Vascular endothelial growth factor-A165b restores normal glomerular water permeability in a diphtheria-toxin mouse model of glomerular injury

Stevens, Megan; Neal, Christopher R.; Salmon, Andrew H.J.; Bates, David O.; Harper, Steven J.; Oltean, Sebastian

Abstract

BACKGROUND/AIMS:Genetic cell ablation using the human diphtheria toxin receptor (hDTR) is a new strategy used for analysing cellular function. Diphtheria toxin (DT) is a cytotoxic protein that leaves mouse cells relatively unaffected, but upon binding to hDTR it ultimately leads to cell death. We used a podocyte-specific hDTR expressing (Pod-DTR) mouse to assess the anti-permeability and cyto-protective effects of the splice isoform vascular endothelial growth factor (VEGF-A165b).
METHODS:The Pod-DTR mouse was crossed with a mouse that over-expressed VEGF-A165b specifically in the podocytes (Neph-VEGF-A165b). Wild type (WT), Pod-DTR, Neph-VEGF-A165b and Pod-DTR X Neph-VEGF-A165b mice were treated with several doses of DT (1, 5, 100, and 1,000 ng/g bodyweight). Urine was collected and the glomerular water permeability (LpA/Vi) was measured ex vivo after 14 days. Structural analysis and podocyte marker expression were also assessed.
RESULTS: Pod-DTR mice developed an increased glomerular LpA/Vi 14 days after administration of DT (all doses), which was prevented when the mice over-expressed VEGF-A165b. No major structural abnormalities, podocyte ablation or albuminuria was observed in Pod-DTR mice, indicating this to be a mild model of podocyte disease. However, a change in expression and localisation of nephrin within the podocytes was observed, indicating disruption of the slit diaphragm in the Pod-DTR mice. This was prevented in the Pod-DTR X Neph-VEGF-A165b mice.
CONCLUSION: Although only a mild model of podocyte injury, over-expression of the anti-permeability VEGF-A165b isoform in the podocytes of Pod-DTR mice had a protective effect. Therefore, this study further highlights the therapeutic potential of VEGF-A165b in glomerular disease.

Citation

Stevens, M., Neal, C. R., Salmon, A. H., Bates, D. O., Harper, S. J., & Oltean, S. (in press). Vascular endothelial growth factor-A165b restores normal glomerular water permeability in a diphtheria-toxin mouse model of glomerular injury. Nephron, https://doi.org/10.1159/000485664

• Journal Article Type: Article
• Acceptance Date: Nov 24, 2017
• Online Publication Date: Jan 26, 2018
• Deposit Date: Mar 5, 2018
• Publicly Available Date: Mar 5, 2018
• Journal: Nephron
• Print ISSN: 0028-2766
• Electronic ISSN: 1423-0186
• Peer Reviewed: Peer Reviewed
• DOI: https://doi.org/10.1159/000485664
• Public URL: https://nottingham-repository.worktribe.com/output/907142
• Publisher URL: https://www.karger.com/Article/FullText/485664
• Contract Date: Mar 5, 2018