Comparative transcriptional analysis of mouse hybridoma and recombinant Chinese hamster ovary cells undergoing butyrate treatment

Gatti, Marcela De Leon; Wlaschin, Katie F.; Nissom, Peter Morin; Yap, Melvin J.; Hu, Wei Shou

doi:10.1263/jbb.103.82

Cited by 79 publications

(67 citation statements)

References 26 publications

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“…Biological processes identified to be overrepresented included mainly lipid biosynthetic processes and negative regulation of apoptosis. This independently confirms observations in earlier studies focussing on transcriptomic profiling of individual CHO cell lines with a highqP phenotype that identified ER, Golgi and intracellular transport processes, which are all membrane-associated, as contributing to productivity [7,19,20,50].…”

Section: Regression Of Changes In Gene Expression To Changes In Qpsupporting

confidence: 90%

“…In the case of the proteasome, we only find Psmd3 (Rpn3), a structural component of the lid of the 26S proteasome, significantly changing (q<0.05) with increasing qP. Such a general trend towards down-regulation of the structural genes of the proteasome [50,60,61] or related ubiquitin ligases [7] has also been found in previous studies, and was also identified in recombinant yeast [62]. In combination with the results for the ER it seems that, although the ER is upregulated for high protein cargo, in the high producing subclones this is not happening under stress response.…”

Section: Proteasomementioning

confidence: 61%

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Microarray profiling of preselected CHO host cell subclones identifies gene expression patterns associated with in‐creased production capacity

Harreither

Hackl

Pichler

et al. 2015

Biotechnology Journal

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Over the last three decades, product yields from CHO cells have increased dramatically, yet specific productivity (qP) remains a limiting factor. In a previous study, using repeated cell-sorting, we have established different host cell subclones that show superior transient qP over their respective parental cell lines (CHO-K1, CHO-S). The transcriptome of the resulting six cell lines in different biological states (untransfected, mock transfected, plasmid transfected) was first explored by hierarchical clustering and indicated that gene activity associated with increased qP did not stem from a certain cellular state but seemed to be inherent for a high qP host line. We then performed a novel gene regression analysis identifying drivers for an increase in qP. Genes significantly implicated were first systematically tested for enrichment of GO terms using a Bayesian approach incorporating the hierarchical structure of the GO term tree. Results indicated that specific cellular components such as nucleus, ER, and Golgi are relevant for cellular productivity. This was complemented by targeted GSA that tested functionally homogeneous, manually curated subsets of KEGG pathways known to be involved in transcription, translation, and protein processing. Significantly implicated pathways included mRNA surveillance, proteasome, protein processing in the ER and SNARE interactions in vesicular transport.

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Section: Regression Of Changes In Gene Expression To Changes In Qpsupporting

confidence: 90%

Section: Proteasomementioning

confidence: 61%

Microarray profiling of preselected CHO host cell subclones identifies gene expression patterns associated with in‐creased production capacity

Harreither

Hackl

Pichler

et al. 2015

Biotechnology Journal

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“…Acetylation of histones facilitates the relaxation of the chromatin and consequently, transcription factors can bind and alter gene expression of various cellular processes (Riggs et al 1977). Butyrate selectively increases transcription of genes, and the changes in gene expression due to butyrate treatment in CHO cells have been studied at the transcriptomic and proteomic levels (Yee et al 2008;De Leon et al 2007). Butyrate treatment in CHO cells affects cellular processes such as cytoskeleton reorganization, protein transport, nucleosome assembly, nucleotide metabolism, glycosylation, protein folding, oxidative stress responses, lipid metabolism, cholesterol biosynthesis, glycolysis, cell cycle progression, and apoptosis (Yee et al 2008;De Leon et al 2007).…”

Section: Butyrate Work As a Histone Deacetylase Inhibitor Andmentioning

confidence: 99%

“…Butyrate treatment in CHO cells affects cellular processes such as cytoskeleton reorganization, protein transport, nucleosome assembly, nucleotide metabolism, glycosylation, protein folding, oxidative stress responses, lipid metabolism, cholesterol biosynthesis, glycolysis, cell cycle progression, and apoptosis (Yee et al 2008;De Leon et al 2007). Butyrate has been used in cell engineering strategies, and butyrate treatment in CHO cells often increases recombinant protein expression (Chun et al 2003;De Leon et al 2007;Hendrick et al 2001;Kim and Lee 2000;Chotigeat et al 1994;Mimura et al 2001;Wang et al 2002;Jiang and Sharfstein 2008). The current work explores the effect of butyrate on glycosaminoglycan (GAG) profiles in recombinant CHO cells in our efforts to produce a bioengineered heparin (HP).…”

Section: Butyrate Work As a Histone Deacetylase Inhibitor Andmentioning

confidence: 99%

“…Sodium butyrate (CH 3 CH 2 CH 2 CO 2 -) is a histone deacetylase inhibitor that modulates gene expression of cellular processes, particularly transgene expression in Chinese hamster ovary (CHO) cells (Chotigeat et al 1994;Chun et al 2003;De Leon et al 2007;Hendrick et al 2001;Jiang and Sharfstein 2008;Kim and Lee 2000;Mimura et al 2001;Wang et al 2002).…”

Section: Introductionmentioning

confidence: 99%

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Modulation of heparan sulfate biosynthesis by sodium butyrate in recombinant CHO cells

et al. 2014

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Sodium butyrate, a histone deacetylase inhibitor, has been used to improve transgene expression in Chinese hamster ovary (CHO) cells. The current study explores the impact of butyrate treatment on heparan sulfate (HS) biosynthesis and structural composition in a recombinant CHO-S cell line expressing enzymes in the heparin (HP)/(HS) biosynthetic pathway (Dual-10 stably expressing NDST2 and HS3st1). Flow cytometric analysis showed that antithrombin binding was increased in Dual-10 cells and basic fibroblast growth factor binding was decreased in response to sodium butyrate treatment. The results were in agreement with the AMAC-LCMS (2-aminoacridine-tagged HS/HP analysis by liquid chromatography mass spectrometry) data that showed that there was an increase in heparan sulfate tri-sulfated disaccharides and a decrease in N-sulfated disaccharides in the butyrate-treated cells. However, we could not detect any changes in the chondroitin sulfate pathway in Dual-10 cells treated with butyrate. The current study is the first to report the effect of butyrate on glycosaminoglycan profiles.

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Animal Cells, Hybridomas, Human Antibody Production

Warren¹,

Subramanian²

2010

Encyclopedia of Industrial Biotechnology

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Cellular response to perturbations in the microenvironment manifests as a surge of physiological changes that have great impact on cellular metabolism and propagation. In bioprocess systems, mammalian cells engineered for the production of monoclonal antibodies or therapeutic proteins are sensitive to fluctuations in the culture environment. These fluctuations often directly translate to changes in maximum attainable cell concentration and/or specific protein productivity. In conjunction with feeding strategy optimization, the ability to understand and optimize the physical environment of animal culture systems is key to achieving a process that delivers consistent production of high titer batches. In this article we have reviewed comprehensively two critical physical culture parameters: osmolality and temperature, for their effects on general cellular physiology as well as their impacts on overall culture performance in bioprocess systems. Extreme levels of osmolality lead to volume‐regulatory responses, cytoskeletal reorganization, and trigger distinct cellular signaling pathways. Cellular responses to hypothermic conditions include reduced metabolic and growth rates, reduction of apoptosis frequency, and a cold‐shock response which involves the increased synthesis of several proteins. Exposure of cells to hyperosmotic conditions and/or subphysiological temperature (32–35°C) has been shown to enhance specific protein productivity in bioprocess systems. As live viruses are also commonly produced in industrial cell culture systems for vaccine processes, we have extended the scope of this article to include the effects of fluctuations in culture osmolality and temperature on the production of live viruses.

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Comparative transcriptional analysis of mouse hybridoma and recombinant Chinese hamster ovary cells undergoing butyrate treatment

Cited by 79 publications

References 26 publications

Microarray profiling of preselected CHO host cell subclones identifies gene expression patterns associated with in‐creased production capacity

Microarray profiling of preselected CHO host cell subclones identifies gene expression patterns associated with in‐creased production capacity

Modulation of heparan sulfate biosynthesis by sodium butyrate in recombinant CHO cells

Animal Cells, Hybridomas, Human Antibody Production

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