title: miR-2861 as novel HDAC5 inhibitor in CHO cells enhances productivity while maintaining product quality.
abstract: Histone deacetylase (HDAC) inhibitors have been exploited for years to improve recombinant protein expression in mammalian production cells. However, global HDAC inhibition is associated with negative effects on various cellular processes. microRNAs (miRNAs) have been shown to regulate gene expression in almost all eukaryotic cell types by controlling entire cellular pathways. Since miRNAs recently have gained much attention as next-generation cell engineering tool to improve Chinese hamster ovary (CHO) cell factories, we were interested if miRNAs are able to specifically repress HDAC expression in CHO cells to circumvent limitations of unspecific HDAC inhibition. We discovered a novel miRNA in CHO cells, miR-2861, which was shown to enhance productivity in various recombinant CHO cell lines. Furthermore, we demonstrate that miR-2861 might post-transcriptionally regulate HDAC5 in CHO cells. Intriguingly, siRNA-mediated HDAC5 suppression could be demonstrated to phenocopy pro-productive effects of miR-2861 in CHO cells. This supports the notion that miRNA-induced inhibition of HDAC5 may contribute to productivity enhancing effects of miR-2861. Furthermore, since product quality is fundamental to safety and functionality of biologics, we examined the effect of HDAC inhibition on critical product quality attributes. In contrast to unspecific HDAC inhibition using VPA, enforced expression of miR-2861 did not negatively influence antibody aggregation or N-glycosylation. Our findings highlight the superiority of miRNA-mediated inhibition of specific HDACs and present miR-2861 as novel cell engineering tool for improving CHO manufacturing cells.
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany. simon.fischer@hochschule-bc.de.|Faculty of Medicine, University of Ulm, Albert-Einstein-Allee 11, 89081 Ulm, Germany. simon.fischer@hochschule-bc.de.
authors.forename: Simon
authors.initials: S
authors.lastname: Fischer
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.
authors.forename: Albert Jesuran
authors.initials: AJ
authors.lastname: Paul
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.|Faculty of Natural Sciences, University of Ulm, Ulm, Germany.
authors.forename: Andreas
authors.initials: A
authors.lastname: Wagner
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.|Faculty of Natural Sciences, University of Ulm, Ulm, Germany.
authors.forename: Sven
authors.initials: S
authors.lastname: Mathias
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.|Faculty of Natural Sciences, University of Ulm, Ulm, Germany.
authors.forename: Melanie
authors.initials: M
authors.lastname: Geiss
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.|Faculty of Natural Sciences, University of Ulm, Ulm, Germany.
authors.forename: Franziska
authors.initials: F
authors.lastname: Schandock
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.|Faculty of Natural Sciences, University of Ulm, Ulm, Germany.
authors.forename: Martin
authors.initials: M
authors.lastname: Domnowski
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.|Faculty of Natural Sciences, University of Ulm, Ulm, Germany.
authors.forename: Jörg
authors.initials: J
authors.lastname: Zimmermann
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.
authors.forename: René
authors.initials: R
authors.lastname: Handrick
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.
authors.forename: Friedemann
authors.initials: F
authors.lastname: Hesse
authors.affiliation: Institute of Applied Biotechnology, University of Applied Sciences Biberach, Biberach, Germany.
authors.forename: Kerstin
authors.initials: K
authors.lastname: Otte
mesh_terms: D000818:Animals; D000906:Antibodies / Q000235:genetics / Q000378:metabolism; D016466:CHO Cells; D003412:Cricetulus; D005786:Gene Expression Regulation; D056572:Histone Deacetylase Inhibitors / Q000378:metabolism*; D060847:Metabolic Engineering; D035683:MicroRNAs / Q000378:metabolism*; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D000906:Antibodies; D056572:Histone Deacetylase Inhibitors; D035683:MicroRNAs; D011994:Recombinant Proteins
keywords: CHO; VPA; cell engineering; microRNA; monoclonal antibody; product quality