Efficient adenoviral gene transfer to kidney cortical vasculature utilizing a fiber modified vector
✍ Scribed by Glenn A. McDonald; Guoming Zhu; Yongsheng Li; Imre Kovesdi; Thomas J. Wickham; Vikas P. Sukhatme
- Publisher
- John Wiley and Sons
- Year
- 1999
- Tongue
- English
- Weight
- 633 KB
- Volume
- 1
- Category
- Article
- ISSN
- 1099-498X
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✦ Synopsis
Background The a v integrin is present in vascular endothelium, including that of the kidney. It is also upregulated in the presence of in¯ammatory cytokines and in some neoplasms. In an effort to transduce vascular endothelial cells, we compare in vitro and in vivo adenoviral gene transfer of a vector with a high af®nity peptide ligand to the a v integrin incorporated into the ®ber coat protein AdZ.F(RGD) to an unmodi®ed vector, AdZ.
Methods Cell transduction assays were performed on human umbilical vein endothelial cells (HuVEC) and human pulmonary epithelial cells (A549). In vitro competition assays were performed in the presence of either wild type (F5K) or chimeric (F5K(RGD)) soluble recombinant ®ber protein. Transduction ef®ciency was determined by quantitative b-galactosidase activity. In vivo gene transfer was compared infusing either AdZ or AdZ.F(RGD) into the left renal artery of the rat and assaying b-galactosidase staining of the kidney. Gene transfer was also evaluated in the presence of a competitive RGD or control RGE peptide.
Results
There was a marked increase in transgene expression in HuVEC cells with AdZ.F(RGD) as compared to AdZ. The increased expression with AdZ.F(RGD) was more prominent in the endothelial as opposed to the epithelial cell line. Furthermore pre-incubation of these cells with either F5K or F5K(RGD) soluble ®ber protein markedly decreased b-galactosidase activity of AdZ, whereas only the F5K(RGD) decreased b-galactosidase activity of AdZ.F(RGD). AdZ.F(RGD) also resulted in signi®cantly enhanced b-galactosidase expression in the vascular endothelium of the kidney (for comparable amounts of virus injected) as well as signi®cantly higher gene transfer to cortical vasculature. Coinfusion of an RGD peptide with AdZ.F(RGD) blocked gene transfer whereas a control RGE peptide did not.
Conclusion
We conclude that incorporating a high af®nity peptide ligand into the adenoviral ®ber protein can preferentially enhance in vitro and in vivo adenoviral transfection ef®ciency in endothelial cells. Enhancing transfection ef®ciency will not only broaden the scope of disease processes addressed with adenoviral vectors but also allow the use of lower titer, thereby limiting toxicity. This vector has additional potential to transduce endothelial cells within tumors or ischemic tissue where a v integrins are upregulated.