vs 3.49 ng ml−1, from 5 × 105 cells cultured for 4 days, Figure 1a). HGF production peaked at 2–4 days after transduction and was maintained for at least 10 days (Figure 1b). The biological activity of HGF produced by MSCs/HGF was confirmed by phosphorylation of c-Met, the receptor for HGF (Figure 1c), and by scattering of MDCK2 cells (data not shown), a characteristic function of HGF in these cells.24

Bottom Line: Assessment of cell and biochemical parameters revealed that mRNA levels of the fibrogenic cytokines PDGF-bb and TGF-β1 were significantly decreased after MSC/HGF therapy.Subsequent to the decrease in collagen, expression of matrix metalloprotease-9 (MMP-9), MMP-13, MMP-14 and urokinase-type plasminogen activator was augmented following MSC/HGF, whereas tissue inhibitor of metalloprotease-1 (TIMP-1) expression was reduced.In conclusion, therapy with MSCs/HGF resulted in an improved therapeutic effect compared with MSCs alone, probably because of the anti-fibrotic activity of HGF.

View Article: PubMed Central - PubMed

Affiliation: 1] Department of Biochemistry, Ajou University School of Medicine, Suwon, Republic of Korea [2] Chronic Inflammatory Disease Research Center, Ajou University School of Medicine, Suwon, Republic of Korea.

ABSTRACT
Bone marrow-derived mesenchymal stromal cells (MSCs) have been reported to be beneficial for the treatment of liver fibrosis. Here, we investigated the use of genetically engineered MSCs that overexpress hepatocyte growth factor (HGF) as a means to improve their therapeutic effect in liver fibrosis. Liver fibrosis was induced by intraperitoneal injection of dimethylnitrosamine. HGF-secreting MSCs (MSCs/HGF) were prepared by transducing MSCs with an adenovirus carrying HGF-encoding cDNA. MSCs or MSCs/HGF were injected directly into the spleen of fibrotic rats. Tissue fibrosis was assessed by histological analysis 12 days after stem cell injection. Although treatment with MSCs reduced fibrosis, treatment with MSCs/HGF produced a more significant reduction and was associated with elevated HGF levels in the portal vein. Collagen levels in the liver extract were decreased after MSC/HGF therapy, suggesting recovery from fibrosis. Furthermore, liver function was improved in animals receiving MSCs/HGF, indicating that MSC/HGF therapy resulted not only in reduction of liver fibrosis but also in improvement of hepatocyte function. Assessment of cell and biochemical parameters revealed that mRNA levels of the fibrogenic cytokines PDGF-bb and TGF-β1 were significantly decreased after MSC/HGF therapy. Subsequent to the decrease in collagen, expression of matrix metalloprotease-9 (MMP-9), MMP-13, MMP-14 and urokinase-type plasminogen activator was augmented following MSC/HGF, whereas tissue inhibitor of metalloprotease-1 (TIMP-1) expression was reduced. In conclusion, therapy with MSCs/HGF resulted in an improved therapeutic effect compared with MSCs alone, probably because of the anti-fibrotic activity of HGF. Thus, MSC/HGF represents a promising approach toward a cell therapy for liver fibrosis.

Show MeSH
Related in: MedlinePlus