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 110936

Subject 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

More information

Latest update

6/27/2022