Adenovirus-based vascular endothelial growth factor gene delivery to human pancreatic islets

K. Cheng, D. Fraga, C. Zhang, M. Kotb, A. O. Gaber, Ramareddy Guntaka, R. I. Mahato

Research output: Contribution to journalArticle

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Abstract

Islet transplantation is limited by islet graft failure due to poor revascularization, host immune rejection and nonspecific inflammatory response. Delivery of human vascular endothelial growth factor (hVEGF) gene to the islets is likely to promote islet revascularization and survival. We used a bicistronic adenoviral vector encoding hVEGF and CpG-free allele of green fluorescent protein (Adv-GFP-hVEGF) and introduced into human pancreatic islets by transfection. We found that transfection efficiency and apoptosis were dependent on the multiplicity of infection (MOI). Compared to Adv-GFP transfected and nontransfected islets, the levels of hVEGF secreted from Adv-GFP-hVEGF transfected islets were higher and exhibit a linear relationship between hVEGF expression and MOI (10-5000). Persistent, but low level expression of hVEGF from nontransfected islets was also observed. This may be due to expression of the endogenous hVEGF gene under hypoxic conditions. The levels of DNA fragmentation determined by ELISA of islet lysates were dependent on the MOI of Adv-GFP-hVEGF. On glucose challenge, insulin release from transfected islets was comparable to nontransfected islets. Immunohistochemical staining for hVEGF was very high in Adv-GFP-hVEGF transfected islets. Weak staining was also observed for hCD31 in both transfected and nontransfected islets. These findings suggest that Adv-GFP-hVEGF is a potential candidate for promoting islet revascularization.

Original languageEnglish (US)
Pages (from-to)1105-1116
Number of pages12
JournalGene Therapy
Volume11
Issue number14
DOIs
StatePublished - Jul 1 2004

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Islets of Langerhans
Adenoviridae
Vascular Endothelial Growth Factor A
Genes
Transfection
human VEGFA protein
Infection
Staining and Labeling
Islets of Langerhans Transplantation
DNA Fragmentation
Green Fluorescent Proteins
Enzyme-Linked Immunosorbent Assay
Alleles
Insulin
Apoptosis
Transplants
Glucose

All Science Journal Classification (ASJC) codes

  • Molecular Medicine
  • Molecular Biology
  • Genetics

Cite this

Cheng, K., Fraga, D., Zhang, C., Kotb, M., Gaber, A. O., Guntaka, R., & Mahato, R. I. (2004). Adenovirus-based vascular endothelial growth factor gene delivery to human pancreatic islets. Gene Therapy, 11(14), 1105-1116. https://doi.org/10.1038/sj.gt.3302267

Adenovirus-based vascular endothelial growth factor gene delivery to human pancreatic islets. / Cheng, K.; Fraga, D.; Zhang, C.; Kotb, M.; Gaber, A. O.; Guntaka, Ramareddy; Mahato, R. I.

In: Gene Therapy, Vol. 11, No. 14, 01.07.2004, p. 1105-1116.

Research output: Contribution to journalArticle

Cheng, K, Fraga, D, Zhang, C, Kotb, M, Gaber, AO, Guntaka, R & Mahato, RI 2004, 'Adenovirus-based vascular endothelial growth factor gene delivery to human pancreatic islets', Gene Therapy, vol. 11, no. 14, pp. 1105-1116. https://doi.org/10.1038/sj.gt.3302267
Cheng, K. ; Fraga, D. ; Zhang, C. ; Kotb, M. ; Gaber, A. O. ; Guntaka, Ramareddy ; Mahato, R. I. / Adenovirus-based vascular endothelial growth factor gene delivery to human pancreatic islets. In: Gene Therapy. 2004 ; Vol. 11, No. 14. pp. 1105-1116.
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