Enhanced metabolism and negative regulation of ER stress support higher erythropoietin production in HEK293 cells
Journal article, 2022
Recombinant protein production can cause severe stress on cellular metabolism, resulting in limited titer and product quality. To investigate cellular and metabolic characteristics associated with these limitations, we compare HEK293 clones producing either erythropoietin (EPO) (secretory) or GFP (non-secretory) protein at different rates. Transcriptomic and functional analyses indicate significantly higher metabolism and oxidative phosphorylation in EPO producers compared with parental and GFP cells. In addition, ribosomal genes exhibit specific expression patterns depending on the recombinant protein and the production rate. In a clone displaying a dramatically increased EPO secretion, we detect higher gene expression related to negative regulation of endoplasmic reticulum (ER) stress, including upregulation of ATF6B, which aids EPO production in a subset of clones by overexpression or small interfering RNA (siRNA) knockdown. Our results offer potential target pathways and genes for further development of the secretory power in mammalian cell factories.
GFP
HEK293
ATF6B
protein production
CP: Molecular biology
erythropoietin
ribosome heterogeneity
CP: Cell biology
secretory pathways
Cell engineering
Author
Rasool Saghaleyni
Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology
Magdalena Malm
Royal Institute of Technology (KTH)
Noah Moruzzi
Karolinska Institutet
Jan Zrimec
Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology
Ronia Razavi
Royal Institute of Technology (KTH)
Num Wistbacka
Royal Institute of Technology (KTH)
Hannes Thorell
Royal Institute of Technology (KTH)
Anton Pintar
Royal Institute of Technology (KTH)
Andreas Hober
Royal Institute of Technology (KTH)
F. Edfors
Royal Institute of Technology (KTH)
Veronique Chotteau
Royal Institute of Technology (KTH)
Per Olof Berggren
Karolinska Institutet
Luigi Grassi
AstraZeneca AB
Aleksej Zelezniak
Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology
Thomas Svensson
Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology
Diane Hatton
AstraZeneca AB
Jens B Nielsen
Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology
Technical University of Denmark (DTU)
Jonathan Robinson
Chalmers, Biology and Biological Engineering, Systems and Synthetic Biology
J. Rockberg
Royal Institute of Technology (KTH)
Cell Reports
22111247 (eISSN)
Vol. 39 11 110936Subject Categories
Biochemistry and Molecular Biology
Microbiology
Medical Biotechnology (with a focus on Cell Biology (including Stem Cell Biology), Molecular Biology, Microbiology, Biochemistry or Biopharmacy)
DOI
10.1016/j.celrep.2022.110936
PubMed
35705050