Inhibition of angiogenesis in vitro by alphav integrin-directed antisense oligonucleotides.
The integrin alpha v beta 3 plays a central role in angiogenesis. In this study, we used antisense oligodeoxyribonucleotides (ONs) directed against the alpha v subunit of alpha v beta 3 to inhibit integrin expression. Ten ON sequences, which were selected by systematic alignment of computer-predicted secondary structures of alpha v mRNA, were transfected into human umbilical vein endothelial cells (HUVECs). Following stimulation by PMA, five antisense ONs significantly inhibited alpha v mRNA and protein expression in activated HUVEC at a concentration of 0.05 mciroM with complete prevention of PMA-induced alpha v up-regulation by the most potent antisense ON. Inhibition of alpha v expression was associated with significant inhibition of migration of HUVEC by 28% and had no effect on proliferation and apoptosis. Moreover, transfection of antisense ON inhibited the formation of tube-like structures of HUVEC in Matrigel by 44%. In a cell culture model of angiogenesis consisting of a co-culture of endothelial cells with fibroblasts, transfection of antisense ONs resulted in an inhibition of tube formation of 61%. In conclusion, alpha v antisense ONs are potent inhibitors of angiogenesis in vitro. They might, therefore, be a therapeutic alternative to antagonists, which directly bind to alpha v integrins, and might be useful for the treatment of malignant tumors and hematological malignancies.
Kronenwett
Ralf
R
Department of Hematology, Oncology and Clinical Immunology, University of Düsseldorf, Düsseldorf, Germany. kronenwett@med.uni-duesseldorf.de
Gräf
Thorsten
T
Nedbal
Wolfgang
W
Weber
Markus
M
Steidl
Ulrich
U
Rohr
Ulrich-Peter
UP
Möhler
Thomas
T
Haas
Rainer
R
D017209:Apoptosis; D001483:Base Sequence; D002455:Cell Division; D002465:Cell Movement; D002478:Cells, Cultured; D003094:Collagen / Q000494:pharmacology; D015536:Down-Regulation; D004338:Drug Combinations; D004730:Endothelium, Vascular / Q000166:cytology; D005434:Flow Cytometry; D006801:Humans; D039564:Integrin alphaV / Q000235:genetics*; D007797:Laminin / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D008969:Molecular Sequence Data; D009389:Neovascularization, Pathologic; D009690:Nucleic Acid Conformation; D016376:Oligonucleotides, Antisense / Q000494:pharmacology*; D011509:Proteoglycans / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014162:Transfection; D014471:Umbilical Veins / Q000166:cytology; D015854:Up-Regulation
D004338:Drug Combinations; D039564:Integrin alphaV; D007797:Laminin; D016376:Oligonucleotides, Antisense; D011509:Proteoglycans; D012333:RNA, Messenger; C059485:matrigel; D003094:Collagen