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Genome-wide DNA methylation and transcriptome expression profiles of peripheral blood mononuclear cells in patients with systemic sclerosis with interstitial lung disease.

OBJECTIVES: This study aims to investigate the genome-wide DNA methylation and transcriptome expression profiles of peripheral blood mononuclear cells (PBMCs) in patients with systemic sclerosis (SSc) with interstitial lung disease (ILD), and to analyze the effects of DNA methylation on Wnt/β-catenin and chemokine signaling pathways.

METHODS: PBMCs were collected from 19 patients with SSc (SSc group) and 18 healthy persons (control group). Among SSc patients, there were 10 patients with ILD (SSc with ILD subgroup) and 9 patients without ILD (SSc without ILD subgroup). The genome-wide DNA methylation and gene expression level were analyzed by using Illumina 450K methylation chip and Illumina HT-12 v4.0 gene expression profiling chip. The effect of DNA methylation on Wnt/β-catenin and chemokine signal pathways was investigated.

RESULTS: Genome-wide DNA methylation analysis identified 71 hypermethylated CpG sites and 98 hypomethylated CpG sites in the SSc with ILD subgroup compared with the SSc without ILD subgroup. Transcriptome analysis distinguished 164 upregulated genes and 191 downregulated genes in the SSc with ILD subgroup as compared with the SSc without ILD subgroup. In PBMCs of the SSc group, 35 genes in Wnt/β-catenin signaling pathway were hypomethylated, while frizzled-1 ( FZD1 ), mitogen-activated protein kinase 9 ( MAPK9 ), mothers against DPP homolog 2 ( SMAD2 ), transcription factor 7-like 2 ( TCF7L2 ), and wingless-type MMTV integration site family, member 5B ( WNT5B ) mRNA expressions were upregulated as compared with the control group (all P <0.05). Compared with the SSc without ILD subgroup, the mRNA expressions of dickkopf homolog 2 ( DKK2 ), FZD1 , MAPK9 were upregulated in the SSc with ILD subgroup, but the differences were not statistically significant (all P >0.05). In PBMCs of the SSc group, 38 genes in chemokine signaling pathway were hypomethylated, while β-arrestin 1 ( ARRB1 ), C-X-C motif chemokine ligand 10 ( CXCL10 ), C-X-C motif chemokine ligand 16 ( CXCL16 ), FGR , and neutrophil cytosolic factor 1C ( NCF1C ) mRNA expressions were upregulated as compared with the control group (all P <0.05). Compared with the SSc without ILD subgroup, the mRNA expressions of ARRB1 , CXCL10 , CXCL16 were upregulated in the SSc with ILD subgroup, but the differences were not statistically significant (all P >0.05).

CONCLUSIONS: There are differences in DNA methylation and transcriptome profiles between SSc with ILD and SSc without ILD. The expression levels of multiple genes in Wnt/β- catenin and chemokine signaling pathways are upregulated, which might be associatea with the pathogenesis of SSc.

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