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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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Related in: MedlinePlus

HSC mobilisation and SDF-1 renal expression. a Peripheral blood HSC mobilisation. Only the group receiving G-CSF had increased circulating HSC, which was maximal at days 8 and 22 (2 days after each G-CSF injection). HGF had no effect on HSC mobilisation. Diamonds, db/db–BMT; squares, db/db+HGF; triangles, db/db+HGF+G-CSF. b SDF-1 renal staining: db/db–BMT; db/db+G-CSF; db/db+HGF+G-CSF; and db/db+HGF. Animals treated with HGF displayed higher production of SDF-1 in comparison with those without HGF treatment. c Graph illustrating the mean values of SDF-1 production in different treatment groups. *p < 0.05 vs db/db+HGF and db/db–BMT+HGF+G-CSF (Mann–Whitney U test)
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Fig4: HSC mobilisation and SDF-1 renal expression. a Peripheral blood HSC mobilisation. Only the group receiving G-CSF had increased circulating HSC, which was maximal at days 8 and 22 (2 days after each G-CSF injection). HGF had no effect on HSC mobilisation. Diamonds, db/db–BMT; squares, db/db+HGF; triangles, db/db+HGF+G-CSF. b SDF-1 renal staining: db/db–BMT; db/db+G-CSF; db/db+HGF+G-CSF; and db/db+HGF. Animals treated with HGF displayed higher production of SDF-1 in comparison with those without HGF treatment. c Graph illustrating the mean values of SDF-1 production in different treatment groups. *p < 0.05 vs db/db+HGF and db/db–BMT+HGF+G-CSF (Mann–Whitney U test)

Mentions: We analysed whether HGF induced HSC mobilisation into peripheral blood (Fig. 4a). No mobilisation was observed when animals received HGF (db/db–BMT+HGF). The addition of G-CSF to these animals significantly increased the number of circulating HSCs. We found HGF supplementation was associated with enhanced SDF-1 production in renal tissue (Fig. 4b, c). However, mobilisation of bone marrow cells by administration of G-CSF led to an increased number of these cells in the kidney, as shown in Fig. 5. Administration of HGF also led to an increase in renal EGFP+ fluorescence, conceivably because of its ability to induce SDF-1 production in renal tissue. The combination of HGF gene therapy with HSC mobilisation by G-CSF robustly increased the amount of bone-marrow-derived EGFP+ cells in the diabetic kidney (Fig. 5a). We found bone-marrow-derived cells in the renal cortex and medulla (Fig. 5b). Some of these EGFP+ cells were localised in diabetic glomeruli (Fig. 5c).Fig. 4


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)

HSC mobilisation and SDF-1 renal expression. a Peripheral blood HSC mobilisation. Only the group receiving G-CSF had increased circulating HSC, which was maximal at days 8 and 22 (2 days after each G-CSF injection). HGF had no effect on HSC mobilisation. Diamonds, db/db–BMT; squares, db/db+HGF; triangles, db/db+HGF+G-CSF. b SDF-1 renal staining: db/db–BMT; db/db+G-CSF; db/db+HGF+G-CSF; and db/db+HGF. Animals treated with HGF displayed higher production of SDF-1 in comparison with those without HGF treatment. c Graph illustrating the mean values of SDF-1 production in different treatment groups. *p < 0.05 vs db/db+HGF and db/db–BMT+HGF+G-CSF (Mann–Whitney U test)
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Related In: Results  -  Collection

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Fig4: HSC mobilisation and SDF-1 renal expression. a Peripheral blood HSC mobilisation. Only the group receiving G-CSF had increased circulating HSC, which was maximal at days 8 and 22 (2 days after each G-CSF injection). HGF had no effect on HSC mobilisation. Diamonds, db/db–BMT; squares, db/db+HGF; triangles, db/db+HGF+G-CSF. b SDF-1 renal staining: db/db–BMT; db/db+G-CSF; db/db+HGF+G-CSF; and db/db+HGF. Animals treated with HGF displayed higher production of SDF-1 in comparison with those without HGF treatment. c Graph illustrating the mean values of SDF-1 production in different treatment groups. *p < 0.05 vs db/db+HGF and db/db–BMT+HGF+G-CSF (Mann–Whitney U test)
Mentions: We analysed whether HGF induced HSC mobilisation into peripheral blood (Fig. 4a). No mobilisation was observed when animals received HGF (db/db–BMT+HGF). The addition of G-CSF to these animals significantly increased the number of circulating HSCs. We found HGF supplementation was associated with enhanced SDF-1 production in renal tissue (Fig. 4b, c). However, mobilisation of bone marrow cells by administration of G-CSF led to an increased number of these cells in the kidney, as shown in Fig. 5. Administration of HGF also led to an increase in renal EGFP+ fluorescence, conceivably because of its ability to induce SDF-1 production in renal tissue. The combination of HGF gene therapy with HSC mobilisation by G-CSF robustly increased the amount of bone-marrow-derived EGFP+ cells in the diabetic kidney (Fig. 5a). We found bone-marrow-derived cells in the renal cortex and medulla (Fig. 5b). Some of these EGFP+ cells were localised in diabetic glomeruli (Fig. 5c).Fig. 4

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