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

Juan Song, David Cano-Rodriquez, Melanie Winkle, Rutger A. F. Gjaltema, Desiree Goubert, Tomasz P. Jurkowski, Irene H. Heijink, Marianne G. Rots, Machteld N. Hylkema*

*Corresponding author for this work

Research output: Contribution to journalArticleAcademicpeer-review

24 Citations (Scopus)
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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 silencing SPDEF using the novel strategy, epigenetic editing. Zinc fingers and CRISPR/dCas platforms were engineered to target repressors (KRAB, DNA methyltransferases, histone methyltransferases) to the SPDEF promoter. All constructs were able to effectively suppress both SPDEF mRNA 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 efficient SPDEF 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.

Original languageEnglish
Pages (from-to)L334-L347
Number of pages14
JournalAmerican Journal of Physiology - Lung Cellular and Molecular Physiology
Volume312
Issue number3
Early online date20-Dec-2016
DOIs
Publication statusPublished - 1-Mar-2017

Keywords

  • SPDEF
  • epigenetic editing
  • mucus production
  • DNA methylation
  • DNA METHYLATION
  • AIRWAY EPITHELIUM
  • SPUTUM PRODUCTION
  • BREAST-CANCER
  • TRANSCRIPTION
  • EXPRESSION
  • GENES
  • COPD
  • DIFFERENTIATION
  • DYNAMICS

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