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
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.
Am J Physiol Lung Cell Mol Physiol. 2017 Mar 1;312(3):L334-L347. doi: 10.1152/ajplung.00059.2016. Epub 2016 Dec 23.
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.
气道黏液高分泌会导致慢性炎症性肺疾病患者的发病和死亡。减少黏液分泌对于提高患者生活质量至关重要。转录因子含SAM结构域的Ets样因子（）在黏液分泌调节中起关键作用，因此是一个潜在的治疗靶点。本研究旨在通过使用新型策略——表观遗传编辑，靶向沉默来减少肺上皮细胞黏液分泌。设计锌指核酸酶和CRISPR/dCas平台，将阻遏物（KRAB、DNA甲基转移酶、组蛋白甲基转移酶）靶向到启动子。所有构建体均能有效抑制mRNA和蛋白表达，同时抑制下游黏液相关基因[前梯度2（）、黏蛋白5AC（）]。对于组蛋白甲基转移酶G9A，而非其突变体或其他效应物，所获得的沉默在有丝分裂过程中是稳定的。这些结果表明，在人肺上皮细胞中，通过表观遗传编辑可有效沉默并下调黏液相关基因的表达。这为表观遗传编辑作为一种诱导持久黏液抑制的新型治疗策略开辟了道路。
