Targeted epigenetic editing of SPDEF reduces mucus production in lung epithelial cells

• Published in: American journal of physiology. Lung cellular and molecular physiology Vol. 312; no. 3; pp. L334 - L347
• Main Authors: Song, Juan, Cano-Rodriquez, David, Winkle, Melanie, Gjaltema, Rutger A F, Goubert, Désirée, Jurkowski, Tomasz P, Heijink, Irene H, Rots, Marianne G, Hylkema, Machteld N
• Format: Journal Article
• Language: English
• Published: United States American Physiological Society 01.03.2017
• Subjects: Base Sequence; Body fluids; Cell Line; DNA (Cytosine-5-)-Methyltransferases - genetics; DNA (Cytosine-5-)-Methyltransferases - metabolism; DNA Methylation - genetics; Down-Regulation - genetics; Epigenesis, Genetic; Epigenetics; Epithelial Cells - metabolism; Gene Editing; Gene expression; Gene Silencing; Histocompatibility Antigens - genetics; Histocompatibility Antigens - metabolism; Histone-Lysine N-Methyltransferase - genetics; Histone-Lysine N-Methyltransferase - metabolism; Humans; Lung - cytology; Lung diseases; Models, Biological; Mucin 5AC - metabolism; Mucus - metabolism; Physiology; Promoter Regions, Genetic - genetics; Protein Domains; Proto-Oncogene Proteins c-ets - genetics; Proto-Oncogene Proteins c-ets - metabolism; Quality of life; Zinc Fingers; epigenetic editing; DNA methylation; mucus production; SPDEF
• ISSN: 1040-0605; 1522-1504
• DOI: 10.1152/ajplung.00059.2016

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 ( ) 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 silencing using the novel strategy, epigenetic editing. Zinc fingers and CRISPR/dCas platforms were engineered to target repressors (KRAB, DNA methyltransferases, histone methyltransferases) to the promoter. All constructs were able to effectively suppress both mRNA and protein expression, which was accompanied by inhibition of downstream mucus-related genes [anterior gradient 2 ( ), mucin 5AC ( )]. For the histone methyltransferase G9A, and not its mutant or other effectors, the obtained silencing was mitotically stable. These results indicate efficient silencing 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.