The antiproliferative activity of c-myb and c-myc antisense oligonucleotides in smooth muscle cells is caused by a nonantisense mechanism.
Smooth muscle cell (SMC) proliferation is thought to play a major role in vascular restenosis after angioplasty and is a serious complication of the procedure. Developing antisense (AS) oligonucleotides as therapeutics is attractive because of the potentially high specificity of binding to their targets, and several investigators have reported inhibition of SMC proliferation in vitro and in vivo by using AS strategies. We report here the results of our experiments on vascular SMCs using AS oligonucleotides directed toward c-myb and c-myc. We found that significant inhibition of SMC proliferation occurred with these specific AS sequences but that this inhibition was clearly not via a hybridization-dependent AS mechanism. Rather, inhibition was due to the presence of four contiguous guanosine residues in the oligonucleotide sequence. This was demonstrated in vitro in primary cultures of SMCs and in arteries ex vivo. The ex vivo model developed here provides a rapid and effective system in which to screen potential oligonucleotide drugs for restenosis. We have further explored the sequence requirements of this non-AS effect and determined that phosphorothioate oligonucleotides containing at least two sets of three or four consecutive guanosine residues inhibit SMC proliferation in vitro and ex vivo. These results suggest that previous AS data obtained using these and similar, contiguous guanosine-containing AS sequences be reevaluated and that there may be an additional class of nucleic acid compounds that have potential as antirestenosis therapeutics.
Burgess
T L
TL
Department of Mammalian Cell Molecular Biology, Amgen Inc., Amgen Center, Thousand Oaks, CA 91320-1789, USA.
Fisher
E F
EF
Ross
S L
SL
Bready
J V
JV
Qian
Y X
YX
Bayewitch
L A
LA
Cohen
A M
AM
Herrera
C J
CJ
Hu
S S
SS
Kramer
T B
TB
D000818:Animals; D001011:Aorta / Q000166:cytology / Q000187:drug effects; D001483:Base Sequence; D001973:Bromodeoxyuridine; D002453:Cell Cycle; D002455:Cell Division / Q000187:drug effects*; D002478:Cells, Cultured; D003062:Codon; D016259:Genes, myc; D007700:Kinetics; D008969:Molecular Sequence Data; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000187:drug effects*; D016376:Oligonucleotides, Antisense / Q000138:chemical synthesis / Q000494:pharmacology*; D009857:Oncogenes; D011518:Proto-Oncogene Proteins / Q000235:genetics; D020598:Proto-Oncogene Proteins c-myb; D011817:Rabbits; D013329:Structure-Activity Relationship
D003062:Codon; D016376:Oligonucleotides, Antisense; D011518:Proto-Oncogene Proteins; D020598:Proto-Oncogene Proteins c-myb; D001973:Bromodeoxyuridine