CRISPR /Cas9‐mediated somatic correction of a novel coagulator factor IX gene mutation ameliorates hemophilia in mouse

• DOI: 10.15252/emmm.201506039
• Publication Date: 2016-03-11T02:09:51Z
• Authors: Yuting Guan; Yanlin Ma; Qi Li; Zhenliang Sun; Lie Ma; Lijuan Wu; Liren Wang; Li Zeng; Yanjiao Shao; Yuting Chen; Ning Ma; Wenqing Lu; Kewen Hu; Honghui Han; Yanhong Yu; Yuanhua Huang; Mingyao Liu; Dali Li
• Keywords: Male; 0301 basic medicine; Medicine (General); Genetic Vectors; Mutation, Missense; QH426-470; Hemophilia B; Adenoviridae; Factor IX; Mice; 03 medical and health sciences; R5-920; Genetics; genome editing; monogenetic disease; Animals; Humans; Child; Research Articles; Gene Editing; Recombination, Genetic; Genetic Therapy; gene therapy; 3. Good health; Disease Models, Animal; Treatment Outcome; hemostasis; hemophilia B; CRISPR-Cas Systems

The X-linked genetic bleeding disorder caused by deficiency of coagulator factor IX, hemophilia B, is a disease ideally suited for gene therapy with genome editing technology. Here, we identify a family with hemophilia B carrying a novel mutation, Y371D, in the human F9 gene. The CRISPR/Cas9 system was used to generate distinct genetically modified mouse models and confirmed that the novel Y371D mutation resulted in a more severe hemophilia B phenotype than the previously identified Y371S mutation. To develop therapeutic strategies targeting this mutation, we subsequently compared naked DNA constructs versus adenoviral vectors to deliver Cas9 components targeting the F9 Y371D mutation in adult mice. After treatment, hemophilia B mice receiving naked DNA constructs exhibited correction of over 0.56% of F9 alleles in hepatocytes, which was sufficient to restore hemostasis. In contrast, the adenoviral delivery system resulted in a higher corrective efficiency but no therapeutic effects due to severe hepatic toxicity. Our studies suggest that CRISPR/Cas-mediated in situ genome editing could be a feasible therapeutic strategy for human hereditary diseases, although an efficient and clinically relevant delivery system is required for further clinical studies.