title: Polymer-based delivering of shRNA to rabbit aortic smooth muscle cells suppressed the expression of IGF-1R in vitro and in vivo.
abstract: Restenosis is one of clinical limitations for vein graft in coronary bypass graft. It has been proved that signal pathway IGF-1 and its receptor (IGF-1R) activated by hemodynamic mechanical stretch are responsible for the vascular smooth muscle cells proliferation in vein graft neointima formation. Unfortunately, there is no routinely successful method to resolve this problem. Gene delivering to vein graft possesses great therapeutic potential to prevent neointima formation. Polymer is one kind of nanoparticles, which can activate the process of endocytosis of cells. In this study, we evaluated the transfection efficiency and therapeutic potential of polymer-based transfection of plasmids expressing GFP and shRNAs targeting IGF-1R (pGFPshIGF-1Rs) to smooth muscle cells and rabbit external jugular vein graft. Results showed that polymer-based transfection provided high efficiency of transgene expression in smooth muscle cells in vitro. In vitro, IGF-1R-specific shRNA transfected by polymer inhibited IGF-1R protein expression by 52 ± 3.6%, when compared with mock transfected cells. In vivo delivering efficiency of pGFPshIGF-1R plasmid into the rabbit external jugular vein graft was significantly high in the polymer-based transfection group, when compared with negative control group. In vivo, polymer-based transfection IGF-1R-specific shRNA efficiently inhibited the expression of IGF-1R protein by 77 ± 3.6%, 65.6 ± 4.9%, and 76.7 ± 4.3% at 24, 48, and 72 h, respectively, when compared with negative control group. Our findings indicated that polymer-based transfection may be a promising technique that allows the targeting of gene therapy for vein graft restenosis.
authors.affiliation: Department of Cardiothoracic Surgery, Second Affiliated Hospital, School of Medicine, Zhejiang University, Hangzhou 310009, China.
authors.forename: Xuebiao
authors.initials: X
authors.lastname: Li
authors.forename: Minjian
authors.initials: M
authors.lastname: Kong
authors.forename: Aiqiang
authors.initials: A
authors.lastname: Dong
authors.forename: Jianfang
authors.initials: J
authors.lastname: Qian
authors.forename: Daming
authors.initials: D
authors.lastname: Jiang
mesh_terms: D000818:Animals; D001011:Aorta / Q000166:cytology / Q000378:metabolism; D002478:Cells, Cultured; D020868:Gene Silencing; D015316:Genetic Therapy / Q000379:methods*; D006083:Graft Occlusion, Vascular / Q000517:prevention & control*; D007601:Jugular Veins / Q000637:transplantation; D009131:Muscle, Smooth, Vascular / Q000166:cytology / Q000378:metabolism; D010957:Plasmids; D011108:Polymers / Q000008:administration & dosage*; D034741:RNA, Small Interfering / Q000627:therapeutic use*; D011817:Rabbits; D017526:Receptor, IGF Type 1 / Q000096:biosynthesis / Q000235:genetics*; D014162:Transfection / Q000379:methods
chemical_list: D011108:Polymers; D034741:RNA, Small Interfering; D017526:Receptor, IGF Type 1
keywords: gene therapy; igf-1r; polymers; vein graft restenosis