Unfolded Protein Response (UPR) During CHO Cell Production Culture

Du, Zhimei; Treiber, David; McCoy, Rebecca; He, Feng; Miller, Amanda; Han, Mei; Heath, Carole A.; Reddy, Pranhitha

doi:10.1021/bk-2013-1125.ch002

“…Cells activate the unfolded protein response (UPR) as an adaptive mechanism when the protein folding machinery of the ER is overwhelmed by the buildup of misfolded or unfolded proteins in the organelle 55,56 . Notably, overexpression of secreted and membrane-bound proteins, including antibodies 57 , GPCRs 58 , transporters 59 , and microbial opsins 60 , have been shown to trigger the UPR in cells. Accordingly, we sought to determine whether the UPR is induced by heterologous expression of Aqp1, a channel protein that must fold and mature inside the ER prior to reaching the cell membrane.…”

Section: Resultsmentioning

confidence: 99%

Engineering water exchange is a safe and effective method for magnetic resonance imaging in diverse cell types

Miller,

Chowdhury,

Hanson

et al. 2023

Preprint

0

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Aquaporin-1 (Aqp1), a water channel, has garnered significant interest for cell-based medicine and in vivo synthetic biology due to its ability to be genetically encoded to produce magnetic resonance signals by increasing the rate of water diffusion in cells. However, concerns regarding the effects of Aqp1 overexpression and increased membrane diffusivity on cell physiology have limited its widespread use as a deep-tissue reporter. In this study, we present evidence that Aqp1 generates strong diffusion-based magnetic resonance signals without adversely affecting cell viability or morphology in diverse cell lines derived from mice and humans. Our findings indicate that Aqp1 overexpression does not induce ER stress, which is frequently associated with heterologous expression of membrane proteins. Furthermore, we observed that Aqp1 expression had no detrimental effects on native biological activities, such as phagocytosis, immune response, insulin secretion, and tumor cell migration in the analyzed cell lines. These findings should serve to alleviate any lingering safety concerns regarding the utilization of Aqp1 as a genetic reporter and should foster its broader application as a noninvasive reporter for in vivo studies.

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Protein overproduction alters exosome secretion in Chinese hamster ovary cells

Steć,

Targońska,

Karkosińska

et al. 2023

Anal Bioanal Chem

3

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Despite the abundance of available cell lines, nearly 70% of all recombinant therapeutic proteins today are produced in Chinese hamster ovary (CHO) cells. The impact of protein overproduction on the secretion of exosomes by CHO cells has been investigated here. Increased secretion of extracellular vesicles (EVs) by protein overexpressing CHO cells was demonstrated with protein content assay, nanoparticle tracking analysis, and capillary electrophoresis. Our results revealed that a protein overproduction might induce EVs secretion, which might be accompanied by the sequestration and loading of overexpressed proteins into the exosomes. These findings are of vital importance for the manufacturing of therapeutics in CHO expression systems due to the risk of product loss during downstream processing of culture medium as well as the application of exosomes as nanocarriers of therapeutic proteins. The study indicates also the importance of culturing process control.

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Engineering water exchange is a safe and effective method for magnetic resonance imaging in diverse cell types

Miller,

Chowdhury,

Hanson

et al. 2024

J Biol Eng

1

0

View full text Add to dashboard Cite

Aquaporin-1 (Aqp1), a water channel, has garnered significant interest for cell-based medicine and in vivo synthetic biology due to its ability to be genetically encoded to produce magnetic resonance signals by increasing the rate of water diffusion in cells. However, concerns regarding the effects of Aqp1 overexpression and increased membrane diffusivity on cell physiology have limited its widespread use as a deep-tissue reporter. In this study, we present evidence that Aqp1 generates strong diffusion-based magnetic resonance signals without adversely affecting cell viability or morphology in diverse cell lines derived from mice and humans. Our findings indicate that Aqp1 overexpression does not induce ER stress, which is frequently associated with heterologous expression of membrane proteins. Furthermore, we observed that Aqp1 expression had no detrimental effects on native biological activities, such as phagocytosis, immune response, insulin secretion, and tumor cell migration in the analyzed cell lines. These findings should serve to alleviate any lingering safety concerns regarding the utilization of Aqp1 as a genetic reporter and should foster its broader application as a noninvasive reporter for in vivo studies.

show abstract

Unfolded Protein Response (UPR) During CHO Cell Production Culture

Cited by 3 publications

References 10 publications

Engineering water exchange is a safe and effective method for magnetic resonance imaging in diverse cell types

Engineering water exchange is a safe and effective method for magnetic resonance imaging in diverse cell types

Protein overproduction alters exosome secretion in Chinese hamster ovary cells

Engineering water exchange is a safe and effective method for magnetic resonance imaging in diverse cell types

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