重组腺相关病毒(rAAV)介导基因SMCKA3999对Duchenne肌营养不良病理和功能的影响

Recombinant adeno-associated virus vector carrying human minidystrophin gene SMCKA3999 effectively ameliorates dystrophic pathology and improves muscular function in mdx model mice

目的 研究重组腺相关病毒 (rAAV)载体介导的dystrophin小基因SMCKA3999治疗DMD模型鼠mdx ,从病理和功能观察rAAVSMCKA3999治疗对DMD模型小鼠mdx的疗效。方法 以dystrophin小基因SMCK A3999为目的基因 ,将SMCKA3999克隆至rAAV并包装成rAAVSMCKA3999,以 5× 10 10 病毒颗粒单点注射于DMD模型鼠mdx腓肠肌 ,基因治疗后 4个月及 7个月 ,采用免疫荧光、光镜组织病理、肌电图等方法 ,从形态和功能观察rAAVSMCKA3999治疗对DMD模型小鼠mdx的疗效。 结果 rAAVSMCKA3999使肌膜缺失的dys trophin恢复并稳定表达持续 7个月以上 ,肌肉组织病理改变好转 ,肌病肌电图改变明显改善 ,疗效持续 4个月以上。 结论 rAAVSMCKA3999能改善mdx小鼠骨骼肌的病理及功能 ,采用rAAV介导的dystrophin小基因SMC KA3999对Duchenne肌营养不良基因治疗是有希望的治疗方法。

Objective To study if recombinant adeno-associated virus vector(rAAV) mediated dystrophin minigene SMCKA3999 could effectively ameliorate dystrophic pathology and improve EMG in mdx model mice. Methods rAAV carrying SMCKA3999 was constructed and delivered into skeletal muscle of mdx mice by intramuscular injection. Immunofluorescent and HE stainng were adopted, and a needle was inserted into gastrocnemius muscle for recording EMG after 4 months of gene therapy. Results rAAVSMCK3999 resulted in efficient and stable restoring and expression of the missing dystrophin onto the plasma membrane. The expression persisted for 7 months. At the same time, rAAVSMCK3999 could improve pathologic changes and EMG of skeletal muscle in mdx mice. Conclusion The effectiveness of rAAVSMCKA3999 demonstrated in the study may offer a promising avenue of gene therapy for Duchenne muscular dystrophy.