Abstract
Background
Chinese hamster ovary (CHO) cells are the most predominantly utilized host for the production of monoclonal antibodies (mAbs) and other complex glycoproteins. A major challenge in the process of CHO cell culture is the occurrence of cell death following different stressful conditions, which hinders the production yield. Engineering genes involved in pathways related to cell death is a remarkable strategy to delay apoptosis, improve cell viability and enhance productivity. SIRT6 is a stress-responsive protein that regulates DNA repair, maintains genome integrity, and is critical for longevity and cell survival in organisms.
Methods and results
In this study, SIRT6 was stably overexpressed in CHO-K1 cells and the impact of its expression on apoptosis related gene expression profile, viability, apoptosis, and mAb productivity was investigated. While a significant increase was observed in Bcl-2 mRNA level, caspase-3 and Bax mRNA levels were decreased in the SIRT6 engineered cells compared to the parental CHO-K1 cells. Moreover, improved cell viability and decreased rate of apoptotic progression was observed in a SIRT6-derived clone in comparision to the CHO-K1 cells during 5 days of batch culture. anti-CD52 IgG1 mAb titers were improved up to 1.7- and 2.8-fold in SIRT6-derived clone during transient and stable expression, respectively.
Conclusions
This study indicates the positive effects of SIRT6 overexpression on cell viability and anti-CD52 IgG1 mAb expression in CHO-K1 cells. Further studies are needed to examine the potential of SIRT6-engineered host cells for the production of recombinant biotherapeutics in industrial settings.
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Data Availability
The datasets generated during and/or analysed during the current study are available from the corresponding author on reasonable request.
Abbreviations
- CHO:
-
Chinese hamster ovary
- mAb:
-
monoclonal antibody
- Bcl-2:
-
B-cell lymphoma 2
- Bax:
-
BCL2-Associated X
- BAK1:
-
BCL2 Antagonist/Killer 1
- Bcl-xL:
-
B-cell lymphoma-extra large
- PARP-1:
-
poly [ADP-ribose] polymerase 1
- Gcn5:
-
general control non-derepressible 5
- PGC-1α:
-
Peroxisome proliferator-activated receptor-gamma coactivator
- qRT-PCR:
-
quantitative real‑time PCR
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Acknowledgements
Authors wish to thank School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences for their support.
Funding
This study was supported by School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran (grants No. 15570 and 166).
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Experimental studies, data collection and data analysis were performed by NH. SHT and FS collaborated in experimental studies. Study conception, design, and supervision were performed by BK and AR. MR and JR contributed in data analysis and interpretation. The first draft of the manuscript was written by NH, and all authors commented on the previous drafts of the manuscript. All authors read and approved the final manuscript.
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Hashemi, N., Tabatabaee, S.H., Shams, F. et al. Overexpression of SIRT6 alleviates apoptosis and enhances cell viability and monoclonal antibody expression in CHO-K1 cells. Mol Biol Rep 50, 6019–6027 (2023). https://doi.org/10.1007/s11033-023-08483-5
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DOI: https://doi.org/10.1007/s11033-023-08483-5