Ectopic Expression of FVIII in HPCs and MSCs Derived from hiPSCs with Site-Specific Integration of ITGA2B Promoter-Driven BDDF8 Gene in Hemophilia A.
Ectopic Expression of FVIII in HPCs and MSCs Derived from hiPSCs with Site-Specific Integration of ITGA2B Promoter-Driven BDDF8 Gene in Hemophilia A.
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A 型血友病中 ITGA2B 启动子驱动的 BDDF8 基因位点特异性整合的 hiPSC 衍生的 HPC 和 MSC 中 FVIII 的异位表达
DOI：
10.3390/ijms23020623
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发表时间：
2022-01-06
影响因子：
5.6
通讯作者：
Liang D
中科院分区：
文献类型：
--
作者：
Zhao J;Zhou M;Wang Z;Wu L;Hu Z;Liang D
血友病A （HA）是由凝血因子VIII （FVIII）基因（F8）突变引起的。基因疗法有望治愈血凝素；然而，FVIII抑制剂的形成阻碍了其临床应用。鉴于血小板通过局部释放α-颗粒促进凝血，FVIII在血小板中异位表达已被尝试，并在HA治疗中取得了良好的结果。在截断的ITGA2B启动子驱动下，b结构域缺失的F8 （BDDF8）靶向HA患者特异性诱导多能干细胞（HA- ipscs）的核糖体DNA （rDNA）位点。将FVIII修饰的人诱导多能干细胞（2bF8-iPSCs）分化为诱导造血祖细胞（iHPCs）、诱导巨核细胞（iMKs）和间充质干细胞（iMSCs），并检测FVIII的表达。ITGA2B启动子驱动的BDDF8位点特异性整合到HA-iPSCs的rDNA位点。2bF8-iPSCs被高效分化为2bF8-iHPCs、2bF8-iMKs和2bF8-iMSCs。在2bF8-iHPCs的裂解物中，FVIII为10.31 ng/106个细胞，而在HA-iHPCs中为1.56 ng/106个细胞，在2bF8-iMSCs裂解物中，FVIII为3.64 ng/106个细胞，而在cmv驱动的BDDF8的iMSCs中，FVIII为1.31 ng/106个细胞。我们的研究结果表明，在ITGA2B启动子驱动的BDDF8位点特异性整合的hipsc衍生的iHPCs和iMSCs中，FVIII高表达，表明这种靶向血小板的HA基因治疗策略具有潜在的临床前景。
Hemophilia A (HA) is caused by mutations in the coagulation factor VIII (FVIII) gene (F8). Gene therapy is a hopeful cure for HA; however, FVIII inhibitors formation hinders its clinical application. Given that platelets promote coagulation via locally releasing α-granule, FVIII ectopically expressed in platelets has been attempted, with promising results for HA treatment. The B-domain-deleted F8 (BDDF8), driven by a truncated ITGA2B promoter, was targeted at the ribosomal DNA (rDNA) locus of HA patient-specific induced pluripotent stem cells (HA-iPSCs). The F8-modified, human induced pluripotent stem cells (2bF8-iPSCs) were differentiated into induced hematopoietic progenitor cells (iHPCs), induced megakaryocytes (iMKs), and mesenchymal stem cells (iMSCs), and the FVIII expression was detected. The ITGA2B promoter-driven BDDF8 was site-specifically integrated into the rDNA locus of HA-iPSCs. The 2bF8-iPSCs were efficiently differentiated into 2bF8-iHPCs, 2bF8-iMKs, and 2bF8-iMSCs. FVIII was 10.31 ng/106 cells in lysates of 2bF8-iHPCs, compared to 1.56 ng/106 cells in HA-iHPCs, and FVIII was 3.64 ng/106 cells in 2bF8-iMSCs lysates, while 1.31 ng/106 cells in iMSCs with CMV-driven BDDF8. Our results demonstrated a high expression of FVIII in iHPCs and iMSCs derived from hiPSCs with site-specific integration of ITGA2B promoter-driven BDDF8, indicating potential clinical prospects of this platelet-targeted strategy for HA gene therapy.
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一项对患有 A 型血友病的狗进行 AAV 基因治疗的长期研究确定了转导肝细胞的克隆扩增。
影响因子：
46.9
作者：
Nguyen GN;Everett JK;Kafle S;Roche AM;Raymond HE;Leiby J;Wood C;Assenmacher CA;Merricks EP;Long CT;Kazazian HH;Nichols TC;Bushman FD;Sabatino DE
通讯作者：
Sabatino DE
DOI：
10.1111/jth.12750
发表时间：
2014-12
期刊：
Journal of thrombosis and haemostasis : JTH
影响因子：
--
作者：
Fomin ME;Togarrati PP;Muench MO
通讯作者：
Muench MO
影响因子：
10.4
作者：
Liu, X.;Liu, M.;Xia, J.
通讯作者：
Xia, J.
影响因子：
10.4
作者：
Shi, Q.;Wilcox, D. A.;Montgomery, R. R.
通讯作者：
Montgomery, R. R.
影响因子：
20.3
作者：
Takayama, Naoya;Nishikii, Hidekazu;Nakauchi, Hirornitsu
通讯作者：
Nakauchi, Hirornitsu