Adventitial transduction of lentivirus-shRNA-VEGF-A in arteriovenous fistula reduces venous stenosis formation

Binxia Yang, Rajiv Janardhanan, Pawan Vohra, Eddie L. Greene, Santanu Bhattacharya, Sarah Withers, Bhaskar Roy, Evelyn C. Nieves Torres, Jayawant Mandrekar, Edward B Leof, Debabrata Mukhopadhyay, Sanjay Misra

Research output: Contribution to journalArticle

30 Citations (Scopus)

Abstract

Venous neointimal hyperplasia (VNH) causes hemodialysis vascular access failure. Here we tested whether VNH formation occurs in part due to local vessel hypoxia caused by surgical trauma to the vasa vasorum of the outflow vein at the time of arteriovenous fistula placement. Selective targeting of the adventitia of the outflow vein at the time of fistula creation was performed using a lentivirus-delivered small-hairpin RNA that inhibits VEGF-A expression. This resulted in significant increase in mean lumen vessel area, decreased media/adventitia area, and decreased constrictive remodeling with a significant increase in apoptosis (increase in caspase 3 activity and TUNEL staining) accompanied with decreased cellular proliferation and hypoxia-inducible factor-1α at the outflow vein. There was significant decrease in cells staining positive for α-smooth muscle actin (a myofibroblast marker) and VEGFR-1 expression with a decrease in MMP-2 and MMP-9. These results were confirmed in animals that were treated with humanized monoclonal antibody to VEGF-A with similar results. Since hypoxia can cause fibroblast to differentiate into myofibroblasts, we silenced VEGF-A gene expression in fibroblasts and subjected them to hypoxia. This decreased myofibroblast production, cellular proliferation, cell invasion, MMP-2 activity, and increased caspase 3. Thus, VEGF-A reduction at the time of arteriovenous fistula placement results in increased positive vascular remodeling.

Original languageEnglish (US)
Pages (from-to)289-306
Number of pages18
JournalKidney International
Volume85
Issue number2
DOIs
StatePublished - Feb 2014

Fingerprint

Adventitia
Lentivirus
Arteriovenous Fistula
Vascular Endothelial Growth Factor A
Small Interfering RNA
Myofibroblasts
Pathologic Constriction
Matrix Metalloproteinases
Veins
Caspase 3
Hyperplasia
Fibroblasts
Vasa Vasorum
Cell Proliferation
Staining and Labeling
Vascular Endothelial Growth Factor Receptor-1
Antibodies, Monoclonal, Humanized
Hypoxia-Inducible Factor 1
Cell Hypoxia
In Situ Nick-End Labeling

Keywords

  • diabetes
  • vascular access
  • VEGF

ASJC Scopus subject areas

  • Nephrology

Cite this

Adventitial transduction of lentivirus-shRNA-VEGF-A in arteriovenous fistula reduces venous stenosis formation. / Yang, Binxia; Janardhanan, Rajiv; Vohra, Pawan; Greene, Eddie L.; Bhattacharya, Santanu; Withers, Sarah; Roy, Bhaskar; Nieves Torres, Evelyn C.; Mandrekar, Jayawant; Leof, Edward B; Mukhopadhyay, Debabrata; Misra, Sanjay.

In: Kidney International, Vol. 85, No. 2, 02.2014, p. 289-306.

Research output: Contribution to journalArticle

Yang, Binxia ; Janardhanan, Rajiv ; Vohra, Pawan ; Greene, Eddie L. ; Bhattacharya, Santanu ; Withers, Sarah ; Roy, Bhaskar ; Nieves Torres, Evelyn C. ; Mandrekar, Jayawant ; Leof, Edward B ; Mukhopadhyay, Debabrata ; Misra, Sanjay. / Adventitial transduction of lentivirus-shRNA-VEGF-A in arteriovenous fistula reduces venous stenosis formation. In: Kidney International. 2014 ; Vol. 85, No. 2. pp. 289-306.
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