Heparin-independent mitogenicity in an endothelial and smooth muscle cell chimeric growth factor (S130K-HBGAM).
BACKGROUND
Through site-directed mutagenesis we have created a favorable fibroblast growth factor-1 (FGF-1) mutant (S130K) and linked it to a heparin-binding growth-associated molecule (HBGAM) to form the chimera S130K-HBGAM creating a heparin-independent, endothelial cell (EC)-specific mitogen.


METHODS
The proliferative responses of primary canine carotid artery smooth muscle cells (SMC) and jugular vein EC to FGF-1, S130K, or S130K-HBGAM, with and without heparin (5 U/mL), was quantitated by measuring tritiated thymidine uptake over 24 hours and expressing the results as percent of positive control (20% fetal bovine serum [FBS]) for group comparison.


RESULTS
Unlike FGF-1, both S130K and S130K-HBGAM are heparin-independent mitogens for EC and SMC. S130K-HBGAM was equivalent to FGF-1 with heparin at 6 nmol/L. S130K-HBGAM did not demonstrate relative EC specificity in this assay.


CONCLUSIONS
At higher concentrations, S130K-HBGAM is a potent, heparin-independent EC and SMC mitogen. Co-culture assays and in vivo delivery models may demonstrate EC specificity not identified in this single cell type proliferation assay.
Brewster
Luke P
LP
Department of Surgery, Loyola University Medical Center, 2160 South First Ave., Maywood, IL 60153, USA.
Brey
Eric M
EM
Tassiopoulos
Apostolos K
AK
Xue
Lian
L
Maddox
Ewa
E
Armistead
David
D
Burgess
Wilson H
WH
Greisler
Howard P
HP
D000704:Analysis of Variance; D000818:Animals; D001483:Base Sequence; D002339:Carotid Arteries / Q000166:cytology; D049109:Cell Proliferation / Q000187:drug effects; D002478:Cells, Cultured; D004285:Dogs; D004730:Endothelium, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D016220:Fibroblast Growth Factor 1 / Q000235:genetics / Q000378:metabolism* / Q000494:pharmacology*; D006493:Heparin / Q000494:pharmacology*; D008934:Mitogens / Q000494:pharmacology*; D023421:Models, Animal; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects / Q000378:metabolism*; D032389:Myocytes, Smooth Muscle / Q000187:drug effects / Q000378:metabolism*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011336:Probability; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D011994:Recombinant Proteins; D012680:Sensitivity and Specificity
D008934:Mitogens; D011993:Recombinant Fusion Proteins; D011994:Recombinant Proteins; C494171:S130K-HBGAM chimeric protein; C422626:fibroblast growth factor-1, S130K substituted; D016220:Fibroblast Growth Factor 1; D006493:Heparin