GAPDH suppresses adenovirus-induced oxidative stress and enables a superfast production of recombinant adenovirus  

作　　者：Guozhi Zhao Piao Zhao Yonghui Wang Hui Zhang Yi Zhu Jiamin Zhong Wulin You Guowei Shen Changi Luo Ou Mei Xingye Wu Jingjing Li Yi Shu Hongwei Wang Wiliam Wagstaff Hue HLuu Yang Bi Lewis LShi Russell RReid Tong-Chuan He Li Jiang Wei Tang Jiaming Fan Ziwei Tang 

机构地区：[1]Departments of Urology,Endocrinology,Orthopedic Surgery,and Gastroenterological Surgery,The First Affiliated Hospital of Chongqing Medical University,Chongqing 400016,China [2]Molecular Oncology Laboratory,Department of Orthopaedic Surgery and Rehabilitation Medicine,The University of Chicago Medical Center,Chicago,IL 60637,USA [3]Ministry of Education Key Laboratory of Diagnostic Medicine,and Department of Clinical 4Biochemistry,School of Laboratory Medicine,Chongqing Medical University,Chongqing 400016,China [4]Department of Geriatrics,Xinhua Hospital,Shanghai Jiao-Tong University School of Medicine,Shanghai 200000,China [5]The Breast Cancer Center,Chongqing University Cancer Hospital,Chongqing 4000430,China [6]Department of Orthopaedic Surgery,Beijing Hospital,Chinese Academy of Medical Sciences&Peking Union Medical College,Beijing 100730,China [7]Department of Orthopaedic Surgery,Wuxi Hospital Affiliated to Nanjing University of Chinese Medicine,Wuxi,Jiangsu 214071,China [8]Department of Orthopaedic Surgery,Beng Medical Center,The Affiliated Beng Hospital of Nanjing Medical University,Nanjing,Jiangsu 210019,China [9]Department of Orthopaedic Surgery,Yibin Second People's Hospital,Affiliated with West China School of Medicine,Yibin,Sichuan 644000,China [10]Department of Orthopedics,Jiangxi Hospital of Traditional Chinese Medicine,Jiangxi University of Traditional Chinese Medicine,Nanchang,Jiangxi 330006,China [11]Department of Oncology,The Affiliated Hospital of Weifang Medical University,Weifang,Shandong 261053,China [12]Stem Cell Biology and Therapy Laboratory of the Pediatric Research Institute,The National Clinical Research Center for Child Health and Disorders,and Ministry of Education Key Laboratory of Child Development and Disorders,the Children's Hospital of Chongqing Medical University,Chongqing 400016,China [13]Division of Research and Development,Decoding Therapeutics,Inc.,Mt Prospect,IL 60056,USA [14]Laboratory of Craniofacial Biology and Development,Department of Surgery Section of Plastic Surg

出　　处：《Genes & Diseases》2024年第6期367-380,共14页基因与疾病（英文）

基　　金：supported in part by research grants from the Natural Science Foundation of China (No.82000744 to ZT,and 82102696 to J.F.);the Chongqing Bayu Young Scholar Award (China) (to J.F.);the 2019 Funding for Postdoctoral Research (Chongqing Human Resources and Social Security Bureau of China) (No.298 to J.F.);the National Institutes of Health (No.CA226303 to T.C.H.,DE030480 to R.R.R.);supported by the Medical Scientist Training Program of the National Institutes of Health (USA) (No.T32 GM007281);supported in part by The University of Chicago Cancer Center Support Grant (No.P30CA014599);the National Center for Advancing Translational Sciences of the National Institutes of Health through grant number 2UL1TR002389-06 that funds the Institute for Translational Medicine (ITM);supported by the Mabel Green Myers Research Endowment Fund and The University of Chicago Orthopaedics Alumni Fund.

摘　　要：Recombinant adenovirus(rAdV)is a commonly used vector system for gene transfer.Efficient initial packaging and subsequent production of rAdV remains time-consuming and labor-intensive,possibly attributable to rAdv infection-associated oxidative stress and reactive oxygen species(ROS)production.Here,we show that exogenous GAPDH expression mitigates adenovirus-induced ROS-associated apoptosis in HEK293 cells,and expedites adenovirus production.By stably overexpressing GAPDH in HEK293(293G)and 293pTP(293GP)cells,respectively,we demonstrated that rAdV-induced RoS production and cell apoptosis were significantly suppressed in 293G and 293GP cells.Transfection of 293G cells with adenoviral plasmid pAd-G2Luc yielded much higher titers of Ad-G2Luc at day 7 than that in HEK293 cells.Similarly,Ad-G2Luc was amplified more efficiently in 293G than in HEK293 cells.We further showed that transfection of 293GP cells with pAd-G2Luc produced much higher titers of Ad-G2Luc at day 5 than that of 293pTP cells.293GP cells amplified the Ad-G2Luc much more efficiently than 293pTP cells,indicating that exogenous GAPDH can further augment pTP-enhanced adenovirus production.These results demonstrate that exogenous GAPDH can effectively suppress adenovirus-induced ROS and thus accelerate adenovirus production.Therefore,the engineered 293GP cells represent a superfast rAdV production system for adenovirus-based gene transfer and gene therapy.

关 键 词：GAPDH GENETHERAPY Oxidative stress Packaging cell line Reactive oxygen species Recombinant adenovirus 

分 类 号：R394[医药卫生—医学遗传学]