Limits...
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.

Show MeSH

Related in: MedlinePlus

Evaluation of renal EGFP+ cells. a Mean values of automatic EGFP quantification in renal tissue. *p < 0.02 vs db/db+G-CSF and db/db+HGF; **p < 0.01 vs db/db–BMT, db/db+G-CSF and db/db+HGF (Mann–Whitney U test). b Representative sample of renal tissue from db/db+HGF+G-CSF group (×40) showing cortical (arrowhead) as well as medullar (arrow) EGFP+ cells. c Glomerular detail (×630) using CD34 staining (red) in order to identify endothelial cells. The asterisk shows a glomerular extravascular EGFP+ cell and the arrowhead shows a circulating EGFP+ cell
© Copyright Policy
Related In: Results  -  Collection


getmorefigures.php?uid=PMC3369134&req=5

Fig5: Evaluation of renal EGFP+ cells. a Mean values of automatic EGFP quantification in renal tissue. *p < 0.02 vs db/db+G-CSF and db/db+HGF; **p < 0.01 vs db/db–BMT, db/db+G-CSF and db/db+HGF (Mann–Whitney U test). b Representative sample of renal tissue from db/db+HGF+G-CSF group (×40) showing cortical (arrowhead) as well as medullar (arrow) EGFP+ cells. c Glomerular detail (×630) using CD34 staining (red) in order to identify endothelial cells. The asterisk shows a glomerular extravascular EGFP+ cell and the arrowhead shows a circulating EGFP+ cell

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)

Evaluation of renal EGFP+ cells. a Mean values of automatic EGFP quantification in renal tissue. *p < 0.02 vs db/db+G-CSF and db/db+HGF; **p < 0.01 vs db/db–BMT, db/db+G-CSF and db/db+HGF (Mann–Whitney U test). b Representative sample of renal tissue from db/db+HGF+G-CSF group (×40) showing cortical (arrowhead) as well as medullar (arrow) EGFP+ cells. c Glomerular detail (×630) using CD34 staining (red) in order to identify endothelial cells. The asterisk shows a glomerular extravascular EGFP+ cell and the arrowhead shows a circulating EGFP+ cell
© Copyright Policy
Related In: Results  -  Collection

Show All Figures
getmorefigures.php?uid=PMC3369134&req=5

Fig5: Evaluation of renal EGFP+ cells. a Mean values of automatic EGFP quantification in renal tissue. *p < 0.02 vs db/db+G-CSF and db/db+HGF; **p < 0.01 vs db/db–BMT, db/db+G-CSF and db/db+HGF (Mann–Whitney U test). b Representative sample of renal tissue from db/db+HGF+G-CSF group (×40) showing cortical (arrowhead) as well as medullar (arrow) EGFP+ cells. c Glomerular detail (×630) using CD34 staining (red) in order to identify endothelial cells. The asterisk shows a glomerular extravascular EGFP+ cell and the arrowhead shows a circulating EGFP+ cell
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