Adeno-associated viral vector mediated and cardiac-specific delivery of CD151 gene in ischemic rat hearts

Abstract

Our previous studies demonstrated that CD151 gene promoted neovascularization in ischemic heart model. To improve the delivery efficacy and target specificity of CD151 gene to ischemic heart, we generated an adeno-associated virus (AAV) vector in which CD151 expression was controlled by the myosin light chain (MLC-2v) promoter to achieve the cardiac-specific expression of CD151 gene in ischemic myocardium and to limit unwanted CD151 expression in extracardiac organs. The function of this vector was examined in rat ischemic myocardium model. The protein expression of CD151 in the ischemic myocardium areas, liver and kidney was confirmed by using Western blot, while the microvessels within ischemic myocardium areas were detected by using immunohistochemistry. The results showed that MLC-2v significantly enhanced the expression of CD151 in ischemic myocardium, but attenuated its expression in other organs. The forced CD151 expression could increase the number of microvessels in the ischemic myocardium. This study demonstrates the AAV-mediated and MLC-2v regulated CD151 gene is highly expressed in the ischemic myocardium and cardiac-specific delivery that is more efficiently targets CD151 to the ischemia myocardium after myocardial infarction.

Keywords

cardiac ischemia / CD151 / angiogenesis / gene therapy / MLC-2v

References

[1] LanR.F., LiuZ.X., LiuX.C., et al.. CD151 promotes neovascularization and improves blood perfusion in a rat hind-limb ischemia model. J Endovasc Ther, 2005, 12(4): 469-478

[2] ZhengZ., LiuZ.. CD151 gene delivery activates PI3K/Akt pathway and promotes neovascularization after myocardial infarction in rats. Mol Med, 2006, 12(9–10): 214-220

[3] ZuoH., LiuZ., LiuX., et al.. CD151 gene delivery after myocardial infarction promotes functional neovascularization and activates FAK signaling. Mol Med, 2009, 15(9–10): 307-315

[4] ZuoH.J., LiuZ.X., LiuX.C., et al.. Assessment of myocardial blood perfusion improved by CD151 in a pig myocardial infarction model. Acta Pharmacol Sin, 2009, 30(1): 70-77

[5] LeeR.J., SpringerM.L., Blanco-BoseW.E., et al.. VEGF gene delivery to myocardium: deleterious effects of unregulated expression. Circulation, 2000, 102(8): 898-901

[6] ShenF., FanY., SuH., et al.. Adeno-associated viral vector-mediated hypoxia-regulated VEGF gene transfer promotes angiogenesis following focal cerebral ischemia in mice. Gene Ther, 2008, 15(1): 30-39

[7] PhillipsM.I., TangY., Schmidt-OttK., et al.. Vigilant vector: heart-specific promoter in an adeno-associated virus vector for cardioprotection. Hypertension, 2002, 39(2): 651-655

[8] LanR., LiuZ., SongY., et al.. Effects of rAAV-CD151 and rAAV-antiCD151 on the migration of human tongue squamous carcinoma cell line Tca8113. J Huazhong Univ Sci Technol [Med Sci], 2004, 24(6): 556-559

[9] XiaJ.L., DaiC., MichalopoulosG.K., et al.. Hepatocyte growth factor attenuates liver fibrosis induced by bile duct ligation. Am J Pathol, 2006, 168(5): 1500-1512

[10] DengX., SzaboS., KhomenkoT., et al.. Gene therapy with adenoviral plasmids or naked DNA of vascular endothelial growth factor and platelet-derived growth factor accelerates healing of duodenal ulcer in rats. J Pharmacol Exp Ther, 2004, 311(3): 982-988

[11] SimonsM., AnnexB.H., LahamR.J., et al.. Pharmacological treatment of coronary artery disease with recombinant fibroblast growth factor-2: double-blind, randomized, controlled clinical trial. Circulation, 2002, 105(7): 788-793

[12] LahamR.J., ChronosN.A., PikeM., et al.. Intracoronary basic fibroblast growth factor (FGF-2) in patients with severe ischemic heart disease: results of a phase I open-label dose escalation study. J Am Coll Cardiol, 2000, 36(7): 2132-2139

[13] SuH., LuR., KanY.W.. Adeno-associated viral vector-mediated vascular endothelial growth factor gene transfer induces neovascular formation in ischemic heart. Proc Natl Acad Sci U S A, 2000, 97(25): 13801-13 806

[14] SungH.K., KimY.W., ChoiS.J., et al.. COMP-angiopoietin-1 enhances skeletal muscle blood flow and insulin sensitivity in mice. Am J Physiol Endocrinol Metab, 2009, 297(2): E402-409

[15] HasegawaH., UtsunomiyaY., KishimotoK., et al.. SFA-1, a novel cellular gene induced by human T-cell leukemia virus type 1, is a member of the transmembrane 4 superfamily. J Virol, 1996, 70(5): 3258-3263

[16] FitterS., TetazT.J., BerndtM.C., et al.. Molecular cloning of cDNA encoding a novel platelet-endothelial cell tetra-span antigen, PETA-3. Blood, 1995, 86(4): 1348-1355

[17] SincockP.M., FitterS., PartonR.G., et al.. PETA-3/CD151, a member of the transmembrane 4 superfamily, is localized to the plasma membrane and endocytic system of endothelial cells, associates with multiple integrins and modulates cell function. J Cell Sci, 1999, 112(Pt6): 833-844

[18] YangJ., LiuZ., ShenX., et al.. Expression of CD151 in human atherosclerotic artery and its implication. J Huazhong Univ Sci Technol [Med Sci], 2005, 25(6): 629-631

[19] LiuL., HeB., LiuW.M., et al.. Tetraspanin CD151 promotes cell migration by regulating integrin trafficking. J Biol Chem, 2007, 282(43): 31 631-31 642

[20] ZhangX.A., KazarovA.R., YangX., et al.. Function of the tetraspanin CD151-alpha6beta1 integrin complex during cellular morphogenesis. Mol Biol Cell, 2002, 13(1): 1-11

[21] TakedaY., KazarovA.R., ButterfieldC.E., et al.. Deletion of tetraspanin Cd151 results in decreased pathologic angiogenesis in vivo and in vitro. Blood, 2007, 109(4): 1524-1532

[22] ZhengZ.Z., LiuZ.X.. Activation of the phosphatidy linositol 3-kinase/protein kinase Akt pathway mediates CD151-induced endothelial cell proliferation and cell migration. Int J Biochem Cell Biol, 2007, 39(2): 340-348

[23] ShenF., SuH., FanY., et al.. Adeno-associated viral-vector-mediated hypoxia-inducible vascular endothelial growth factor gene expression attenuates ischemic brain injury after focal cerebral ischemia in mice. Stroke, 2006, 37(10): 2601-2606

[24] SchwarzE.R., SpeakmanM.T., PattersonM., et al.. Evaluation of the effects of intramyocardial injection of DNA expressing vascular endothelial growth factor (VEGF) in a myocardial infarction model in the rat-angiogenesis and angioma formation. J Am Coll Cardiol, 2000, 35(5): 1323-1330

[25] SuH., JohoS., HuangY., et al.. Adeno-associated viral vector delivers cardiac-specific and hypoxia-inducible VEGF expression in ischemic mouse hearts. Proc Natl Acad Sci U S A, 2004, 101(46): 16 280-16 285

[26] SmallE.M., KriegP.A.. Molecular regulation of cardiac chamber-specific gene expression. Trends Cardiovasc Med, 2004, 14(1): 13-18