Lessons from GNE-deficient embryonic stem cells: sialic acid biosynthesis is involved in proliferation and gene expression

Weidemann, Wenke; Klukas, Christian; Klein, Andreas; Simm, Andreas; Schreiber, Falk; Horstkorte, Rüdiger

doi:10.1093/glycob/cwp153

Cited by 35 publications

(35 citation statements)

References 53 publications

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“…5), suggested that IFN-g sialylation was bottlenecked at the CMP-SA synthesis stage. Furthermore, UAE expression decreases with time and is in agreement with findings from Weidemann et al (2009) who found that UAE expression is directly correlated with growth. These findings implied that during ''stationary'' non-growth phase of a typical process, when the majority of recombinant protein is produced, sialylation would be impaired due to low UAE expression.…”

Section: )supporting

confidence: 91%

Profiling ofN‐glycosylation gene expression in CHO cell fed‐batch cultures

Wong

Goh

et al. 2010

Biotech & Bioengineering

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One of the goals of recombinant glycoprotein production is to achieve consistent glycosylation. Although many studies have examined the changes in the glycosylation quality of recombinant protein with culture, very little has been done to examine the underlying changes in glycosylation gene expression as a culture progresses. In this study, the expression of 24 genes involved in N-glycosylation were examined using quantitative RT PCR to gain a better understanding of recombinant glycoprotein glycosylation during production processes. Profiling of the N-glycosylation genes as well as concurrent analysis of glycoprotein quality was performed across the exponential, stationary and death phases of a fed-batch culture of a CHO cell line producing recombinant human interferon-gamma (IFN-gamma). Of the 24 N-glycosylation genes examined, 21 showed significant up- or down-regulation of gene expression as the fed-batch culture progressed from exponential, stationary and death phase. As the fed-batch culture progressed, there was also an increase in less sialylated IFN-gamma glycoforms, leading to a 30% decrease in the molar ratio of sialic acid to recombinant IFN-gamma. This correlated with decreased expression of genes involved with CMP sialic acid synthesis coupled with increased expression of sialidases. Compared to batch culture, a low glutamine fed-batch strategy appears to need a 0.5 mM glutamine threshold to maintain similar N-glycosylation genes expression levels and to achieve comparable glycoprotein quality. This study demonstrates the use of quantitative real time PCR method to identify possible "bottlenecks" or "compromised" pathways in N-glycosylation and subsequently allow for the development of strategies to improve glycosylation quality.

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Section: )supporting

confidence: 91%

Profiling ofN‐glycosylation gene expression in CHO cell fed‐batch cultures

Wong

Goh

et al. 2010

Biotech & Bioengineering

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“…Sialylation occurs at all stages of mammalian development and even embryonic stem cells already express sialylated glycoconjugates [57–59]. The expression of GNE mRNA, encoding the key enzyme of the sialic acid biosynthesis, strongly correlates with the occurrence of sialic acid.…”

Section: Regulation Of Gnementioning

confidence: 99%

“…For example, polysialylation of the neural cell adhesion molecule (NCAM) was affected in Gne deficient ES-cells, which could be restored with supplementation of ManNAc to the growth medium [28]. And recent studies of Gne deficient ES-cell showed that Gne , apart from its role in sialic acid synthesis, may also play a role in cell proliferation, gene expression, and cell differentiation [59]. …”

Section: Gne-opathiesmentioning

confidence: 99%

UDP-GlcNAc 2-Epimerase/ManNAc Kinase (GNE): A Master Regulator of Sialic Acid Synthesis

Hinderlich

Weidemann

Yardeni

et al. 2013

Topics in Current Chemistry

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“…Sialylation of glycoproteins and glycosphingolipids occurs late within the secretory apparatus by a family of Golgi-localized sialyltransferases that most commonly catalyze α2-3 or α2-6 linkages to galactose or the α2-8 linkage to sialic acid in oligo or polysialic acid structures [47]. This process is critical for mammalian development as sialylated glycans coat the surface of cells and perform a multitude of biological functions [47-49]. In addition, sialylated glycans on the surface play major roles in regulating immune and inflammatory responses and in host recognition by viruses, which often recognize specific sialic acid linkages to gain entry into cells [50].…”

Section: Introductionmentioning

confidence: 99%

Glycosylation and stem cells: Regulatory roles and application of iPSCs in the study of glycosylation‐related disorders

Berger¹,

Dookwah²,

Steet³

et al. 2016

BioEssays

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Glycosylation refers to the co- and post-translational modification of protein and lipids by monosaccharides or oligosaccharide chains. The surface of mammalian cells is decorated by a heterogeneous and highly complex array of protein and lipid linked glycan structures that vary significantly between different cell types, raising questions about their roles in development and disease pathogenesis. This review will begin by focusing on recent findings that define roles for cell surface protein and lipid glycosylation in pluripotent stem cells and their functional impact during normal development. Then, we will describe how patient derived induced pluripotent stem cells are being used to model human diseases such as congenital disorders of glycosylation. Collectively, these studies indicate that cell surface glycans perform critical roles in human development and disease.

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Lessons from GNE-deficient embryonic stem cells: sialic acid biosynthesis is involved in proliferation and gene expression

Cited by 35 publications

References 53 publications

Profiling ofN‐glycosylation gene expression in CHO cell fed‐batch cultures

Profiling ofN‐glycosylation gene expression in CHO cell fed‐batch cultures

UDP-GlcNAc 2-Epimerase/ManNAc Kinase (GNE): A Master Regulator of Sialic Acid Synthesis

Glycosylation and stem cells: Regulatory roles and application of iPSCs in the study of glycosylation‐related disorders

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