Overexpression of GADD34 Enhances Production of Recombinant Human Antithrombin III in Chinese Hamster Ovary Cells

Ômasa, Takeshi; Takami, Takashi; Ohya, Takeshi; Kiyama, Eriko; Hayashi, Tetsuji; Nishii, Hiroko; Miki, Hajime; Kobayashi, Kaoru; Honda, Kohsuke; Ohtake, Hisao

doi:10.1263/jbb.106.568

Cited by 55 publications

(24 citation statements)

References 22 publications

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“…In order to obtain productive cells with higher specific production rates, it is important to engineer the secretory pathway (Hou et al 2012;Le Fourn et al 2013). ATF4 overexpression can enhance the dephosphorylation of phosphorylated eIF2a by activating the expression of GADD34 Omasa et al 2008). In addition, the production of heterologous protein is always accompanied by overproduction of disulfide bonds inside the ER, which leads to the production of reactive oxygen species (ROS) .…”

Section: Resultsmentioning

confidence: 99%

Improved antibody production in Chinese hamster ovary cells by ATF4 overexpression

et al. 2013

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To improve antibody production in Chinese hamster ovary (CHO) cells, the humanized antibody-producing CHO DP-12-SF cell line was transfected with the gene encoding activating transcription factor 4 (ATF4), a central factor in the unfolded protein response. Overexpression of ATF4 significantly enhanced the production of antibody in the CHO DP-12-SF cell line. The specific IgG production rate of in the ATF4-overexpressing CHO-ATF4-16 cells was approximately 2.4 times that of the parental host cell line. Clone CHO-ATF4-16 did not show any change in growth rate compared with the parental cells or mock-transfected CHO-DP12-SF cells. The expression levels of mRNAs encoding both the antibody heavy and light chains in the CHO-ATF4-16 clone were analyzed. This analysis showed that ATF4 overexpression improved the total production and specific production rate of antibody without affecting the mRNA transcription level. These results indicate that ATF4 overexpression is a promising method for improving recombinant IgG production in CHO cells.

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Section: Resultsmentioning

confidence: 99%

Improved antibody production in Chinese hamster ovary cells by ATF4 overexpression

et al. 2013

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“…Accumulation of misfolded protein within the ER compartment triggers the unfolded protein and ER overload responses and thereby causes an undesirable decline in cell specific productivity. Coexpression of the UPR-associated transcription factor GADD34 (gene encoding growth arrest and DNA damage inducible protein 34) has been shown to increase production of recombinant human antithrombin III from CHO cells [84]. A very interesting approach to overcome bottlenecks in intracellular protein processing is based on artificial expansion of the ER capacity.…”

Section: Engineering Approaches Aimed At Improving Protein Secretion mentioning

confidence: 99%

3.5 Manipulation of Cell Growth, Metabolism and Product Quality Attributes

Horsten¹,

Winkler²,

Sandig³

2014

Animal Cell Biotechnology

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“…In some experiments, pcDNA3.CD2-heIF2a-S51A encoding a dominant-negative mutant of eIF2a (eIF2a-DN) (provided by Dr David Ron, University of Cambridge) 29 or pcDNA3.1-GADD34 coding for growth arrest and DNA damage gene 34 (GADD34) (provided by Dr Takeshi Omasa, Osaka University) 30 was co-transfected to evaluate involvement of eIF2a. pGL2-F-1005/ þ 93Smad1 that introduces a luciferase gene under the control of the Smad1 gene promoter (provided by Dr Yuewen Gong, University of Manitoba) 31 was also used to evaluate the effect of 3 0 -deoxyadenosine.…”

Section: Reporter Assaymentioning

confidence: 99%

Blockade of Smad signaling by 3′-deoxyadenosine: a mechanism for its anti-fibrotic potential

Johno

Nakajima

et al. 2013

Laboratory Investigation

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Cordyceps militaris has been used in Eastern countries for the treatment of various diseases including chronic kidney diseases. However, there are no reports that identified its active entities and molecular mechanisms underlying its therapeutic effectiveness. 3 0 -Deoxyadenosine is a major nucleoside derivative isolated from C. militaris. Some reports suggested that both C. militaris and 3 0 -deoxyadenosine have anti-inflammatory and anti-fibrotic effects. In the present report, we investigated whether and how 3 0 -deoxyadenosine interferes with fibrogenic processes in the kidney. For this purpose, we examined effects of 3 0 -deoxyadenosine on the expression of collagens triggered by transforming growth factor-b (TGF-b1) and bone morphogenetic protein-4 (BMP-4), especially focusing on the regulation of Smad signaling in vitro and in vivo. We found that 3 0 -deoxyadenosine suppressed expression of collagens induced by TGF-b1 and BMP-4 dose dependently. This suppression occurred at the transcriptional level and was correlated with blunted activation of the CAGA box and the BMP-responsive element. The suppressive effect on the TGF-b/BMP signaling was mediated mainly by adenosine transporter and partially by the A3 adenosine receptor, but not A1/A2 adenosine receptors. 3 0 -Deoxyadenosine reduced levels of both phosphorylated and total Smad proteins (Smad1, 2 and 3) dose dependently. It was mainly ascribed to transcriptional suppression, but not to enhanced protein degradation and eIF2a-mediated translational suppression. Consistent with the in vitro results, in vivo administration with 3 0 -deoxyadenosine reduced the levels of phosphorylated and total Smad proteins, as well as the levels of Smad mRNAs, in the kidney subjected to unilateral ureteral obstruction. It was associated with blunted induction of type I collagen and a-smooth muscle actin, a decrease in the number of interstitial myofibroblasts and reduced fibrotic area. These results suggest that 3 0 -deoxyadenosine interferes with the TGF-b and BMP signaling via downregulation of Smads, which may underlie the anti-fibrotic effect of this agent. 3 0 -Deoxyadenosine may be useful for therapeutic intervention in various TGF-b-related fibrotic disorders.

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Overexpression of GADD34 Enhances Production of Recombinant Human Antithrombin III in Chinese Hamster Ovary Cells

Cited by 55 publications

References 22 publications

Improved antibody production in Chinese hamster ovary cells by ATF4 overexpression

Improved antibody production in Chinese hamster ovary cells by ATF4 overexpression

3.5 Manipulation of Cell Growth, Metabolism and Product Quality Attributes

Blockade of Smad signaling by 3′-deoxyadenosine: a mechanism for its anti-fibrotic potential

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