December 2002
Volume 43, Issue 13
ARVO Annual Meeting Abstract  |   December 2002
Constrction of a Lentiviral System Harboring the Soluble Vegf Receptor 2 (KDR), a New Angiostatic Reagent
Author Affiliations & Notes
  • RC Murthy
    Retina Casey Eye Institute Portland OR
  • Footnotes
    Commercial Relationships   R.C. Murthy, None.
Investigative Ophthalmology & Visual Science December 2002, Vol.43, 1761. doi:
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      RC Murthy; Constrction of a Lentiviral System Harboring the Soluble Vegf Receptor 2 (KDR), a New Angiostatic Reagent . Invest. Ophthalmol. Vis. Sci. 2002;43(13):1761.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract: : Purpose: Vascular endothelial factor (VEGF) is a specific and potent mitogen for vascular endothelial cells, via binding and activation of the membrane bound vascular endothelial growth receptors, FMS-tyrosine related kinase 1 (FLT1/VEGFR1) and kinase insert domain receptor (KDR/VEGFR2). Soluble (non-membrane bound) form of the VEGR2 competitively inhibits the binding and activation of the membrane bound receptors. We wish to develop a reagent that can effectively deliver sVEGR2 to areas of angiogenesis within the eye. Methods: KDR cDNA was amplified using a BamHI forward linker primer and an EcoRI reverse linker primer. The amplified product was cloned into the lentiviral vector pHR'-IRES-eGFP under the control of the CMV promoter. The insert was sequenced and replication-defective sKDR lentivirus was prepared by co-transfection of three plasmids, pHR'CMV-sKDR-IRES-eGFP, pCMV Δ;R8.91 and pMD.G, into 293T cells. A 4 x 4mm nylon mesh soaked with the recombinant KDR lentivirus was inserted into a corneal stromal pocket in one eye of New Zealand White rabbits, the contralateral eye served as a non-transduced control. Results: A 2274 bp cDNA coding for 757 aa soluble form of the human VEGF receptor 2, was successfully cloned into a pHR'-CMV-IRES-eGFP plasmid. Replication deficient lentiviral particles containing the soluble form of VEGFR2 have been produced. Conclusion: VEGF is a primary inducer of angiogenesis, via activation of a membrane bound receptor KDR/VEGR2. Competitive inhibition with the soluble form of KDR should reduce the neovascular response. Development of an efficient lentiviral sKDR delivery system is a useful tool to study and treat neovascular disease. Supported by the Clayton Foundation for Research and Research to Prevent Blindness. None

Keywords: 419 gene transfer/gene therapy • 483 neovascularization 

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