UHRF1 deficiency exacerbates intestinal inflammation by epigenetic modulation of NPY1R gene methylation
Yanan Han, Lina Sun, Yanxing Liu, Xiaohui Zhang, Hao Liu, Haohao Zhang, Xiaoxia Ren, Fenfan Wang, Huafeng Fan, Jie Chen, Dan Liu, Daiming Fan, Yuanyuan Lu, Xue Bai, Ying Fang, Kaichun Wu, Xiaodi Zhao
Yanan Han, Lina Sun, Yanxing Liu, Xiaohui Zhang, Hao Liu, Haohao Zhang, Xiaoxia Ren, Fenfan Wang, Huafeng Fan, Jie Chen, Dan Liu, Daiming Fan, Yuanyuan Lu, Xue Bai, Ying Fang, Kaichun Wu, Xiaodi Zhao
View: Text | PDF
Research Article Gastroenterology Inflammation

UHRF1 deficiency exacerbates intestinal inflammation by epigenetic modulation of NPY1R gene methylation

Abstract

Epigenetic modifications play a crucial role in the pathogenesis of inflammatory bowel disease (IBD) by mediating gene-environment interactions. We previously showed that UHRF1, a central regulator of DNA methylation, contributes to cancer progression; however, its function in IBD remains poorly understood. Here, we revealed that UHRF1 was frequently reduced in inflamed tissues of patients with IBD and that its deficiency exacerbated intestinal epithelial cell (IEC) damage. Through a multilevel approach incorporating human cell models and an intestinal epithelial-specific Uhrf1-KO mouse model, we established UHRF1 as a key mitigator of IBD progression. Mechanistically, UHRF1 bound to the NPY1R promoter, promoted its methylation, and led to transcriptional suppression. The NPY1R upregulation resulting from UHRF1 deficiency attenuated cAMP/PKA/CREB signaling in IECs, thereby enhancing NF-κB activation and subsequent proinflammatory responses, which compromised intestinal epithelial barrier integrity. Furthermore, we identified miR-141 as a negative regulator of NPY1R, highlighting its potential as a therapeutic agent. Collectively, our results identified the UHRF1/NPY1R regulatory axis as a critical epigenetic mechanism in intestinal inflammation and underscored its dual promise for IBD diagnostics and therapy.

Authors

Yanan Han, Lina Sun, Yanxing Liu, Xiaohui Zhang, Hao Liu, Haohao Zhang, Xiaoxia Ren, Fenfan Wang, Huafeng Fan, Jie Chen, Dan Liu, Daiming Fan, Yuanyuan Lu, Xue Bai, Ying Fang, Kaichun Wu, Xiaodi Zhao

×

Figure 3

UHRF1 represses NPY1R expression through promoter methylation in IECs.

Options: View larger image (or click on image) Download as PowerPoint
UHRF1 represses NPY1R expression through promoter methylation in IECs. (...
(A) Venn diagram for upregulated and hypomethylated genes in NCM460 cells infected with shUHRF1 or shCtrl, and Gene Ontology (GO) enrichment analysis of genes common to these 2 gene sets. (B) Representative staining images and correlation analysis of UHRF1 and NPY1R expression in colonic tissues from patients with UC and healthy donors (n = 25). Scale bar: 50 μm. (C) mRNA expression and correlation analysis of UHRF1 and NPY1R in paired inflamed and noninflamed colon tissue samples from patients with UC (n = 15). (D) Representative staining images and correlation analysis of UHRF1 and NPY1R in Uhrf1IEC(+/+) (n = 16) and Uhrf1IEC(+/−) (n = 15) mice. Scale bar: 50 μm. (E) Representative staining images and quantification of NPY1R in colon tissues from mice treated with DSS and sacrificed at different time points (n = 5). Scale bar: 50 μm. (F) Representative staining images and quantification of NPY1R in colon tissues from mice treated with AIEC or controls (n = 7). Scale bar: 50 μm. (G) mRNA and protein expression of NPY1R in NCM460 and FHC cells transfected with siUHRF1 or siCtrl. (H) mRNA and protein expression of NPY1R in NCM460 and FHC cells transfected with siDNMT1 or siCtrl. (I) Upper: Schematic depicts the CpG island (purple bar) and CpG sites (purple ticks) in the NPY1R promoter. Lower: Methylation status of specific CpG sites was assessed by bisulfite cloning and sequencing. (J) Analysis of ChIP-enriched DNA from the NPY1R promoter CpG island by conventional PCR (left) and qPCR (right) in NCM460 cells infected with shUHRF1 or shCtrl. P values were determined by χ2 test (B, C [right], and D), 1-way ANOVA (E, G, H, and J), or 2-tailed Student’s t tests (C [left and middle] and F). Data are shown as mean ± SD. *P < 0.05, **P < 0.01.