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Hepatocyte growth factor gene therapy enhances infiltration of macrophages and may induce kidney repair in db/db mice as a model of diabetes.

Flaquer M, Franquesa M, Vidal A, Bolaños N, Torras J, Lloberas N, Herrero-Fresneda I, Grinyó JM, Cruzado JM - Diabetologia (2012)

Bottom Line: The aim of this study was to test whether bone-marrow-derived cells are also involved in this HGF-induced reparative process.We have created chimeric db/db mice as a model of diabetes that produce enhanced green fluorescent protein (EGFP) in bone marrow cells.These cells are mainly monocyte-derived macrophages, which may contribute to the renal tissue repair and regeneration consistently observed in our model.

View Article: PubMed Central - PubMed

Affiliation: Nephrology Laboratory, Departament de Ciències Clíniques, IDIBELL, University of Barcelona, Bellvitge Hospital, Barcelona, Spain.

ABSTRACT

Aims/hypothesis: We previously demonstrated hepatocyte growth factor (HGF) gene therapy was able to induce regression of glomerulosclerosis in diabetic nephropathy through local reparative mechanisms. The aim of this study was to test whether bone-marrow-derived cells are also involved in this HGF-induced reparative process.

Methods: We have created chimeric db/db mice as a model of diabetes that produce enhanced green fluorescent protein (EGFP) in bone marrow cells. We performed treatment with HGF gene therapy either alone or in combination with granulocyte-colony stimulating factor, in order to induce mobilisation of haematopoietic stem cells in these diabetic and chimeric animals.

Results: We find HGF gene therapy enhances renal expression of stromal-cell-derived factor-1 and is subsequently associated with an increased number of bone-marrow-derived cells getting into the injured kidneys. These cells are mainly monocyte-derived macrophages, which may contribute to the renal tissue repair and regeneration consistently observed in our model. Finally, HGF gene therapy is associated with the presence of a small number of Bowman's capsule parietal epithelial cells producing EGFP, suggesting they are fused with bone-marrow-derived cells and are contributing to podocyte repopulation.

Conclusions/interpretation: Altogether, our findings provide new evidence about the therapeutic role of HGF and open new opportunities for inducing renal regeneration in diabetic nephropathy.

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Effect of HGF gene therapy on diabetic kidney disease. a Collagen IV immunofluorescence (×630). Only HGF gene therapy (either alone or in combination with G-CSF) was associated with a significant decrease in collagen IV. b Fibronectin immunohistochemistry (×630). Groups treated with HGF therapy (db/db+HGF and db/db+HGF+G-CSF) showed decreased fibronectin accumulation. c Histogram summarising the mean values of collagen IV and fibronectin from each group. Black bars, collagen IV; white bars, fibronectin *p < 0.05 vs db/db, db/db–BMT and db/db+G-CSF; †p < 0.05 vs db/db+HGF and db/db+HGF+G-CSF (Kruskal–Wallis and Mann–Whitney U tests)
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Fig2: Effect of HGF gene therapy on diabetic kidney disease. a Collagen IV immunofluorescence (×630). Only HGF gene therapy (either alone or in combination with G-CSF) was associated with a significant decrease in collagen IV. b Fibronectin immunohistochemistry (×630). Groups treated with HGF therapy (db/db+HGF and db/db+HGF+G-CSF) showed decreased fibronectin accumulation. c Histogram summarising the mean values of collagen IV and fibronectin from each group. Black bars, collagen IV; white bars, fibronectin *p < 0.05 vs db/db, db/db–BMT and db/db+G-CSF; †p < 0.05 vs db/db+HGF and db/db+HGF+G-CSF (Kruskal–Wallis and Mann–Whitney U tests)

Mentions: Baseline and post-intervention functional variables are summarised in Table 1. Glucose levels remained similar in diabetic-treated and non-treated groups, suggesting that neither HGF nor G-CSF treatments have any effect on hyperglycaemia. Nevertheless, HGF gene therapy and/or G-CSF administration halted progression of albuminuria. We further assessed whether these therapies had a role in the progression of glomerulosclerosis (Fig. 2a, b). The animals receiving HGF either alone or in combination with G-CSF showed significant reduction in both collagen IV and fibronectin glomerular deposition. Isolated G-CSF therapy was not associated with any significant reduction of glomerulosclerosis. Accordingly, HGF gene therapy rather than G-CSF reduced the Tgf-β/Hgf mRNA ratio in renal tissue (Fig. 3).Fig. 2


Hepatocyte growth factor gene therapy enhances infiltration of macrophages and may induce kidney repair in db/db mice as a model of diabetes.

Flaquer M, Franquesa M, Vidal A, Bolaños N, Torras J, Lloberas N, Herrero-Fresneda I, Grinyó JM, Cruzado JM - Diabetologia (2012)

Effect of HGF gene therapy on diabetic kidney disease. a Collagen IV immunofluorescence (×630). Only HGF gene therapy (either alone or in combination with G-CSF) was associated with a significant decrease in collagen IV. b Fibronectin immunohistochemistry (×630). Groups treated with HGF therapy (db/db+HGF and db/db+HGF+G-CSF) showed decreased fibronectin accumulation. c Histogram summarising the mean values of collagen IV and fibronectin from each group. Black bars, collagen IV; white bars, fibronectin *p < 0.05 vs db/db, db/db–BMT and db/db+G-CSF; †p < 0.05 vs db/db+HGF and db/db+HGF+G-CSF (Kruskal–Wallis and Mann–Whitney U tests)
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Related In: Results  -  Collection

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Fig2: Effect of HGF gene therapy on diabetic kidney disease. a Collagen IV immunofluorescence (×630). Only HGF gene therapy (either alone or in combination with G-CSF) was associated with a significant decrease in collagen IV. b Fibronectin immunohistochemistry (×630). Groups treated with HGF therapy (db/db+HGF and db/db+HGF+G-CSF) showed decreased fibronectin accumulation. c Histogram summarising the mean values of collagen IV and fibronectin from each group. Black bars, collagen IV; white bars, fibronectin *p < 0.05 vs db/db, db/db–BMT and db/db+G-CSF; †p < 0.05 vs db/db+HGF and db/db+HGF+G-CSF (Kruskal–Wallis and Mann–Whitney U tests)
Mentions: Baseline and post-intervention functional variables are summarised in Table 1. Glucose levels remained similar in diabetic-treated and non-treated groups, suggesting that neither HGF nor G-CSF treatments have any effect on hyperglycaemia. Nevertheless, HGF gene therapy and/or G-CSF administration halted progression of albuminuria. We further assessed whether these therapies had a role in the progression of glomerulosclerosis (Fig. 2a, b). The animals receiving HGF either alone or in combination with G-CSF showed significant reduction in both collagen IV and fibronectin glomerular deposition. Isolated G-CSF therapy was not associated with any significant reduction of glomerulosclerosis. Accordingly, HGF gene therapy rather than G-CSF reduced the Tgf-β/Hgf mRNA ratio in renal tissue (Fig. 3).Fig. 2

Bottom Line: The aim of this study was to test whether bone-marrow-derived cells are also involved in this HGF-induced reparative process.We have created chimeric db/db mice as a model of diabetes that produce enhanced green fluorescent protein (EGFP) in bone marrow cells.These cells are mainly monocyte-derived macrophages, which may contribute to the renal tissue repair and regeneration consistently observed in our model.

View Article: PubMed Central - PubMed

Affiliation: Nephrology Laboratory, Departament de Ciències Clíniques, IDIBELL, University of Barcelona, Bellvitge Hospital, Barcelona, Spain.

ABSTRACT

Aims/hypothesis: We previously demonstrated hepatocyte growth factor (HGF) gene therapy was able to induce regression of glomerulosclerosis in diabetic nephropathy through local reparative mechanisms. The aim of this study was to test whether bone-marrow-derived cells are also involved in this HGF-induced reparative process.

Methods: We have created chimeric db/db mice as a model of diabetes that produce enhanced green fluorescent protein (EGFP) in bone marrow cells. We performed treatment with HGF gene therapy either alone or in combination with granulocyte-colony stimulating factor, in order to induce mobilisation of haematopoietic stem cells in these diabetic and chimeric animals.

Results: We find HGF gene therapy enhances renal expression of stromal-cell-derived factor-1 and is subsequently associated with an increased number of bone-marrow-derived cells getting into the injured kidneys. These cells are mainly monocyte-derived macrophages, which may contribute to the renal tissue repair and regeneration consistently observed in our model. Finally, HGF gene therapy is associated with the presence of a small number of Bowman's capsule parietal epithelial cells producing EGFP, suggesting they are fused with bone-marrow-derived cells and are contributing to podocyte repopulation.

Conclusions/interpretation: Altogether, our findings provide new evidence about the therapeutic role of HGF and open new opportunities for inducing renal regeneration in diabetic nephropathy.

Show MeSH
Related in: MedlinePlus