In Inflamed Intestinal Tissues and Epithelial Cells, Interleukin 22 Signaling Increases Expression of H19 Long Noncoding RNA, Which Promotes Mucosal Regeneration.
In Inflamed Intestinal Tissues and Epithelial Cells, Interleukin 22 Signaling Increases Expression of H19 Long Noncoding RNA, Which Promotes Mucosal Regeneration.
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在发炎的肠道组织和上皮细胞中，白介素22信号传导增加了H19长的非编码RNA的表达，从而促进粘膜再生。
DOI：
10.1053/j.gastro.2018.03.058
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发表时间：
2018-07
期刊：
影响因子：
29.4
通讯作者：
Tan XD
中科院分区：
文献类型：
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作者：
Geng H;Bu HF;Liu F;Wu L;Pfeifer K;Chou PM;Wang X;Sun J;Lu L;Pandey A;Bartolomei MS;De Plaen IG;Wang P;Yu J;Qian J;Tan XD
炎症影响肠上皮细胞的再生;长链非编码RNA（lncRNA）调节细胞功能，如增殖、分化和迁移。我们研究了lncRNA H19的机制，印迹母代表达的转录本（H19）调节再生的肠上皮细胞培养和小鼠模型的炎症。我们对脂多糖（LPS）诱导的脓毒症小鼠的肠组织进行了RNA-seq转录组分析，以确定与炎症相关的lncRNA;通过定量实时聚合酶链反应和原位杂交分析证实了与健康个体（对照）相比，脓毒症或葡聚糖硫酸钠（DSS）诱导的粘膜伤口愈合小鼠和溃疡性结肠炎患者的肠组织的结果。我们筛选了细胞因子诱导HT-29细胞和肠上皮细胞（IEC）中H19表达的能力，并证实了来自小鼠小肠的隐窝上皮类器官中的发现。将IEC与不同的信号转导抑制剂孵育，并测量对H19 lncRNA水平的影响。我们评估了给予LPS或DSS的H19Δ Ex 1/+小鼠与野生型同窝小鼠（对照小鼠）的肠上皮增殖或再生。使用慢病毒载体在IEC中过表达H19，并测量细胞增殖。我们进行了RNA反义纯化，RNA免疫沉淀，和荧光素酶报告基因分析，以研究H19在IEC的功能。在小鼠肠组织中lncRNA表达的RNA测序转录组分析中，我们发现H19 lncRNA的水平随着LPS暴露而显著变化。与对照组相比，溃疡性结肠炎患者、LPS诱导的多微生物败血症小鼠或DSS诱导的结肠炎小鼠的肠组织中H19 lncRNA水平增加。增加的H19 lncRNA定位于肠中的上皮细胞，无论Lgr 5信使RNA表达如何。IEC暴露于白细胞介素22（IL 22）随时间和剂量增加H19 lncRNA水平，这需要STAT 3和蛋白激酶A活性。IL 22在6小时内诱导小鼠肠上皮类器官中H19的表达。暴露于IL 22增加了来自对照小鼠的肠上皮类器官的生长，但不增加H19Δ Ex 1/+小鼠的生长。H19在HT-29细胞中的过表达促进其增殖。与对照小鼠相比，H19Δ Ex 1/+小鼠停用DSS后肠粘膜愈合更慢。LPS刺激后，H19Δ Ex 1/+小鼠的腺上皮细胞比对照小鼠的腺上皮细胞增殖缓慢。H19 lncRNA与p53和抑制细胞增殖的microRNA结合，包括microRNA 34 a和let-7; H19 lncRNA结合阻断了它们的功能，导致促进上皮再生的基因表达增加。lncRNA H19的水平在来自小鼠和患者的发炎肠组织中增加。炎性细胞因子IL 22诱导IEC中H19的表达，这是肠上皮细胞增殖和粘膜愈合所需的。H19 lncRNA似乎抑制p53蛋白和microRNA 34 a和let-7以促进IEC增殖和上皮再生。
Inflammation affects regeneration of the intestinal epithelia; long noncoding RNAs (lncRNAs) regulate cell functions such as proliferation, differentiation, and migration. We investigated the mechanisms by which the lncRNA H19, imprinted maternally expressed transcript (H19) regulates regeneration of intestinal epithelium using cell cultures and mouse models of inflammation. We performed RNA-seq transcriptome analyses of intestinal tissues from mice with lipopolysaccharide (LPS)-induced sepsis to identify lncRNAs associated with inflammation; findings were confirmed by quantitative real-time polymerase chain reaction and in situ hybridization analyses of intestinal tissues from mice with sepsis or dextran sulfate sodium (DSS)-induced mucosal wound healing and patients with ulcerative colitis compared to healthy individuals (controls). We screened cytokines for their ability to induce expression of H19 in HT-29 cells and intestinal epithelial cells (IECs), and confirmed findings in crypt epithelial organoids derived from mouse small intestine. IECs were incubated with different signal transduction inhibitors and effects on H19 lncRNA levels were measured. We assessed intestinal epithelial proliferation or regeneration in H19ΔEx1/+ mice given LPS or DSS vs wild-type littermates (control mice). H19 was overexpressed in IECs using lentiviral vectors and cell proliferation was measured. We performed RNA antisense purification, RNA immunoprecipitation, and luciferase reporter assays to study functions of H19 in IECs. In RNA-sequencing transcriptome analysis of lncRNA expression in intestinal tissues from mice, we found that levels of H19 lncRNA changed significantly with LPS exposure. Levels of H19 lncRNA increased in intestinal tissues of patients with ulcerative colitis, mice with LPS-induced and polymicrobial sepsis, or mice with DSS-induced colitis, compared with controls. Increased H19 lncRNA localized to epithelial cells in the intestine, regardless of Lgr5 messenger RNA expression. Exposure of IECs to interleukin 22 (IL22) increased levels of H19 lncRNA with time and dose, which required STAT3 and protein kinase A activity. IL22 induced expression of H19 in mouse intestinal epithelial organoids within 6 hours. Exposure to IL22 increased growth of intestinal epithelial organoids derived from control mice, but not H19ΔEx1/+ mice. Overexpression of H19 in HT-29 cells increased their proliferation. Intestinal mucosa healed more slowly after withdrawal of DSS from H19ΔEx1/+ mice vs control mice. Crypt epithelial cells from H19ΔEx1/+ mice proliferated more slowly than those from control mice after exposure to LPS. H19 lncRNA bound to p53 and microRNAs that inhibit cell proliferation, including microRNA 34a and let-7; H19 lncRNA binding blocked their function, leading to increased expression of genes that promote regeneration of the epithelium. The level of lncRNA H19 is increased in inflamed intestinal tissues from mice and patients. The inflammatory cytokine IL22 induces expression of H19 in IECs, which is required for intestinal epithelial proliferation and mucosal healing. H19 lncRNA appears to inhibit p53 protein and microRNA 34a and let-7 to promote proliferation of IECs and epithelial regeneration.