不同注射条件下AAV9载体对小鼠肌肉的短期转导效果  

Short-term transduction effect of AAV9 vector on mouse muscle under different injection conditions

作　　者：化春晓 薛淑文 蔡奥捷 李倩倩[1] 杨宇霞[1] 孔祥东[1] HUA Chunxiao;XUE Shuwen;CAI Aojie;LI Qianqian;YANG Yuxia;KONG Xiangdong(Genetics and Prenatal Diagnosis Center,The First Affiliated Hospital of Zhengzhou University,Zhengzhou 450052,China)

机构地区：[1]郑州大学第一附属医院遗传与产前诊断中心,河南郑州450052

出　　处：《空军军医大学学报》2024年第6期634-638,共5页Journal of Air Force Medical University

基　　金：国家自然科学基金(82000321);河南省医学科技攻关计划项目(LHGJ20190113)。

摘　　要：目的观察不同注射方式和注射剂量下腺相关病毒9型(AAV9)载体对小鼠肌肉的短期转导效果,初步寻找治疗杜氏肌营养不良症(DMD)安全有效的基因转导方式。方法使用AAV9包装携带荧光素酶报告基因的质粒构建AAV9-Luc病毒载体,经尾静脉注射和肌肉注射两种方式,分别向雌性BALB/c小鼠体内注射不同剂量的AAV9-Luc,3 d后于生物荧光成像仪下观察小鼠体内荧光表达情况,进而分析病毒在小鼠体内的分布情况。结果经尾静脉注射时,病毒在小鼠体内分布较为均匀广泛;而经肌肉注射时,除在注射部位有表达外,其他肌肉组织也有少量表达,但注射部位表达强度最高。此外,病毒在体内的分布随剂量增高而愈广泛,表达也随剂量增高而愈强烈。当肌肉注射剂量为1.26×10^(13)μg/kg时,病毒仅在注射部位附近及四肢、膈肌等肌肉组织中有表达,其他脏器表达不明显。结论在AAV9载体介导基因治疗DMD的技术中,低剂量(1.70×10^(11)μg/只,1.26×10^(13)μg/kg)肌肉注射是较为合适的转导方式。Objective To observe the short-term transduction effect of adeno-associated virus serotype 9(AAV9)vector on mouse muscle under different injection methods and doses,and to find a safe and effective gene transduction method for treating Duchenne muscular dystrophy(DMD).Methods The AAV9-Luc vector was constructed by AAV9 packaging plasmid carrying luciferase reporter gene.Female BALB/c mice were injected with different doses of AAV9-Luc by tail vein injection and intramuscular injection,respectively.Three days later,the fluorescence expression in mice was observed under bioluminescence imager,and the distribution of the virus in mice was analyzed.Results The virus was distributed evenly and widely in mice after tail vein injection.During intramuscular injection,a small amount of expression was also found in other muscle tissues except at the injection site,but the expression intensity was the highest at the injection site.In addition,the distribution and expression of the virus in vivo increased with the increase of the dose.When the intramuscular injection dose was 1.26×10^(13)μg/kg,the virus was only expressed near the injection site and in muscle tissues such as extremities and diaphragms,but not in other organs.Conclusion In the technique of AAV9 vector mediated gene therapy for DMD,intramuscular injection with a low dose(1.70×10^(11)μg/mouse,1.26×10^(13)μg/kg)is a suitable transduction method.

关 键 词：杜氏肌营养不良症 腺相关病毒9型 基因治疗 生物分布 肌肉 

分 类 号：R450[医药卫生—治疗学]