Journal
BIOTECHNOLOGY AND BIOENGINEERING
Volume 119, Issue 3, Pages 832-844Publisher
WILEY
DOI: 10.1002/bit.28015
Keywords
Chinese hamster ovary (CHO) cells; microRNA; monoclonal antibody; N-glycosylation; sialylation
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A study identified a recombinant CHO cell line with unusually high levels of NGNA sialylation, and the root cause was found through comprehensive multi-OMICs experimental analyses, involving the spontaneous expression of the CMAH gene, microRNA miR-111 regulation, and a point mutation in the SDK1 gene.
Chinese hamster ovary (CHO) cells are known not to express appreciable levels of the sialic acid residue N-glycolylneuraminic acid (NGNA) on monoclonal antibodies. However, we actually have identified a recombinant CHO cell line expressing an IgG with unusually high levels of NGNA sialylation (>30%). Comprehensive multi-OMICs based experimental analyses unraveled the root cause of this atypical sialylation: (1) expression of the cytidine monophosphate-N-acetylneuraminic acid hydroxylase (CMAH) gene was spontaneously switched on, (2) CMAH mRNA showed an anti-correlated expression to the newly discovered Cricetulus griseus (cgr) specific microRNA cgr-miR-111 and exhibits two putative miR-111 binding sites, (3) miR-111 expression depends on the transcription of its host gene SDK1, and (4) a single point mutation within the promoter region of the sidekick cell adhesion molecule 1 (SDK1) gene generated a binding site for the transcriptional repressor histone H4 transcription factor HINF-P. The resulting transcriptional repression of SDK1 led to a downregulation of its co-expressed miR-111 and hence to a spontaneous upregulation of CMAH expression finally increasing NGNA protein sialylation.
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