combination of vascular endothelial growth factor (VEGF) and thymidine phosphorylase (TP) to improve angiogenic gene therapy.
To improve current angiogenic gene therapy with a vascular endothelial growth factor (VEGF)-encoding plasmid (Baumgartner et al. Circulation 1998; 97: 1114-23 [1]; Kusumanto et al. Fifth Annual Meeting of the American Society of Gene Therapy, Boston, 2002, Abstr. 621 [2]), we have generated a combination plasmid, encoding the VEGF gene and the thymidine phosphorylase (TP, also known as platelet-derived endothelial growth factor (PD-ECGF) or gliostatin (GLS)) gene: phVEGF165-TP.MB. Upon transfection in COS-7 cells both gene products were expressed and functional as shown by Western blots, ELISAs and bioassays. Culture supernatants of COS-7 cells transfected with this plasmid were able to induce endothelial proliferation. In an in vitro angiogenesis assay with recombinant proteins, TP was able to increase VEGF-induced tube formation. The phVEGF165-TP.MB plasmid is therefore a promising candidate for in vivo angiogenesis studies.
Bouïs
Diane
D
Department of Medical Oncology, University Hospital Groningen, The Netherlands.
Boelens
Mirjam C
MC
Peters
Erna
E
Koolwijk
Pieter
P
Stob
Gerrie
G
Kema
Ido P
IP
Klinkenberg
Marco
M
Mulder
Nanno H
NH
Hospers
Geke A P
GA
D000818:Animals; D019556:COS Cells; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D017077:Culture Media, Conditioned / Q000494:pharmacology; D004359:Drug Therapy, Combination; D004730:Endothelium, Vascular / Q000166:cytology; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D006801:Humans; D018919:Neovascularization, Physiologic / Q000187:drug effects*; D013939:Thymidine Phosphorylase / Q000008:administration & dosage* / Q000235:genetics / Q000494:pharmacology; D013941:Thymine / Q000096:biosynthesis; D014162:Transfection; D014471:Umbilical Veins; D042461:Vascular Endothelial Growth Factor A / Q000008:administration & dosage* / Q000235:genetics / Q000494:pharmacology
D017077:Culture Media, Conditioned; D042461:Vascular Endothelial Growth Factor A; D013939:Thymidine Phosphorylase; D013941:Thymine