Lentivirus-mediated platelet-derived factor VIII gene therapy in murine haemophilia A
Lentivirus-mediated platelet-derived factor VIII gene therapy in murine haemophilia A
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DOI：
10.1111/j.1538-7836.2007.02346.x
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发表时间：
2007-02-01
影响因子：
10.4
通讯作者：
Montgomery, R. R.
中科院分区：
文献类型：
--
作者：
Shi, Q.;Wilcox, D. A.;Montgomery, R. R.
背景资料：我们实验室以前的研究已经证明，在血小板特异性整合素α IIb基因启动子（2bF 8）的指导下，因子VIII（FVIII）的谱系靶向合成可以纠正小鼠血友病A表型，即使在转基因小鼠模型中存在高滴度抑制性抗体。目的：在这项研究中，我们评估了使用基因治疗方法，以纠正血友病A的FVIII缺陷（FVIIInull）小鼠的骨髓（BM）移植与慢病毒（LV）为基础的基因转移盒编码2bF 8的疗效。结果如下：在处理小鼠的血小板裂解物中检测到功能性FVIII活性（FVIII：C），其水平与2bF 8杂合转基因小鼠相似。移植有2bF 8 LV转导的BM的小鼠在剪尾后存活，并且在本研究期间（11个月），我们没有检测到抑制性或非抑制性FVIII抗体。此外，从初次移植受者转移到FVIIInull二次移植受者的BM显示持续的血小板-FVIII表达，导致血友病A表型的纠正，表明基因转移发生在长期重建的造血干细胞内。结论：这些结果表明，通过慢病毒介导的骨髓转导/移植在血小板中异位表达FVIII可能是人类血友病A基因治疗的一种有前途的策略。
Background: Previous studies from our laboratory have demonstrated that lineage-targeted synthesis of factor VIII (FVIII) under the direction of the platelet-specific integrin alpha IIb gene promoter (2bF8) can correct the murine haemophilia A phenotype even in the presence of high titer inhibitory antibodies in a transgenic mouse model. Objective: In this study, we assessed the efficacy of using a genetic therapy approach to correct haemophilia A in FVIII-deficient (FVIIInull) mice by transplantation of bone marrow (BM) transduced with a lentivirus (LV)-based gene transfer cassette encoding 2bF8. Results: Functional FVIII activity (FVIII:C) was detected in platelet lysates from treated mice and the levels were similar to 2bF8 heterozygous transgenic mice. Mice transplanted with 2bF8 LV-transduced BM survived tail clipping and we did not detected inhibitory or non-inhibitory FVIII antibodies over the period of this study (11 months). Furthermore, BM transferred from the primary transplant recipients into FVIIInull secondary recipients demonstrated sustained platelet-FVIII expression leading to correction of the haemophilia A phenotype showing that gene transfer occurred within long-term repopulating haematopoietic stem cells. Conclusions: These results demonstrate that ectopic expression of FVIII in platelets by lentivirus-mediated bone marrow transduction/transplantation may be a promising strategy for gene therapy of haemophilia A in humans.