title: Synergistic effects of ultrasound-targeted microbubble destruction and TAT peptide on gene transfection: an experimental study in vitro and in vivo.
abstract: Cell-permeable peptides (CPPs) and ultrasound-targeted microbubble destruction (UTMD) have tremendous potential for gene delivery. However, their applications are limited due to nonspecificity of CPPs and low transfection efficiency of UTMD. Here, we developed a 'smart' gene delivery system by encapsulating TAT peptide (TATp) and hepatocyte growth factor (HGF) gene within lipid microbubbles, in which TATp was protected from being enzymatically cleaved and HGF gene was protected from degradation. This new strategy had synergistic effects of UTMD and TATp on gene transfection. We investigated the efficacy and safety of HGF gene transfection mediated by the combination of UTMD and TATp in vitro and in vivo. The results from MTT assay and flow cytometry analyses indicated that the combination of UTMD and TATp could enhance HGF gene expression in HUVECs without any significant side effect on cell viability. In rat myocardial infarction models, we demonstrated that the protein and mRNA expressions of HGF in myocardium caused by the combination of UTMD and TATp were the highest. Histopathological findings demonstrated that the combination of UTMD and TATp enhanced myocardial microvasculature and ameliorated myocardial fibrosis. In conclusion, the combination of UTMD and TATp might be a safe and efficient technique for gene delivery.
authors.affiliation: Department of Ultrasound, The Second Affiliated Hospital of Chongqing Medical University, Institute of Ultrasound Imaging of Chongqing Medical University, China.
authors.forename: Zhiyi
authors.initials: Z
authors.lastname: Zhou
authors.forename: Ping
authors.initials: P
authors.lastname: Zhang
authors.forename: Jianli
authors.initials: J
authors.lastname: Ren
authors.forename: Haitao
authors.initials: H
authors.lastname: Ran
authors.forename: Yuanyi
authors.initials: Y
authors.lastname: Zheng
authors.forename: Pan
authors.initials: P
authors.lastname: Li
authors.forename: Qunxia
authors.initials: Q
authors.lastname: Zhang
authors.forename: Maohui
authors.initials: M
authors.lastname: Zhang
authors.forename: Zhigang
authors.initials: Z
authors.lastname: Wang
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D003094:Collagen / Q000378:metabolism; D004247:DNA; D049452:Green Fluorescent Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D017228:Hepatocyte Growth Factor / Q000737:chemistry / Q000235:genetics; D061307:Human Umbilical Vein Endothelial Cells / Q000378:metabolism; D006801:Humans; D008297:Male; D045423:Microbubbles; D009203:Myocardial Infarction / Q000378:metabolism / Q000628:therapy*; D009206:Myocardium / Q000378:metabolism; D010455:Peptides / Q000008:administration & dosage* / Q000737:chemistry; D010957:Plasmids; D051381:Rats; D017207:Rats, Sprague-Dawley; D013010:Sonication; D014162:Transfection / Q000379:methods*; D054322:tat Gene Products, Human Immunodeficiency Virus / Q000737:chemistry*
chemical_list: D018952:Antigens, CD34; D010455:Peptides; C485184:enhanced green fluorescent protein; D054322:tat Gene Products, Human Immunodeficiency Virus; D049452:Green Fluorescent Proteins; D017228:Hepatocyte Growth Factor; D003094:Collagen; D004247:DNA
keywords: Cell-permeable peptide; Gene transfection; Myocardial infarction; Ultrasound-targeted microbubble destruction