An induced pluripotent stem cell-mediated and integration-free factor VIII expression system.
An induced pluripotent stem cell-mediated and integration-free factor VIII expression system.
复制标题
DOI：
10.1016/j.bbrc.2012.12.096
复制
发表时间：
2013-02
影响因子：
3.1
通讯作者：
Y. Yakura;Chie Ishihara;Hajime Kurosaki;Y. Kazuki;N. Komatsu;Yoshiaki Okada;T. Doi;Hiroyuki Takeya;M. Oshimura
中科院分区：
文献类型：
--
作者：
Y. Yakura;Chie Ishihara;Hajime Kurosaki;Y. Kazuki;N. Komatsu;Yoshiaki Okada;T. Doi;Hiroyuki Takeya;M. Oshimura
人类人工染色体(HAC)作为基因治疗载体具有许多优点，包括稳定的上体维持和携带大基因插入的能力。诱导多能干细胞(IPS)在基因治疗方面也有很大的潜力，它可以从个人自身的组织中产生，并在重新引入时对任何组织都有贡献。带有重编程因子的仙台病毒(Sendai Virus，SeV)载体是产生iPS细胞的有力工具，因为它具有高感染效率而不存在整合到宿主染色体的风险。在本研究中，我们利用非整合的SeV和HAC载体，建立了一种iPS细胞介导的无整合凝血因子VIII(FVIII)的表达系统。多个人FVIII基因被插入到HAC载体(PF4-FVIII-HAc)中，这些基因由巨核细胞特异性血小板因子-4(PF4)启动子控制，用于血友病A的治疗。将pF4-FVIII-HAC导入SeV载体介导的来源于血友病A小鼠模型的iPS细胞，体外诱导iPS细胞分化为巨核细胞/血小板后，pF4-FVIII-HAC诱导表达FVIII。本研究利用两个非整合载体构建了iPS细胞介导的PF4驱动的FVIII表达系统；因此，该系统有望成为血友病A的更安全的基因和细胞治疗工具。
Human artificial chromosome (HAC) has several advantages as a gene therapy vector, including stable episomal maintenance and the ability to carry large gene inserts. Induced pluripotent stem (iPS) cells also have a great potential for gene therapy, which can be generated from an individual’s own tissues and contribute to any tissues when reintroduced. A Sendai virus (SeV) vector with reprogramming factors is a powerful tool for generating iPS cells because of the high infection efficiency without the risk of integration into host chromosomes. In this study, we developed an iPS cell-mediated and integration-free coagulation factor VIII (FVIII) expression system using non-integrating SeV- and HAC-vectors. Multiple human FVIII genes, which were under the control of the megakaryocyte-specific platelet factor-4 (PF4) promoter for development of a treatment for hemophilia A, were inserted into a HAC vector (PF4-FVIII-HAC). The PF4-FVIII-HAC was introduced into SeV vector-mediated iPS cells derived from a mouse model of hemophilia A. After in vitro differentiation of iPS cells with the PF4-FVIII-HAC into megakaryocytes/platelets, the PF4-FVIII-HAC resulted in expression of FVIII. This study has developed the iPS cell-mediated PF4-driven FVIII expression system using two non-integrating vectors; therefore, this system may be a promising tool for safer gene- and cell-therapy of hemophilia A.