title: Targeted introduction of tissue plasminogen activator (TPA) at the AAVS1 locus in mesenchymal stem cells (MSCs) and its stable and effective expression.
abstract: Thrombolytic therapy using tissue plasminogen activator (TPA) is an effective method for treating acute myocardial infarction. However, the systemic administration of TPA is associated with the risk of hemorrhage. Mesenchymal stem cells (MSCs) from bone marrow are characterized by low immunogenicity and homing toward damaged tissues and are therefore ideal cell carriers to achieve lesion-targeting medication. In this article, TPA gene was integrated into the AAVS1 of mesenchymal stem cells, which has been confirmed to be a safe chromosomal locus. The targeting efficiency was 83%. The clones with the site-specific integration retained the stem cell traits of MSCs, displayed a normal karyotype and could persistently and effectively express TPA, as demonstrated by an average expression activity of 1.5 units/mL (3.4-fold that of the control group). After subculture and subsequent growth for two weeks, the clones showed an average TPA activity of 1.43 units/mL and exhibited no significant differences among the individual clones. In summary, the foreign TPA gene can be specifically introduced to the AAVS1 locus, whereby it can be stably and effectively expressed. MSCs can serve as cell carriers for the targeted treatment of a thrombus using TPA.
authors.affiliation: Department of Cardiovascular Medicine, Xiangya Hospital, Central South University, Changsha 410078, China.
authors.forename: Shu-Jun
authors.initials: SJ
authors.lastname: Li
authors.forename: Rui-Zheng
authors.initials: RZ
authors.lastname: Shi
authors.forename: Yong-Ping
authors.initials: YP
authors.lastname: Bai
authors.forename: Dan
authors.initials: D
authors.lastname: Hong
authors.forename: Wei
authors.initials: W
authors.lastname: Yang
authors.forename: Xiang
authors.initials: X
authors.lastname: Wang
authors.forename: Long
authors.initials: L
authors.lastname: Mo
authors.forename: Guo-Gang
authors.initials: GG
authors.lastname: Zhang
mesh_terms: D002999:Clone Cells; D015870:Gene Expression; D018390:Gene Targeting; D056426:Genetic Loci / Q000235:genetics*; D005822:Genetic Vectors / Q000235:genetics; D006801:Humans; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D010959:Tissue Plasminogen Activator / Q000235:genetics*
chemical_list: D010959:Tissue Plasminogen Activator
keywords: AAVS1; Gene targeting; Mesenchymal stem cells (MSCs); TPA; Thrombus; Zinc-finger nuclease (ZFN)