Megan Stevens1,2,3, Christopher R Neal2,3, Andrew H J Salmon2,3, David O Bates4, Steven J Harper2,3, Sebastian Oltean1,2,3. 1. Institute of Biomedical and Clinical Sciences, University of Exeter Medical School, Exeter, United Kingdom. 2. School of Physiology, Pharmacology and Neurosciences, Bristol, United Kingdom. 3. Bristol Renal, School of Clinical Sciences, University of Bristol, Bristol, United Kingdom. 4. Division of Cancer and Stem Cells, School of Medicine, University of Nottingham, Queen's Medical Centre, Nottingham, United Kingdom.
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.
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.
Authors: M Saito; T Iwawaki; C Taya; H Yonekawa; M Noda; Y Inui; E Mekada; Y Kimata; A Tsuru; K Kohno Journal: Nat Biotechnol Date: 2001-08 Impact factor: 54.908
Authors: Jian-Kan Guo; Arnaud Marlier; Hongmei Shi; Alan Shan; Thomas A Ardito; Zhao-Peng Du; Michael Kashgarian; Diane S Krause; Daniel Biemesderfer; Lloyd G Cantley Journal: J Am Soc Nephrol Date: 2011-12-22 Impact factor: 10.121
Authors: Bart Smeets; Henry B P M Dijkman; Nathalie A J M te Loeke; Jacco P H F van Son; Eric J Steenbergen; Karel J M Assmann; Jack F M Wetzels; Patricia J T A Groenen Journal: Nephrol Dial Transplant Date: 2003-12 Impact factor: 5.992
Authors: Yan Qiu; Joanne Ferguson; Sebastian Oltean; Chris R Neal; Amit Kaura; Heather Bevan; Emma Wood; Leslie M Sage; Silvia Lanati; Dawid G Nowak; Andy H J Salmon; David Bates; Steve J Harper Journal: J Am Soc Nephrol Date: 2010-08-05 Impact factor: 10.121
Authors: Natalie C Finch; Sarah S Fawaz; Chris R Neal; Matthew J Butler; Vivian K Lee; Andrew J Salmon; Abigail C Lay; Megan Stevens; Lusyan Dayalan; Hamid Band; Harry H Mellor; Steven J Harper; David T Shima; Gavin I Welsh; Rebecca R Foster; Simon C Satchell Journal: J Am Soc Nephrol Date: 2022-03-15 Impact factor: 14.978