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Full Length Article | Open Access

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

Guozhi Zhaoa,b, Piao Zhaoa,b,c, Yonghui Wangb,d, Hui Zhangb,e, Yi Zhub,f, Jiamin Zhongb,c, Wulin Youb,g, Guowei Shenb,h, Changqi Luob,i, Ou Meib,j, Xingye Wua,b, Jingjing Lib,k, Yi Shub,m, Hongwei Wangn, William Wagstaffb, Hue H. Luub, Yang Bib,m, Lewis L. Shib, Russell R. Reidb,o, Tong-Chuan Heb,o, Li Jianga( ), Wei Tanga( ), Jiaming Fanb,c( ), Ziwei Tanga( )
Departments of Urology, Endocrinology, Orthopedic Surgery, and Gastroenterological Surgery, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, China
Molecular Oncology Laboratory, Department of Orthopaedic Surgery and Rehabilitation Medicine, The University of Chicago Medical Center, Chicago, IL 60637, USA
Ministry of Education Key Laboratory of Diagnostic Medicine, and Department of Clinical Biochemistry, School of Laboratory Medicine, Chongqing Medical University, Chongqing 400016, China
Department of Geriatrics, Xinhua Hospital, Shanghai Jiao-Tong University School of Medicine, Shanghai 200000, China
The Breast Cancer Center, Chongqing University Cancer Hospital, Chongqing 4000430, China
Department of Orthopaedic Surgery, Beijing Hospital, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing 100730, China
Department of Orthopaedic Surgery, Wuxi Hospital Affiliated to Nanjing University of Chinese Medicine, Wuxi, Jiangsu 214071, China
Department of Orthopaedic Surgery, Benq Medical Center, The Affiliated Benq Hospital of Nanjing Medical University, Nanjing, Jiangsu 210019, China
Department of Orthopaedic Surgery, Yibin Second People's Hospital, Affiliated with West China School of Medicine, Yibin, Sichuan 644000, China
Department of Orthopedics, Jiangxi Hospital of Traditional Chinese Medicine, Jiangxi University of Traditional Chinese Medicine, Nanchang, Jiangxi 330006, China
Department of Oncology, The Affiliated Hospital of Weifang Medical University, Weifang, Shandong 261053, China
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
Division of Research and Development, Decoding Therapeutics, Inc., Mt Prospect, IL 60056, USA
Laboratory of Craniofacial Biology and Development, Department of Surgery Section of Plastic Surgery, The University of Chicago Medical Center, Chicago, IL 60637, USA

Peer review under responsibility of Chongqing Medical University.

Show Author Information

Abstract

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.

Keywords

GAPDHGene therapyOxidative stressPackaging cell lineReactive oxygen speciesRecombinant adenovirus

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Genes & Diseases
Volume 11 Issue 6,
November 2024
Article number: 101344
DOI: 10.1016/j.gendis.2024.101344
Cite this article:
Zhao G, Zhao P, Wang Y, et al. GAPDH suppresses adenovirus-induced oxidative stress and enables a superfast production of recombinant adenovirus. Genes & Diseases, 2024, 11(6): 101344. https://doi.org/10.1016/j.gendis.2024.101344

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Received: 20 March 2024
Accepted: 15 May 2024
Published: 31 May 2024
© 2024 The Authors.

This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/).
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