Single Intraluminal Delivery of Antisense cdc2 Kinase and Proliferating- Cell Nuclear Antigen Oligonucleotides Results in Chronic Inhibition of Neointimal Hyperplasia

• Published in: Proceedings of the National Academy of Sciences - PNAS Vol. 90; no. 18; pp. 8474 - 8478
• Main Authors: Morishita, Ryuichi, Gibbons, Gary H., Ellison, Kristin E., Nakajima, Masatoshi, Zhang, Lunan, Kaneda, Yasufumi, Ogihara, Toshio, Dzau, Victor J.
• Format: Journal Article
• Language: English
• Published: Washington, DC National Academy of Sciences of the United States of America 15.09.1993; National Acad Sciences; National Academy of Sciences
• Subjects: Angioplasty; Animals; Aorta - cytology; Aorta - drug effects; Base Sequence; Biological and medical sciences; Cardiology. Vascular system; Carotid Arteries - drug effects; Carotid Arteries - pathology; Carotid Artery Injuries; CDC2 Protein Kinase - genetics; Cell Cycle - drug effects; Cell cycle proteins; Cell growth; Cells, Cultured; Cellular biology; Coronary heart disease; DNA; DNA Replication - drug effects; Drug Carriers; Genes; Heart; Hyperplasia; Lesions; Liposomes; Male; Medical sciences; Messenger RNA; Molecular Sequence Data; Muscle, Smooth, Vascular - cytology; Muscle, Smooth, Vascular - drug effects; Muscle, Smooth, Vascular - injuries; Neointima; Nuclear Proteins - genetics; Oligodeoxyribonucleotides - pharmacology; Oligonucleotides, Antisense - administration & dosage; Oligonucleotides, Antisense - metabolism; Oligonucleotides, Antisense - pharmacology; Physical trauma; Polymerase Chain Reaction; Proliferating Cell Nuclear Antigen; Proteins; Rats; Rats, Sprague-Dawley; RNA; Sendai virus; Transfection; Delivery system; Rat; Enzyme; Rodentia; DNA binding protein; Instrumentation therapy; Cardiovascular disease; Instrumental dilatation; Coronary heart disease; In vitro; Intraluminal; Restenosis; In vivo; Vertebrata; Target; Mammalia; Animal; Blood vessel; Targeted drug; Antisense DNA; Inhibition; Gene therapy; Nuclear protein
• ISSN: 0027-8424; 1091-6490
• DOI: 10.1073/pnas.90.18.8474

To develop an effective strategy to prevent neointima formation after angioplasty injury, we have identified cell-cycle regulatory proteins as targets for inhibition by using antisense oligonucleotides (ODNs). We utilized an intraluminal molecular delivery method that employs the protein coat of a Sendai virus complexed with liposomes that enhances markedly the efficiency of ODNs uptake. First, we examined the effect of antisense cdc2 kinase and proliferating-cell nuclear antigen (PCNA) ODNs in vitro. Cotransfection of antisense cdc2 kinase and PCNA ODNs inhibited serum-stimulated vascular smooth muscle cell growth, whereas antisense cdc2 kinase ODNs alone or PCNA ODNs alone failed to show any inhibitory effect. Transfection of the combination of antisense cdc2 kinase and PCNA ODNs into balloon-injured arteries in vivo provided a marked decrease in cdc2 and PCNA mRNA expression as determined by reverse transcription-PCR, compared to sense controls. Antisense ODN treatment significantly inhibited the increase in DNA synthesis induced by balloon injury. Moreover, antisense ODN administration inhibited completely neointima formation at 2 weeks after angioplasty in an apparent dose-dependent manner. Moreover, the inhibitory effect of antisense ODN on neointima formation persisted up to 8 weeks after a single transfection. The present study documents that a single intraluminal molecular delivery of combined cdc2 kinase and PCNA antisense ODNs results in a sustained inhibition of neointima formation in the rat carotid balloon-injury model.