针对线粒体复合体Ⅰ基因突变的Leigh综合征细胞模型的基因治疗研究  

The Study of Gene Therapy for a Cell Model of Leigh Syndrome with Mitochondrial Complex I Gene Mutation

作　　者：沈露茜 黄钰婷 方合志[3] 申钰琪 姜庆友 熊舒婷 李红智[3] SHEN Luxi;HUANG Yuting;FANG Hezhi;SHEN Yuqi;JIANG Qingyou;XIONG Shuting;LI Hongzhi(Department of Internal Neurology,Beijing Friendship Hospital,Capital Medical University,Beijing 100050,China;Department of Clinical Laboratory,Zhuji Affiliated Hospital of Wenzhou Medical University,Shaoxing 311800,China;Key Laboratory of Laboratory Medicine,Ministry of Education,School of Laboratory Medicine and Life Sciences,Wenzhou Medical University,Wenzhou 325035,China)

机构地区：[1]首都医科大学附属北京友谊医院神经内科,北京100050 [2]温州医科大学附属诸暨医院医学检验科,绍兴311800 [3]温州医科大学检验医学与生命科学学院,教育部检验医学重点实验室,温州325035

出　　处：《中国细胞生物学学报》2023年第5期753-761,共9页Chinese Journal of Cell Biology

基　　金：国家自然科学基金(批准号:81971291)资助的课题。

摘　　要：该研究旨在探讨转导酵母NDI1基因对线粒体ND1基因突变的Leigh综合征细胞模型的恢复效果,从而为线粒体复合体I基因突变所致Leigh综合征的基因治疗提供研究基础。已知线粒体复合体Ⅰ的ND1基因的m.3697G>A突变是Leigh综合征的致病突变之一。该研究采用已构建的携带该ND1基因突变的胞质杂合细胞作为线粒体复合体I基因突变的Leigh综合征细胞模型,将酵母NDI1基因的重组慢病毒转导至该细胞模型中表达NDI1蛋白(即酵母复合体I),检测NDI1蛋白对线粒体复合体I各方面功能的恢复效果。酵母NDI1基因转导该细胞模型后能高效表达并定位于线粒体。转导酵母NDI1基因可以恢复复合体I酶活性(外源酵母复合体Ⅰ的补偿)、线粒体有关的氧耗水平、线粒体偶联效率、线粒体有关的ATP水平,并且可以降低线粒体氧化应激水平、线粒体自噬水平。在线粒体复合体Ⅰ基因突变的Leigh综合征细胞模型中,酵母复合体Ⅰ可以替代性补偿线粒体的氧化磷酸化功能,并且可以缓解线粒体的氧化应激和自噬状态。该研究结果可以为线粒体复合体Ⅰ基因突变所致Leigh综合征的基因治疗提供研究基础。To provide a research basis for gene therapy of Leigh syndrome caused by mitochondrial complex I gene mutation,this study explored the therapeutic effect of transducing yeast NDI1 gene on the mitochondrial ND1 gene mutation in a cell model of Leigh syndrome.It is known that m.3697G>A mutation in ND1 gene of mitochondrial complex I is one of the pathogenic mutations of Leigh syndrome.In this study,the established cybrids carrying the ND1 gene mutation were used as the cell model of Leigh syndrome with mitochondrial complex I gene mutation.The recombinant lentivirus containing the yeast NDI1 gene was transduced into the cell model.The rescued effect of expressed NDI1 protein(namely yeast complex I)on all aspects of mitochondrial complex I functions were examined.NDI1 protein was highly expressed and localized in the mitochondria after NDI1 gene being transduced into the cell model.Transduction of NDI1 gene restored complex I enzyme activity(compensation by the exogenous yeast complex I),mitochondria-related oxygen consumption level,mitochondrial coupling efficiency,and mitochondria-related ATP level,and reduced mitochondrial oxidative stress and mitophagy level.In the cell model of Leigh syndrome with mitochondrial complex I gene mutation,yeast complex I can compensate the defective oxidative phosphorylation of mitochondria,and relieve mitochondrial oxidative stress and autophagic state.The results of this study may provide a basis for the gene therapy of Leigh syndrome caused by mutations in the mitochondrial complex I genes.

关 键 词：LEIGH综合征 线粒体复合体Ⅰ 基因突变 基因治疗 

分 类 号：R450[医药卫生—治疗学] R741[医药卫生—临床医学]