Targeted introduction of tissue plasminogen activator (TPA) at the AAVS1 locus in mesenchymal stem cells (MSCs) and its stable and effective expression.
Targeted introduction of tissue plasminogen activator (TPA) at the AAVS1 locus in mesenchymal stem cells (MSCs) and its stable and effective expression.
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DOI：
10.1016/j.bbrc.2013.06.037
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发表时间：
2013-07
影响因子：
3.1
通讯作者：
Shu-Jun Li;Rui-zheng Shi;Yongping Bai;Dan Hong;Wei Yang;Xiang Wang;L. Mo;Guogang Zhang
中科院分区：
文献类型：
--
作者：
Shu-Jun Li;Rui-zheng Shi;Yongping Bai;Dan Hong;Wei Yang;Xiang Wang;L. Mo;Guogang Zhang
组织型纤溶酶原激活剂（TPA）是治疗急性心肌梗死的有效方法。然而，TPA的全身给药与出血风险相关。骨髓间充质干细胞（Mesenchymal stem cells，MSCs）具有免疫原性低、可向损伤组织归巢等特点，是实现靶向治疗的理想细胞载体。本文将TPA基因整合到骨髓间充质干细胞的AAVS 1中，证实其是一个安全的染色体位点。靶向效率为83%。具有位点特异性整合的克隆保留了MSC的干细胞特性，显示正常的核型，并且可以持续有效地表达TPA，如通过1.5单位/mL的平均表达活性（是对照组的3.4倍）所证明的。在继代培养和随后的生长两周后，克隆显示出1.43单位/mL的平均TPA活性，并且在各个克隆之间没有表现出显著差异。总之，外源TPA基因可以被特异性地引入AAVS 1基因座，从而它可以稳定和有效地表达。MSC可以用作使用TPA靶向治疗血栓的细胞载体。
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.