Gene therapy for hemophilia B mice with scAAV8-LP1-hFIX

Abstract

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-associated 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 liver-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-LP1-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 5×10¹¹ and 1×10¹¹ 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.

Keywords

hemophilia B / AAV8 / hFIX / gene therapy

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Acknowledgements

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).

Compliance with ethics guidelines

Wei Lu, Qingzhang Zhou, Hao Yang, Hao Wang, Yexing Gu, Qi Shen, Jinglun Xue, Xiaoyan Dong, and Jinzhong Chen declare no conflict of interest. All animal procedures were performed in accordance with institutional and national guidelines for the care and use of laboratory animals.

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2016 Higher Education Press and Springer-Verlag Berlin Heidelberg