title: Inhibition of intimal hyperplasia after stenting by over-expression of p15: a member of the INK4 family of cyclin-dependent kinase inhibitors.
abstract: We evaluated the role of p15(Ink4), a member of the INK4 family of CDK inhibitors on vascular smooth muscle cells (VSMCs) proliferation, cell cycle progression and intimal hyperplasia after stenting. Aortic VSMCs transduced with either adenovirus encoding for p15(Ink4) or β-galactosidase were assessed for DNA synthesis, cell cycle progression, and pRb phosphorylation. Rabbit carotid arteries were stented and treated with peri-adventitial delivery of saline or adenovirus encoding for p15(Ink4) or β-galactosidase. p15(Ink4) transgene and protein expression were evaluated at 24 h and 72 h, respectively. In-stent cell proliferation was evaluated by BrdU at day 7. Histomorphometric analysis of in-stent intimal hyperplasia was performed at 10 weeks. Human p15(Ink4) DNA was detected in transduced VSMCs at 24h. p15(Ink4) over-expression reduced VSMCs DNA synthesis by 60%. Cell cycle progression was inhibited, with a 30% increase in G1 population accompanied by inhibition of pRb phosphorylation. Human p15(Ink4) transgene was identified in transduced stented arteries but not in control arteries. p15(Ink4) immunostaining was increased and cell proliferation significantly reduced by 50% in p15(Ink4) transduced arteries. Intimal cross-sectional area (CSA) of p15(Ink4)-treated group was significantly lower than the β-gal treated and non-transduced groups (p=0.008). There were no differences in the intimal or medial inflammatory response between groups. p15(Ink4) over-expression blocks cell cycle progression leading to inhibition of VSMCs proliferation. Peri-adventitial delivery of p15(Ink4) significantly inhibits in-stent intimal hyperplasia.
authors.affiliation: Interventional Cardiology Unit, Chaim Sheba Medical Center, Tel Hashomer, Sackler School of Medicine, Tel-Aviv University, Tel-Aviv, Israel.
authors.forename: Amit
authors.initials: A
authors.lastname: Segev
authors.forename: Nafiseh
authors.initials: N
authors.lastname: Nili
authors.forename: Beiping
authors.initials: B
authors.lastname: Qiang
authors.forename: Azriel B
authors.initials: AB
authors.lastname: Osherov
authors.forename: Frank J
authors.initials: FJ
authors.lastname: Giordano
authors.forename: Ronen
authors.initials: R
authors.lastname: Jaffe
authors.forename: Jack
authors.initials: J
authors.lastname: Gauldie
authors.forename: John D
authors.initials: JD
authors.lastname: Sparkes
authors.forename: Ashley R
authors.initials: AR
authors.lastname: Fraser
authors.forename: Michelle
authors.initials: M
authors.lastname: Ladouceur-Wodzak
authors.forename: Jagdish
authors.initials: J
authors.lastname: Butany
authors.forename: Bradley H
authors.initials: BH
authors.lastname: Strauss
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D002339:Carotid Arteries / Q000378:metabolism / Q000473:pathology; D048708:Cell Growth Processes / Q000502:physiology; D002478:Cells, Cultured; D050762:Cyclin-Dependent Kinase Inhibitor p15 / Q000096:biosynthesis / Q000235:genetics* / Q000378:metabolism*; D016193:G1 Phase / Q000235:genetics; D015316:Genetic Therapy / Q000379:methods; D006801:Humans; D006965:Hyperplasia / Q000209:etiology / Q000378:metabolism / Q000473:pathology; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D010766:Phosphorylation; D011817:Rabbits; D051381:Rats; D016160:Retinoblastoma Protein / Q000378:metabolism; D015607:Stents / Q000009:adverse effects*; D014161:Transduction, Genetic; D019076:Transgenes; D017539:Tunica Intima / Q000378:metabolism / Q000473:pathology*; D001616:beta-Galactosidase / Q000096:biosynthesis / Q000235:genetics / Q000378:metabolism
chemical_list: D050762:Cyclin-Dependent Kinase Inhibitor p15; D016160:Retinoblastoma Protein; D001616:beta-Galactosidase