Enhanced metabolism and negative regulation of ER stress support higher erythropoietin production in HEK293 cells
Artikel i vetenskaplig tidskrift, 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
Författare
Rasool Saghaleyni
Chalmers, Biologi och bioteknik, Systembiologi
Magdalena Malm
Kungliga Tekniska Högskolan (KTH)
Noah Moruzzi
Karolinska Institutet
Jan Zrimec
Chalmers, Biologi och bioteknik, Systembiologi
Ronia Razavi
Kungliga Tekniska Högskolan (KTH)
Num Wistbacka
Kungliga Tekniska Högskolan (KTH)
Hannes Thorell
Kungliga Tekniska Högskolan (KTH)
Anton Pintar
Kungliga Tekniska Högskolan (KTH)
Andreas Hober
Kungliga Tekniska Högskolan (KTH)
F. Edfors
Kungliga Tekniska Högskolan (KTH)
Veronique Chotteau
Kungliga Tekniska Högskolan (KTH)
Per Olof Berggren
Karolinska Institutet
Luigi Grassi
AstraZeneca AB
Aleksej Zelezniak
Chalmers, Biologi och bioteknik, Systembiologi
Thomas Svensson
Chalmers, Biologi och bioteknik, Systembiologi
Diane Hatton
AstraZeneca AB
Jens B Nielsen
Chalmers, Biologi och bioteknik, Systembiologi
Danmarks Tekniske Universitet (DTU)
Jonathan Robinson
Chalmers, Biologi och bioteknik, Systembiologi
J. Rockberg
Kungliga Tekniska Högskolan (KTH)
Cell Reports
22111247 (eISSN)
Vol. 39 11 110936Ämneskategorier
Biokemi och molekylärbiologi
Mikrobiologi
Medicinsk bioteknologi (med inriktning mot cellbiologi (inklusive stamcellsbiologi), molekylärbiologi, mikrobiologi, biokemi eller biofarmaci)
DOI
10.1016/j.celrep.2022.110936
PubMed
35705050