Long-term correction of hemorrhagic diathesis in hemophilia A mice by an AAV-delivered hybrid FVIII composed of the human heavy chain and the rat light chain
Long-term correction of hemorrhagic diathesis in hemophilia A mice by an AAV-delivered hybrid FVIII composed of the human heavy chain and the rat light chain
复制标题
通过 AAV 递送的由人重链和大鼠轻链组成的杂交 FVIII 对血友病 A 小鼠的出血素质进行长期纠正
作者：
J. Mao;Yun Wang;Wei Zhang;Yan Shen;Guowei Zhang;Wenda Xi;Qiang Wang;Z. Ruan;Jin Wang;X. Xi
血友病A （HA）的常规治疗是预防性或按需静脉注射FVIII。然而，它们既昂贵又不方便。因此，必须制定更好的HA治疗策略。在这项研究中，开发了一种重组FVIII cDNA，编码具有增强促凝潜能的人/大鼠杂交FVIII，用于腺相关病毒（AAV）传递的基因治疗。将含有人FVIII重链（hHC）、人FVIII轻链（hLC）和大鼠FVIII轻链（rLC）的质粒转染到细胞中，并以流体动力学方式注射到HA小鼠体内。纯化的AAV病毒以两种剂量静脉注射到HA小鼠体内。结果表明，在相同表达水平下，hHC + rLC蛋白比hHC + hLC蛋白具有更高的活性。hHC + rLC的比活性比其对应体高4 ~ 8倍。流体动力注射实验得到了一致的结果。值得注意的是，接受aav递送的hHC + rLC治疗的HA小鼠的活性明显高于hHC + hLC治疗的小鼠，并且治疗效果持续长达40周。综上所述，通过aav传递基因治疗，应用hHC + rLC杂交FVIII，可显著改善HA小鼠的出血素质。这些数据可能有助于开发用于HA基因治疗的FVIII表达盒。
Conventional therapies for hemophilia A (HA) are prophylactic or on-demand intravenous FVIII infusions. However, they are expensive and inconvenient to perform. Thus, better strategies for HA treatment must be developed. In this study, a recombinant FVIII cDNA encoding a human/rat hybrid FVIII with an enhanced procoagulant potential for adeno-associated virus (AAV)-delivered gene therapy was developed. Plasmids containing human FVIII heavy chain (hHC), human light chain (hLC), and rat light chain (rLC) were transfected into cells and hydrodynamically injected into HA mice. Purified AAV viruses were intravenously injected into HA mice at two doses. Results showed that the hHC + rLC protein had a higher activity than the hHC + hLC protein at comparable expression levels. The specific activity of hHC + rLC was about 4- to 8-fold higher than that of their counterparts. Hydrodynamic injection experiments obtained consistent results. Notably, the HA mice undergoing the AAV-delivered hHC + rLC treatment exhibited a visibly higher activity than those treated with hHC + hLC, and the therapeutic effects lasted for up to 40 weeks. In conclusion, the application of the hybrid FVIII (hHC + rLC) via an AAV-delivered gene therapy substantially improved the hemorrhagic diathesis of the HA mice. These data might be of help to the development of optimized FVIII expression cassette for HA gene therapy.
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影响因子：
20.3
作者：
McIntosh, Jenny;Lenting, Peter J.;Nathwani, Amit C.
通讯作者：
Nathwani, Amit C.
人/猪混合因子 VIII 分子的凝血特性。
DOI：
--
发表时间：
1992
期刊：
The Journal of biological chemistry
影响因子：
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作者：
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通讯作者：
Fay,PJ
影响因子：
20.3
作者：
O'Brien, LM;Mastri, M;Fay, PJ
通讯作者：
Fay, PJ
影响因子：
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作者：
Wang, Qizhao;Dong, Biao;Xiao, Weidong
通讯作者：
Xiao, Weidong
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DOI：
10.1016/j.bcmd.2018.09.004
发表时间：
2018
期刊：
Blood cells, molecules & diseases
影响因子：
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作者：
Zhang,Wei;Mao,Jianhua;Shen,Yan;Zhang,Guowei;Shao,Yanyan;Ruan,Zheng;Wang,Yun;Wu,Wenman;Wang,Xuefeng;Zhu,Jiang;Chen,Saijuan;Xiao,Weidong;Xi,Xiaodong
通讯作者：
Xi,Xiaodong