2. Academic Validation
  3. Apilimod enhances specific productivity in recombinant CHO cells through cell cycle arrest and mediation of autophagy

Apilimod enhances specific productivity in recombinant CHO cells through cell cycle arrest and mediation of autophagy

  • Biotechnol J. 2022 Dec 7;e2200147. doi: 10.1002/biot.202200147.
Jiang-Tao Lu 1 2 3,  Meng-Ke Xiao 2 3 4,  Ying-Ying Feng 1 2 3,  Xiao-Yin Wang 2 3 5,  Le-Le Qiu 5,  Yu-Rong Chai 6,  Tian-Yun Wang 2 3 5,  Yan-Long Jia 1 2 3
Affiliations

Affiliations

  • 1 School of Pharmacy, Xinxiang Medical University, Xinxiang, Henan, China.
  • 2 International Joint Research Laboratory for Recombinant Pharmaceutical Protein Expression System of Henan, Xinxiang Medical University, Xinxiang, Henan, China.
  • 3 Henan Engineering Research Center for Biopharmaceutical Innovation, Xinxiang Medical University, Xinxiang, Henan, China.
  • 4 School of Public Health, Xinxiang Medical University, Xinxiang, Henan, China.
  • 5 School of Basic Medicine, Xinxiang Medical University, Xinxiang, Henan, China.
  • 6 Department of Histology and Embryology, School of Basic Medical Sciences, Zhengzhou University, Zhengzhou, Henan, China.
PMID: 36478399 DOI: 10.1002/biot.202200147
Abstract

Chinese hamster ovary (CHO) cells are expected to acquire the ability to produce higher recombinant therapeutic protein levels using various strategies. Genetic engineering targeting the cell cycle and Autophagy pathways in the regulation of cell death in CHO cell cultures has received attention for enhancing the production of therapeutic proteins. In this study, we examined the small-molecule compound apilimod, which was found to have a positive influence on recombinant protein expression in CHO cells. This was confirmed by selective blocking of the cell cycle at the G0/G1 phase. Apilimod treatment resulted in decreased expression of cyclin-dependent kinase 3 (CDK3) and Cyclin C and increased expression of cyclin-dependent kinase suppressor p27Kip1, which are critical regulators of G1 cell cycle progression and important targets controlling cell proliferation. Furthermore, total transcription factor EB (TFEB) was lower in apilimod-treated CHO cells than in control cells, resulting in decreased lysosome biogenesis and Autophagy with apilimod treatment. These multiple effects demonstrate the potential of apilimod for development as a novel enhancer for the production of recombinant proteins in CHO cell engineering.

Keywords

Chinese hamster ovary (CHO) cells, therapeutic protein expression; apilimod, autophagy, cell cycle.

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