title: Targeted epigenetic editing of SPDEF reduces mucus production in lung epithelial cells.
abstract: Airway mucus hypersecretion contributes to the morbidity and mortality in patients with chronic inflammatory lung diseases. Reducing mucus production is crucial for improving patients' quality of life. The transcription factor SAM-pointed domain-containing Ets-like factor (SPDEF) plays a critical role in the regulation of mucus production and, therefore, represents a potential therapeutic target. This study aims to reduce lung epithelial mucus production by targeted silencingSPDEFusing the novel strategy, epigenetic editing. Zinc fingers and CRISPR/dCas platforms were engineered to target repressors (KRAB, DNA methyltransferases, histone methyltransferases) to theSPDEFpromoter. All constructs were able to effectively suppress bothSPDEFmRNA and protein expression, which was accompanied by inhibition of downstream mucus-related genes [anterior gradient 2 (AGR2), mucin 5AC (MUC5AC)]. For the histone methyltransferase G9A, and not its mutant or other effectors, the obtained silencing was mitotically stable. These results indicate efficientSPDEFsilencing and downregulation of mucus-related gene expression by epigenetic editing, in human lung epithelial cells. This opens avenues for epigenetic editing as a novel therapeutic strategy to induce long-lasting mucus inhibition.
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, The Netherlands.|University of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, The Netherlands.|Tianjin Medical University, School of Basic Medical Sciences, Department of Biochemistry and Molecular Biology, Department of Immunology, Tianjin, China; and.
authors.forename: Juan
authors.identifier: 0000-0001-7350-9578
authors.initials: J
authors.lastname: Song
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, The Netherlands.
authors.forename: David
authors.initials: D
authors.lastname: Cano-Rodriquez
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, The Netherlands.
authors.forename: Melanie
authors.initials: M
authors.lastname: Winkle
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, The Netherlands.
authors.forename: Rutger A F
authors.initials: RA
authors.lastname: Gjaltema
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, The Netherlands.
authors.forename: Désirée
authors.initials: D
authors.lastname: Goubert
authors.affiliation: Institute of Biochemistry, Faculty of Chemistry, University of Stuttgart, Stuttgart, Germany.
authors.forename: Tomasz P
authors.initials: TP
authors.lastname: Jurkowski
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, The Netherlands.|University of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, The Netherlands.
authors.forename: Irene H
authors.initials: IH
authors.lastname: Heijink
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, The Netherlands.
authors.forename: Marianne G
authors.initials: MG
authors.lastname: Rots
authors.affiliation: University of Groningen, University Medical Center Groningen, Department of Pathology and Medical Biology, Groningen, The Netherlands; m.n.hylkema@umcg.nl.|University of Groningen, University Medical Center Groningen, Groningen Research Institute for Asthma and COPD, Groningen, The Netherlands.
authors.forename: Machteld N
authors.identifier: 0000-0002-6732-8903
authors.initials: MN
authors.lastname: Hylkema
mesh_terms: D001483:Base Sequence; D002460:Cell Line; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics / Q000378:metabolism; D019175:DNA Methylation / Q000235:genetics; D000090205:DNA Methyltransferase 3A; D015536:Down-Regulation / Q000235:genetics; D044127:Epigenesis, Genetic; D004847:Epithelial Cells / Q000378:metabolism*; D000072669:Gene Editing; D020868:Gene Silencing; D006649:Histocompatibility Antigens / Q000235:genetics / Q000378:metabolism; D011495:Histone-Lysine N-Methyltransferase / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008168:Lung / Q000166:cytology*; D008954:Models, Biological; D055271:Mucin 5AC / Q000378:metabolism; D009093:Mucus / Q000378:metabolism*; D011401:Promoter Regions, Genetic / Q000235:genetics; D000072417:Protein Domains; D050783:Proto-Oncogene Proteins c-ets / Q000235:genetics* / Q000378:metabolism; D016335:Zinc Fingers
chemical_list: C000717967:DNMT3A protein, human; D006649:Histocompatibility Antigens; D055271:Mucin 5AC; D050783:Proto-Oncogene Proteins c-ets; C403974:SPDEF protein, human; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; C445405:EHMT2 protein, human; D011495:Histone-Lysine N-Methyltransferase
keywords: DNA methylation; SPDEF; epigenetic editing; mucus production