Sequence-specific antiproliferative effects of antisense and end-capping-modified antisense oligodeoxynucleotides targeted against the 5'-terminus of basic-fibroblast-growth-factor mRNA in coronary smooth muscle cells.
Basic fibroblast growth factor (bFGF), a potent mitogen for arterial smooth muscle cells, has been shown to play a fundamental role in the pathogenesis of arteriosclerosis and restenosis by stimulating the proliferation of vascular smooth muscle cells. We found that partially phosphorothioate-modified 15-residue antisense oligodeoxynucleotides complementary to bFGF mRNA at 0.1-2.0 microM block growth and division of cultured human and bovine coronary smooth muscle cells in a dose-dependent manner. The effect is sequence specific at low (0.1-0.5 microM) nontoxic concentrations. It is associated with inhibition of expression of pericellular and intracellular bFGF, with a decreased de novo synthesis of bFGF and is partly reversible by the addition of exogenous (recombinant) bFGF. The antisense effect lasts 48-72 h and diminishes thereafter. If the antisense oligodeoxynucleotide medium is replaced by an oligonucleotide-free medium after 24 h, the [3H]thymidine incorporation rate returns to control levels. Under the same conditions, the corresponding sense oligodeoxynucleotide exerts negligible nonspecific inhibitory actions. The antiproliferative potency of the 15-residue antisense oligodeoxynucleotide is markedly enhanced by adding 3-4 nonbase-pairing guanosine residues at the 5'- and 3'-termini of the 15-residue antisense oligonucleotide. The data implicate bFGF in the process of smooth muscle cell proliferation and an effective and specific antiproliferative potency of bFGF-specific antisense oligonucleotides. The results point to possible new therapeutic strategies for the use of antisense methodology in the suppression of post-angioplasty restenosis.
Schmidt
A
A
Institute for Arteriosclerosis Research, Division of Molecular Cardiology, University of Münster, Germany. annschm@uni-muenster.de
Sindermann
J
J
Peyman
A
A
Uhlmann
E
E
Will
D W
DW
Müller
J G
JG
Breithardt
G
G
Buddecke
E
E
D000818:Animals; D002417:Cattle; D002455:Cell Division / Q000187:drug effects; D003331:Coronary Vessels / Q000187:drug effects*; D016222:Fibroblast Growth Factor 2 / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D006151:Guanosine / Q000737:chemistry; D006801:Humans; D009131:Muscle, Smooth, Vascular / Q000166:cytology* / Q000187:drug effects; D016376:Oligonucleotides, Antisense / Q000737:chemistry* / Q000494:pharmacology*; D011743:Pyrimidines / Q000737:chemistry; D012333:RNA, Messenger / Q000187:drug effects; D015398:Signal Transduction; D013329:Structure-Activity Relationship
D016376:Oligonucleotides, Antisense; D011743:Pyrimidines; D012333:RNA, Messenger; D016222:Fibroblast Growth Factor 2; D006151:Guanosine; C030986:pyrimidine