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Multiplex genome editing eliminates lactate production without impacting growth rate in mammalian cells

Hefzi, Hooman ; Martínez-Monge, Iván ; Marin de Mas, Igor ; Cowie, Nicholas Luke ; Toledo, Alejandro Gomez LU ; Noh, Soo Min ; Karottki, Karen Julie la Cour ; Decker, Marianne ; Arnsdorf, Johnny and Camacho-Zaragoza, Jose Manuel , et al. (2025) In Nature Metabolism
Abstract

The Warburg effect, which describes the fermentation of glucose to lactate even in the presence of oxygen, is ubiquitous in proliferative mammalian cells, including cancer cells, but poses challenges for biopharmaceutical production as lactate accumulation inhibits cell growth and protein production. Previous efforts to eliminate lactate production in cells for bioprocessing have failed as lactate dehydrogenase is essential for cell growth. Here, we effectively eliminate lactate production in Chinese hamster ovary and in the human embryonic kidney cell line HEK293 by simultaneous knockout of lactate dehydrogenases and pyruvate dehydrogenase kinases, thereby removing a negative feedback loop that typically inhibits pyruvate conversion to... (More)

The Warburg effect, which describes the fermentation of glucose to lactate even in the presence of oxygen, is ubiquitous in proliferative mammalian cells, including cancer cells, but poses challenges for biopharmaceutical production as lactate accumulation inhibits cell growth and protein production. Previous efforts to eliminate lactate production in cells for bioprocessing have failed as lactate dehydrogenase is essential for cell growth. Here, we effectively eliminate lactate production in Chinese hamster ovary and in the human embryonic kidney cell line HEK293 by simultaneous knockout of lactate dehydrogenases and pyruvate dehydrogenase kinases, thereby removing a negative feedback loop that typically inhibits pyruvate conversion to acetyl-CoA. These cells, which we refer to as Warburg-null cells, maintain wild-type growth rates while producing negligible lactate, show a compensatory increase in oxygen consumption, near total reliance on oxidative metabolism, and higher cell densities in fed-batch cell culture. Warburg-null cells remain amenable for production of diverse biotherapeutic proteins, reaching industrially relevant titres and maintaining product glycosylation. The ability to eliminate lactate production may be useful for biotherapeutic production and provides a tool for investigating a common metabolic phenomenon.

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publishing date
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Contribution to journal
publication status
epub
subject
in
Nature Metabolism
publisher
Springer Nature
external identifiers
  • scopus:85217518106
  • pmid:39809975
ISSN
2522-5812
DOI
10.1038/s42255-024-01193-7
language
English
LU publication?
yes
id
562157c4-62ed-485d-875a-c889657ac06d
date added to LUP
2025-01-16 08:41:48
date last changed
2025-07-05 20:01:54
@article{562157c4-62ed-485d-875a-c889657ac06d,
  abstract     = {{<p>The Warburg effect, which describes the fermentation of glucose to lactate even in the presence of oxygen, is ubiquitous in proliferative mammalian cells, including cancer cells, but poses challenges for biopharmaceutical production as lactate accumulation inhibits cell growth and protein production. Previous efforts to eliminate lactate production in cells for bioprocessing have failed as lactate dehydrogenase is essential for cell growth. Here, we effectively eliminate lactate production in Chinese hamster ovary and in the human embryonic kidney cell line HEK293 by simultaneous knockout of lactate dehydrogenases and pyruvate dehydrogenase kinases, thereby removing a negative feedback loop that typically inhibits pyruvate conversion to acetyl-CoA. These cells, which we refer to as Warburg-null cells, maintain wild-type growth rates while producing negligible lactate, show a compensatory increase in oxygen consumption, near total reliance on oxidative metabolism, and higher cell densities in fed-batch cell culture. Warburg-null cells remain amenable for production of diverse biotherapeutic proteins, reaching industrially relevant titres and maintaining product glycosylation. The ability to eliminate lactate production may be useful for biotherapeutic production and provides a tool for investigating a common metabolic phenomenon.</p>}},
  author       = {{Hefzi, Hooman and Martínez-Monge, Iván and Marin de Mas, Igor and Cowie, Nicholas Luke and Toledo, Alejandro Gomez and Noh, Soo Min and Karottki, Karen Julie la Cour and Decker, Marianne and Arnsdorf, Johnny and Camacho-Zaragoza, Jose Manuel and Kol, Stefan and Schoffelen, Sanne and Pristovšek, Nuša and Hansen, Anders Holmgaard and Miguez, Antonio A and Bjørn, Sara Petersen and Brøndum, Karen Kathrine and Javidi, Elham Maria and Jensen, Kristian Lund and Stangl, Laura and Kreidl, Emanuel and Kallehauge, Thomas Beuchert and Ley, Daniel and Ménard, Patrice and Petersen, Helle Munck and Sukhova, Zulfiya and Bauer, Anton and Casanova, Emilio and Barron, Niall and Malmström, Johan and Nielsen, Lars K and Lee, Gyun Min and Kildegaard, Helene Faustrup and Voldborg, Bjørn G and Lewis, Nathan E}},
  issn         = {{2522-5812}},
  language     = {{eng}},
  month        = {{01}},
  publisher    = {{Springer Nature}},
  series       = {{Nature Metabolism}},
  title        = {{Multiplex genome editing eliminates lactate production without impacting growth rate in mammalian cells}},
  url          = {{http://dx.doi.org/10.1038/s42255-024-01193-7}},
  doi          = {{10.1038/s42255-024-01193-7}},
  year         = {{2025}},
}