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 110936

Ämneskategorier

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

Mer information

Senast uppdaterat

2022-06-27