Targeted epigenetic silencing of UCHL1 expression suppresses collagen-1 production in human lung epithelial cells

Abstract

Ubiquitin carboxyl-terminal hydrolase L1 (UCHL1) is highly expressed in smokers, but little is known about the molecular mechanism of UCHL1 in airway epithelium and its possible role in affecting extracellular matrix (ECM) remodelling in the underlying submucosa. Since cigarette smoking is a major cause of lung diseases, we studied its effect on UCHL1 expression and DNA methylation patterns in human bronchial epithelial cells, obtained after laser capture micro-dissection (LCM) or isolated from residual tracheal/main stem bronchial tissue. Targeted regulation of UCHL1 expression via CRISPR/dCas9 based-epigenetic editing was used to explore the function of UCHL1 in lung epithelium. Our results show that cigarette smoke extract (CSE) stimulated the expression of UCHL1 in vitro . The methylation status of the UCHL1 gene was negatively associated with UCHL1 transcription in LCM-obtained airway epithelium at specific sites. Treatment with a UCHL1 inhibitor showed that the TGF-β1-induced upregulation of the ECM gene COL1A1 can be prevented by the inhibition of UCHL1 activity in cell lines. Furthermore, upon downregulation of UCHL1 by epigenetic editing using CRISPR/dCas-EZH2, mRNA expression of COL1A1 and fibronectin was reduced. In conclusion, we confirmed higher UCHL1 expression in current smokers compared to non- and ex-smokers, and induced downregulation of UCHL1 by epigenetic editing. The subsequent repression of genes encoding ECM proteins suggest a role for UCHL1 as a therapeutic target in fibrosis-related disease.

Nicotinamide riboside Induced Energy Stress and Metabolic Reprogramming in BEAS-2B Cells

“…In parallel, NR exposure led to decreased abundance of proteins involved in the folding, aggregation, or turnover of proteins, in l -serine biosynthesis, mRNA transcription, posttranscriptional gene silencing, double-strand break repair, and regulation of protein phosphorylation (Figures c, , S5 and Table S6). Among the 12 downregulated proteins described here, 11 have been shown to be upregulated in different types of cancer. − Downregulation of these proteins may have a role in slowing the growth and inducing apoptosis of the NR exposed cells, despite the shift to glycolytic metabolism. To the best of our knowledge, this is the first report showing the modulation of the abundance of the described sets of proteins by NR exposure.…”

“…Except for cellular tumor antigen p53 (TP53), the other 11 proteins have been shown to be upregulated in different types of cancer (Table S6). − Six of these proteins are involved in the folding, aggregation, or turnover of proteins [hypoxia up-regulated protein 1 (HYOU1), heat shock 70 kDa protein 1A or 1B (HSPA1A or 1B), protein disulfide-isomerase (P4HB), prolyl 4-hydroxylase subunit alpha-2 (P4HA2), serpin H1 (SERPINH1), ubiquitin carboxyl-terminal hydrolase isozyme L1 (UCHL1)]. Two are involved in l -serine biosynthesis [phosphoserine aminotransferase (PSAT1), phosphoserine phosphatase (PSPH)].…”

Evodiamine inhibits EPRS expression to regulate glutamate metabolism and proliferation of oral squamous cell carcinoma cells

“…Urgent exploration of novel therapeutic strategies is imperative. Recent years have witnessed growing interest in natural compounds derived from traditional Chinese medicine due to their potential anticancer properties 3 . Among these, evodiamine has emerged as a widely studied natural compound, showcasing significant efficacy in inhibiting tumor growth 4 …”

“…Recent years have witnessed growing interest in natural compounds derived from traditional Chinese medicine due to their potential anticancer properties. 3 Among these, evodiamine has emerged as a widely studied natural compound, showcasing significant efficacy in inhibiting tumor growth. 4 Evodiamine, a natural indole alkaloid extracted from the Evodia ruticarpa plant, has demonstrated potent anticancer activity against a range of tumors.…”