Utilization of a novel Sendai virus vector in ex vivo gene therapy for hemophilia A
Utilization of a novel Sendai virus vector in ex vivo gene therapy for hemophilia A
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新型仙台病毒载体在甲型血友病离体基因治疗中的应用
DOI：
10.1007/s12185-020-03059-6
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发表时间：
2021
影响因子：
2.1
通讯作者：
Takada Hidetoshi
中科院分区：
文献类型：
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作者：
Yamaki Yuni;Fukushima Takashi;Yoshida Naomi;Nishimura Ken;Fukuda Aya;Hisatake Koji;Aso Masayuki;Sakasai Tomoki;Kijima-Tanaka Junko;Miwa Yoshihiro;Nakanishi Mahito;Sumazaki Ryo;Takada Hidetoshi
仙台病毒 (SeV) 载体被认为是基因转移的优质工具。在这里，我们使用近红外荧光蛋白（iRFP）介导的体内成像系统报告了新型、高性能、复制缺陷型和持久性仙台病毒（SeVdp）载体在培养细胞和小鼠中的转染功效。新型SeVdp载体建立了持续感染，并且插入基因的强表达在体外无限期持续。对皮下移植到 NOG、裸鼠和 ICR 小鼠中的 iRFP 表达细胞的分析表明，先天免疫参与了移植细胞的排除。我们还评估了这种新型 SeVdp 载体用于血友病 A 基因治疗的可行性。该系统能够插入全长 FVIII 基因，并且转导细胞将 FVIII 分泌到培养基中。腹膜内移植这些 FVIII 分泌细胞后，在小鼠血浆中检测到瞬时 FVIII 活性。逃避免疫的方法的进一步改进，例如免疫调节基因的同时表达，将使这种新型载体成为再生医学中非常有用的工具。
Sendai virus (SeV) vectors are being recognized as a superior tool for gene transfer. Here, we report the transfection efficacy of a novel, high-performance, replication-defective, and persistent Sendai virus (SeVdp) vector in cultured cells and in mice using a near-infrared fluorescent protein (iRFP)-mediated in vivo imaging system. The novel SeVdp vector established persistent infection, and strong expression of inserted genes was sustained indefinitely in vitro. Analysis of iRFP-expressing cells transplanted subcutaneously into NOG, nude, and ICR mice suggests that innate immunity was involved in the exclusion of the transplanted cells. We also evaluated the feasibility of this novel SeVdp vector for hemophilia A gene therapy. This system enabled insertion of full-length FVIII genes, and transduced cells secreted FVIII into the culture medium. Transient FVIII activity was detected in the plasma of mice after intraperitoneal transplantation of these FVIII-secreting cells. Further improvement in methods to evade immunity, such as simultaneous expression of immunomodulatory genes, would make this novel vector a very useful tool in regenerative medicine.