Wnt signaling regulates immunomodulatory functions during infection and inflammation. Employing NCCIT and HCT116 cells, having high endogenous Wnt signaling, we observed elevated levels of low-density lipoprotein receptor-related protein 5/6 (LRP5/6) and Frizzled class receptor 10 (FZD10) and increases in β-catenin, doublecortin-like kinase 1 (DCLK1), CD44 molecule (CD44), and aldehyde dehydrogenase 1 family member A1 (ALDH1A1). siRNA-induced knockdown of these receptors antagonized TOPflash reporter activity and spheroid growth in vitro and elevated Wnt-inhibitory factor 1 (WIF1) activity. Elevated mRNA and protein levels of LRP5/6 and FZD10 paralleled expression of WNT2b and WNT4 in colonic crypts at days 6 and 12 post-infection with Citrobacter rodentium (CR) and tended to decline at days 20-34. The CR mutant escV or the tankyrase inhibitor XAV939 attenuated these responses. A three-dimensional organoid assay in colonic crypts isolated from CR-infected mice revealed elevated levels of LRP5/6 and FZD10 and β-catenin co-localization with enhancer of zeste 2 polycomb repressive complex 2 subunit (EZH2). Co-immunoprecipitation in the membrane fraction revealed that axin associates with LRP5/6 in CR-infected crypts, and this association was correlated with increased β-catenin. Colon tumors from either CR-infected ApcP Min/+ or azoxymethane/dextran sodium sulfate (AOM/DSS)-treated mice had high LRP5/6 or FZD10 levels, and chronic Notch blockade through the γ-secretase inhibitor dibenzazepine down-regulated LRP5/6 and FZD10 expression. In CR-responsive CT-26 cells, siRNA-induced LRP5/6 or FZD10 knockdown antagonized TOPflash reporter activity. Elevated miR-153-3p levels correlated with LRP5/6 and FZD10, and miR-153-3p sequestration via a plasmid-based miR inhibitor system attenuated Wnt signaling. We conclude that infection-induced signals from the plasma membrane epigenetically regulate Wnt signaling.

中文翻译：

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在质膜上产生的感染诱导信号在体外和体内表观遗传调控Wnt信号传导。

Wnt信号传导在感染和炎症过程中调节免疫调节功能。使用具有高内源性Wnt信号传导的NCCIT和HCT116细胞，我们观察到低密度脂蛋白受体相关蛋白5/6（LRP5 / 6）和卷曲蛋白类受体10（FZD10）的水平升高，β-catenin，doublecortin-像激酶1（DCLK1），CD44分子（CD44）和醛脱氢酶1家族成员A1（ALDH1A1）。siRNA诱导的这些受体的敲低在体外拮抗TOPflash报告基因的活性和球状体的生长，并提高Wnt抑制因子1（WIF1）的活性。LRP5 / 6和FZD10的mRNA和蛋白水平升高，在感染鼠尾草柠檬酸杆菌（CR）后第6和12天与结肠隐窝中WNT2b和WNT4的表达平行，并在20-34天趋于下降。CR突变escV或tankyrase抑制剂XAV939减弱了这些反应。在从CR感染的小鼠中分离出的结肠隐窝中进行的三维类器官测定表明LRP5 / 6和FZD10和β-catenin与zeste 2聚梳抑制复合物2亚基（EZH2）的增强子共定位。膜级分中的共免疫沉淀表明，在CR感染的隐窝中，毒素与LRP5 / 6相关，并且这种相关与β-catenin的增加有关。来自CR感染的ApcP Min / +或乙氧基甲烷/葡聚糖硫酸钠（AOM / DSS）处理的小鼠的结肠肿瘤具有较高的LRP5 / 6或FZD10水平，并且通过γ-分泌酶抑制剂地苯扎西平下调的LRP5 / 6和FZD10表达。在CR响应的CT-26细胞中，siRNA诱导的LRP5 / 6或FZD10敲低可拮抗TOPflash报告基因的活性。升高的miR-153-3p水平与LRP5 / 6和FZD10相关，并且通过基于质粒的miR抑制剂系统隔离miR-153-3p会减弱Wnt信号传导。我们得出的结论是，感染引起的质膜表观遗传信号调控Wnt信号传导。