Gene therapy for hemophilia B mice with scAAV8-LPI-hFIX  

作　　者：Wei Lu Qingzhang Zhou Hao Yang Hao Wang Yexing Gu Qi Shen Jinglun Xue Xiaoyan Dong Jinzhong Chen 

机构地区：[1]State Key Laboratory of Genetic Engineering, Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China [2]Beijing Five Plus Molecular Medicine Institute, Beijing 100176, China [3]College of Life Sciences, Jilin University, Changchun 130012, China

出　　处：《Frontiers of Medicine》2016年第2期212-218,共7页医学前沿（英文版）

基　　金：This work was supported by the National Natural Science Foundation of China （Nos. 31071171 and 81170532） and the National High Technology Research and Development Program of China （863 Program, No. 2012AA020810）.

摘　　要：Hemophilia B is a hemorrhagic disease caused by the deficiency of clotting factor IX （FIX）. Gene therapy might be the ultimate strategy for the disease. However, two main problems that should be solved in gene therapy for hemophilia B are immunity and safety. Self-complementary adeno-assoeiated virus serotype 8 （scAAV8）, a non-human primate AAV featuring low immunogenicity and high transfection efficiency in liver cells, might be a potential vector for hemophilia B gene therapy. A strong fiver-specific promoter-1 （LP1） was inserted and mutant human FIX Arg338Ala was introduced into plasmid scAAV8-LP1 to develop an optimized AAV8 vector that expresses human clotting factor FIX （hFIX）. The efficiency of scAAV8-LPI-hFIX administered through normal systemic injection or hydrodynamic injection was compared. A high expression was achieved using hydrodynamic injection, and the peak hFIX expression levels in the 5 × 10^11 and 1 × 10^11 virus genome （vg） cohorts were 31.94% and 25.02% of normal level, respectively, at 60 days post-injection. From the perspective of long-term （200 days） expression, both injection methods presented promising results with the concentration value maintained above 4% of normal plasma. The results were further verified by enzyme-linked immunosorbent assay and activated partial thromboplastin time. Our study provides a potential gene therapy method for hemophilia B.Hemophilia B is a hemorrhagic disease caused by the deficiency of clotting factor IX （FIX）. Gene therapy might be the ultimate strategy for the disease. However, two main problems that should be solved in gene therapy for hemophilia B are immunity and safety. Self-complementary adeno-assoeiated virus serotype 8 （scAAV8）, a non-human primate AAV featuring low immunogenicity and high transfection efficiency in liver cells, might be a potential vector for hemophilia B gene therapy. A strong fiver-specific promoter-1 （LP1） was inserted and mutant human FIX Arg338Ala was introduced into plasmid scAAV8-LP1 to develop an optimized AAV8 vector that expresses human clotting factor FIX （hFIX）. The efficiency of scAAV8-LPI-hFIX administered through normal systemic injection or hydrodynamic injection was compared. A high expression was achieved using hydrodynamic injection, and the peak hFIX expression levels in the 5 × 10^11 and 1 × 10^11 virus genome （vg） cohorts were 31.94% and 25.02% of normal level, respectively, at 60 days post-injection. From the perspective of long-term （200 days） expression, both injection methods presented promising results with the concentration value maintained above 4% of normal plasma. The results were further verified by enzyme-linked immunosorbent assay and activated partial thromboplastin time. Our study provides a potential gene therapy method for hemophilia B.

关 键 词：hemophilia B AAV8 hFIX gene therapy 

分 类 号：Q785[生物学—分子生物学] Q78