IN-VIVO ANGIOGENESIS INDUCED BY RECOMBINANT ADENOVIRUS VECTORS CODINGEITHER FOR SECRETED OR NONSECRETED FORMS OF ACIDIC FIBROBLAST GROWTH-FACTOR

In vivo gene transfer of angiogenic growth factors represents a potent ial approach to the treatment of ischemic diseases, The present study examined the in vitro and in vivo effects of two replication-deficient recombinant adenovirus (Ad) vectors coding for human acidic fibroblas t growth factor (aFGF(1-154)). One vector codes for the nonsecreted fo rm of the peptide (AdCMV.aFGF(1-154)), and the other vector codes for a recombinant, secreted form (AdCMV.sp+aFGF(1-154)). AdCMV.NLS beta ga l, an adenovirus vector coding for beta-galactosidase (beta-Gal), was used as a control, Assessment of proliferation of starved human umbili cal vein endothelial cells infected with AdCMV.aFGF(1-154) and AdCMV.s p+aFGF(1-154) (20 pfu/cell) showed approximately 6- and 10-fold increa se in cell number over control, respectively, Infection with AdCMV.spaFGF(1-154) and with AdCMV.aFGF(1-154) enhanced endothelial cell diffe rentiation into capillary-like structures in vitro, However, this effe ct was significantly more pronounced with AdCMV.sp+aFGF(1-154) than wi th AdCMV.aFGF(1-154). Angiogenesis in vivo was assessed by injecting s ubcutaneously into mice 750 mu l of reconstituted basement membrane pr oteins (Matrigel) and the Ad vectors (2 x 10(8) pfu), After 14 days, t here was histologic evidence of neovascularization in the animal's tis sue surrounding the Matrigel plugs with AdCMV.aFGF(1-154) and AdCMV.sp +aFGF(1-154). Further, the hemoglobin content of the Matrigel plugs wi th AdCMV.aFGF(1-154) and with AdCMV.sp+aFGF(1-154) was, respectively, 2.3- and 2.6-fold higher than with AdCMV.NLS beta gal. Together, these observations support the concept that adenovirus vectors coding for v arious forms of acidic FGF(1-154) may be used to induce angiogenesis i n vivo and may provide a new therapeutic approach to ischemic diseases .