Objective: Decoy transfection is a significant methodology for suppressing gene activation. The decoy can be synthesized easily; however, the major obstacle is the difficulty involved in effective in vivo delivery. Methods and Results: We used a fluorescein-labeled decoy to investigate the ultrasound-microbubble-mediated in vivo delivery in normal and injured mouse arteries. We showed that this approach could deliver the decoy into target tissues. In addition, we performed in vivo NF-ĸB decoy transfection into murine injured arteries using the ultrasound-microbubble method. Murine femoral arteries were injured using flexible wires to establish arterial injury. Pathologically, neointima/media areas in the NF-ĸB decoy transfection using ultrasound-microbubble group showed less than those in the control groups. Immunohistochemistry revealed that enhanced expression of inflammatory factors was observed in nontreated injured arteries, while the NF-ĸB decoy suppressed the expression. Conclusion: We revealed that ultrasound-microbubble delivery of the decoy is effective for transfection into target organs. We also indicated that NF-ĸB decoy transfection using this method has potential for the suppression of neointimal formation. Ultrasound-mediated transfection of the decoy can be beneficial for the clinical treatment of restenosis after coronary intervention and other cardiovascular diseases.

Keywords:
Ultrasound, Decoy, Gene therapy, Smooth muscle cells, Arteriosclerosis
1.
Leimgruber PP, Roubin GS, Hollman J, Cotsonis GA, Meier B, Douglas JS, King SB Jr, Gruentzig AR: Restenosis after successful coronary angioplasty in patients with single-vessel disease. Circulation 1986;73:710–717.
2.
Haude M, Konorza TF, Kalnins U, Erglis A, Saunamaki K, Glogar HD, Grube E, Gil R, Serra A, Richardt HG, Sick P, Erbel R; Heparin-COAted STents in small coronary arteries Trial Investigators: Heparin-coated stent placement for the treatment of stenoses in small coronary arteries of symptomatic patients. Circulation 2003;107:1265–1270.
3.
Holmes DR Jr: In-stent restenosis. Rev Cardiovasc Med 2001;2:115–119.
4.
Squadrito F, Deodato B, Bova A, Marini H, Saporito F, Calo M, Giacca M, Minutoli L, Venuti FS, Caputi AP, Altavilla D: Crucial role of nuclear factor-kappaB in neointimal hyperplasia of the mouse carotid artery after interruption of blood flow. Atherosclerosis 2003;166:233–242.
5.
Suzuki J, Morishita R, Amano J, Kaneda Y, Isobe M: Decoy against nuclear factor-kappa B attenuates myocardial cell infiltration and arterial neointimal formation in murine cardiac allografts. Gene Ther 2000;7:1847–1852.
6.
Suzuki J, Ito H, Gotoh R, Morishita R, Egashira K, Isobe M: Initial clinical cases using an NF-κB decoy at the site of the coronary stenting for prevention of restenosis. Circ J 2004;68:270–271.
7.
Miller DL, Pislaru SV, Greenleaf JE: Sonoporation: mechanical DNA delivery by ultrasonic cavitation. Somat Cell Mol Genet 2002;27:115–134.
8.
Kim HJ, Greenleaf JF, Kinnick RR, Bronk JT, Bolander ME: Ultrasound-mediated transfection of mammalian cells. Hum Gene Ther 1996;7:1339–1346.
9.
Yamashita Y, Shimada M, Tachibana K, Harimoto N, Tsujita E, Shirabe K, Miyazaki J, Sugimachi K: In vivo gene transfer into muscle via electro-sonoporation. Hum Gene Ther 2002;13:2079–2084.
10.
Azuma H, Tomita N, Kaneda Y, Koike H, Ogihara T, Katsuoka Y, Morishita R: Transfection of NFkappaB-decoy oligodeoxynucleotides using efficient ultrasound-mediated gene transfer into donor kidneys prolonged survival of rat renal allografts. Gene Ther 2003;10:415–425.
11.
Nakashima M, Tachibana K, Iohara K, Ito M, Ishikawa M, Akamine A: Induction of reparative dentin formation by ultrasound-mediated gene delivery of growth/differentiation factor 11. Hum Gene Ther 2003;14:591–597.
12.
Taniyama Y, Tachibana K, Hiraoka K, Namba T, Yamasaki K, Hashiya N, Aoki M, Ogihara T, Yasufumi K, Morishita R: Local delivery of plasmid DNA into rat carotid artery using ultrasound. Circulation 2002;105:1233–1239.
13.
Yoshimura S, Morishita R, Hayashi K, Yamamoto K, Nakagami H, Kaneda Y, Sakai N, Ogihara T: Inhibition of intimal hyperplasia after balloon injury in rat carotid artery model using cis-element ‘decoy’ of nuclear factor-kappaB binding site as a novel molecular strategy. Gene Ther 2001;8:1635–1642.
14.
Yamasaki K, Asai T, Shimizu M, Aoki M, Hashiya N, Sakonjo H, Makino H, Kaneda Y, Ogihara T, Morishita R: Inhibition of NFkappaB activation using cis-element ‘decoy’ of NFkappaB binding site reduces neointimal formation in porcine balloon-injured coronary artery model. Gene Ther 2003;10:356–364.
15.
Lindner V, Fingerle J, Reidy MA: Mouse model of arterial injury. Circ Res 1993;73:792–796.
16.
Nakashima H, Aoki M, Miyake T, Kawasaki T, Iwai M, Jo N, Oishi M, Kataoka K, Ohgi S, Ogihara T, Kaneda Y, Morishita R: Inhibition of experimental abdominal aortic aneurysm in the rat by use of decoy oligodeoxynucleotides suppressing activity of nuclear factor kappaB and ets transcription factors. Circulation 2004;109:132–138.
17.
Suzuki J, Isobe M, Morishita R, Aoki M, Horie S, Okubo Y, Kaneda Y, Sawa Y, Matsuda H, Ogihara T, Sekiguchi M: Prevention of graft coronary arteriosclerosis by antisense cdk2 kinase oligonucleotide. Nat Med 1997;3:900–903.
18.
Futamatsu H, Suzuki J, Kosuge H, Yokoseki O, Kamada M, Ito H, Inobe M, Isobe M, Uede T: Attenuation of experimental autoimmune myocarditis by blocking activated T cells through inducible costimulatory molecule pathway. Cardiovasc Res 2003;59:95–104.
19.
Morishita R, Tomita N, Kaneda Y, Ogihara T: Molecular therapy to inhibit NFkappaB activation by transcription factor decoy oligonucleotides. Curr Opin Pharmacol 2004;4:139–146.
20.
Monaco C, Paleolog E: Nuclear factor kappaB: a potential therapeutic target in atherosclerosis and thrombosis. Cardiovasc Res 2004;61:671–682.
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2005
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