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

doi:10.1007/s11684-021-0844-7

Frontiers of Medicine

2022, Vol. 16

Issue (4): 584-595 https://doi.org/10.1007/s11684-021-0844-7

本期目录

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

Jianhua Mao¹(

), Yun Wang^1,², Wei Zhang¹, Yan Shen³, Guowei Zhang⁴, Wenda Xi⁵, Qiang Wang¹, Zheng Ruan¹, Jin Wang², Xiaodong Xi¹(

)

¹. Shanghai Institute of Hematology, State Key Laboratory of Medical Genomics, Collaborative Innovation Center of Hematology, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
². Shanghai Institute of Hematology, State Key Laboratory for Medical Genomics and Department of Hematology, Collaborative Innovation Center of Systems Biomedicine, Pôle Sino-Français des Sciences du Vivant et Genomique, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
³. Research Center for Experimental Medicine, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China
⁴. The School of Medicine, Hangzhou Normal University, Hangzhou 310036, China
⁵. Shanghai Institute of Hypertension, Ruijin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine, Shanghai 200025, China

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Abstract：

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.

Key words： hemophilia A adeno-associated virus (AAV) human/rat hybrid factor VIII gene therapy dual chain strategy

收稿日期: 2020-07-20 出版日期: 2022-09-02

Corresponding Author(s): Jianhua Mao,Xiaodong Xi

引用本文:

. [J]. Frontiers of Medicine, 2022, 16(4): 584-595.
Jianhua Mao, Yun Wang, Wei Zhang, Yan Shen, Guowei Zhang, Wenda Xi, Qiang Wang, Zheng Ruan, Jin Wang, Xiaodong Xi. 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. Front. Med., 2022, 16(4): 584-595.

链接本文:

https://academic.hep.com.cn/fmd/CN/10.1007/s11684-021-0844-7
https://academic.hep.com.cn/fmd/CN/Y2022/V16/I4/584

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