Acute podocyte vascular endothelial growth factor (VEGF-A) knockdown disrupts alphavbeta3 integrin signaling in the glomerulus

de lma Veron, Guillermo Villegas, Pardeep Kumar Aggarwal, Claudia Bertuccio, Juan Jimenez, Heino Velazquez, Kimberly J. Reidy, Dale R. Abrahamson, Gilbert Moeckel, Michael Kashgarian, Alda Tufro

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Abstract

Podocyte or endothelial cell VEGF-A knockout causes thrombotic microangiopathy in adult mice. To study the mechanism involved in acute and local injury caused by low podocyte VEGF-A we developed an inducible, podocyte-specific VEGF-A knockdown mouse, and we generated an immortalized podocyte cell line (VEGFKD) that downregulates VEGF-A upon doxycycline exposure. Tet-O-siVEGF:podocin-rtTA mice express VEGF shRNA in podocytes in a doxycycline-regulated manner, decreasing VEGF-A mRNA and VEGF-A protein levels in isolated glomeruli to ~20% of non-induced controls and urine VEGF-A to ~30% of control values a week after doxycycline induction. Induced tet-O-siVEGF:podocin-rtTA mice developed acute renal failure and proteinuria, associated with mesangiolysis and microaneurisms. Glomerular ultrastructure revealed endothelial cell swelling, GBM lamination and podocyte effacement. VEGF knockdown decreased podocyte fibronectin and glomerular endothelial alphaVbeta3 integrin in vivo. VEGF receptor-2 (VEGFR2) interacts with beta3 integrin and neuropilin-1 in the kidney in vivo and in VEGFKD podocytes. Podocyte VEGF knockdown disrupts alphaVbeta3 integrin activation in glomeruli, detected by WOW1-Fab. VEGF silencing in cultured VEGFKD podocytes downregulates fibronectin and disrupts alphaVbeta3 integrin activation cell-autonomously. Collectively, these studies indicate that podocyte VEGF-A regulates alphaVbeta3 integrin signaling in the glomerulus, and that podocyte VEGF knockdown disrupts alphaVbeta3 integrin activity via decreased VEGFR2 signaling, thereby damaging the three layers of the glomerular filtration barrier, causing proteinuria and acute renal failure.

Original languageEnglish (US)
Article numbere40589
JournalPLoS One
Volume7
Issue number7
DOIs
StatePublished - Jul 13 2012

Fingerprint

Integrin alphaVbeta3
Podocytes
vascular endothelial growth factors
integrins
Vascular Endothelial Growth Factor A
doxycycline
vascular endothelial growth factor receptors
fibronectins
mice
renal failure
Doxycycline
endothelial cells
microangiopathy
Vascular Endothelial Growth Factor Receptor
Endothelial cells
Proteinuria
Fibronectins
Acute Kidney Injury
Down-Regulation
Glomerular Filtration Barrier

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Veron, D. L., Villegas, G., Aggarwal, P. K., Bertuccio, C., Jimenez, J., Velazquez, H., ... Tufro, A. (2012). Acute podocyte vascular endothelial growth factor (VEGF-A) knockdown disrupts alphavbeta3 integrin signaling in the glomerulus. PLoS One, 7(7), [e40589]. https://doi.org/10.1371/journal.pone.0040589

Acute podocyte vascular endothelial growth factor (VEGF-A) knockdown disrupts alphavbeta3 integrin signaling in the glomerulus. / Veron, de lma; Villegas, Guillermo; Aggarwal, Pardeep Kumar; Bertuccio, Claudia; Jimenez, Juan; Velazquez, Heino; Reidy, Kimberly J.; Abrahamson, Dale R.; Moeckel, Gilbert; Kashgarian, Michael; Tufro, Alda.

In: PLoS One, Vol. 7, No. 7, e40589, 13.07.2012.

Research output: Contribution to journalArticle

Veron, DL, Villegas, G, Aggarwal, PK, Bertuccio, C, Jimenez, J, Velazquez, H, Reidy, KJ, Abrahamson, DR, Moeckel, G, Kashgarian, M & Tufro, A 2012, 'Acute podocyte vascular endothelial growth factor (VEGF-A) knockdown disrupts alphavbeta3 integrin signaling in the glomerulus', PLoS One, vol. 7, no. 7, e40589. https://doi.org/10.1371/journal.pone.0040589
Veron, de lma ; Villegas, Guillermo ; Aggarwal, Pardeep Kumar ; Bertuccio, Claudia ; Jimenez, Juan ; Velazquez, Heino ; Reidy, Kimberly J. ; Abrahamson, Dale R. ; Moeckel, Gilbert ; Kashgarian, Michael ; Tufro, Alda. / Acute podocyte vascular endothelial growth factor (VEGF-A) knockdown disrupts alphavbeta3 integrin signaling in the glomerulus. In: PLoS One. 2012 ; Vol. 7, No. 7.
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abstract = "Podocyte or endothelial cell VEGF-A knockout causes thrombotic microangiopathy in adult mice. To study the mechanism involved in acute and local injury caused by low podocyte VEGF-A we developed an inducible, podocyte-specific VEGF-A knockdown mouse, and we generated an immortalized podocyte cell line (VEGFKD) that downregulates VEGF-A upon doxycycline exposure. Tet-O-siVEGF:podocin-rtTA mice express VEGF shRNA in podocytes in a doxycycline-regulated manner, decreasing VEGF-A mRNA and VEGF-A protein levels in isolated glomeruli to ~20{\%} of non-induced controls and urine VEGF-A to ~30{\%} of control values a week after doxycycline induction. Induced tet-O-siVEGF:podocin-rtTA mice developed acute renal failure and proteinuria, associated with mesangiolysis and microaneurisms. Glomerular ultrastructure revealed endothelial cell swelling, GBM lamination and podocyte effacement. VEGF knockdown decreased podocyte fibronectin and glomerular endothelial alphaVbeta3 integrin in vivo. VEGF receptor-2 (VEGFR2) interacts with beta3 integrin and neuropilin-1 in the kidney in vivo and in VEGFKD podocytes. Podocyte VEGF knockdown disrupts alphaVbeta3 integrin activation in glomeruli, detected by WOW1-Fab. VEGF silencing in cultured VEGFKD podocytes downregulates fibronectin and disrupts alphaVbeta3 integrin activation cell-autonomously. Collectively, these studies indicate that podocyte VEGF-A regulates alphaVbeta3 integrin signaling in the glomerulus, and that podocyte VEGF knockdown disrupts alphaVbeta3 integrin activity via decreased VEGFR2 signaling, thereby damaging the three layers of the glomerular filtration barrier, causing proteinuria and acute renal failure.",
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AU - Velazquez, Heino

AU - Reidy, Kimberly J.

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