Promoter hypomethylation of Toll-like receptor-2 gene is associated with increased proinflammatory response toward bacterial peptidoglycan in cystic fibrosis bronchial epithelial cells.
Cystic fibrosis (CF) is the most common lethal inherited disorder caused by mutation in the gene encoding CF transmembrane regulator (CFTR). The clinical course of CF is characterized by recurrent pulmonary infections and chronic inflammation. Here, we show that toll-like receptor-2 (TLR2) expression and response were strongly enhanced in the human CF bronchial epithelial cell line, CFBE41o-. Treatment of the cells with 5-azacytidine decreased the promoter methylation within TLR2 proximal promoter and increased endogenous expression of TLR2 in non-CF 16HBE14o- cells, suggesting that TLR2 expression is epigenetically regulated by CpG methylation. Moreover, bisulfite sequence analysis revealed that TLR2 promoters were highly demethylated in CFBE41o- cells, implying that decreased methylation of the TLR2 promoter is responsible for CF-related up-regulation of TLR2. Finally, stable expression of WT-CFTR in CFBE41o- cells (CFBE41o-/WT-CFTR cells) reduced TLR2 expression and the response to its ligand peptidoglycan (PGN), implying a causal relationship between CFTR dysfunction and TLR2 up-regulation. Consistent with reduced expression of TLR2 in CFBE41o-/WT-CFTR cells, CpG methylation was increased in CFBE41o-/WT-CFTR cells. Taken together, our results demonstrate that TLR2 expression is epigenetically up-regulated in CF bronchial epithelial cells and suggest that TLR2 overexpression or prolonged activation of TLR2 signaling might be critical in CF pathogenesis.
Shuto
Tsuyoshi
T
Department of Molecular Medicine, Graduate School of Pharmaceutical Science, Kumamoto University, Kumamoto, Japan.
Furuta
Takashi
T
Oba
Mariko
M
Xu
Haidong
H
Li
Jian-Dong
JD
Cheung
Judy
J
Gruenert
Dieter C
DC
Uehara
Akiko
A
Suico
Mary Ann
MA
Okiyoneda
Tsukasa
T
Kai
Hirofumi
H
D001374:Azacitidine; D002478:Cells, Cultured; D003550:Cystic Fibrosis / Q000378:metabolism* / Q000382:microbiology / Q000473:pathology; D019005:Cystic Fibrosis Transmembrane Conductance Regulator / Q000235:genetics; D019175:DNA Methylation; D004721:Endoplasmic Reticulum / Q000378:metabolism; D004847:Epithelial Cells / Q000378:metabolism*; D005786:Gene Expression Regulation; D006801:Humans; D007249:Inflammation / Q000276:immunology / Q000473:pathology; D010457:Peptidoglycan / Q000276:immunology*; D011401:Promoter Regions, Genetic / Q000235:genetics*; D020545:Respiratory Mucosa / Q000166:cytology*; D013211:Staphylococcus aureus / Q000378:metabolism; D051195:Toll-Like Receptor 2 / Q000235:genetics*; D015854:Up-Regulation
D010457:Peptidoglycan; C495342:TLR2 protein, human; D051195:Toll-Like Receptor 2; D019005:Cystic Fibrosis Transmembrane Conductance Regulator; D001374:Azacitidine