MicroRNA-449a overexpression, reduced NOTCH1 signals and scarce goblet cells characterize the small intestine of celiac patients.
MicroRNA-449a overexpression, reduced NOTCH1 signals and scarce goblet cells characterize the small intestine of celiac patients.
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DOI：
10.1371/journal.pone.0029094
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发表时间：
2011
期刊：
影响因子：
3.7
通讯作者：
Sacchetti L
中科院分区：
文献类型：
--
作者：
Capuano M;Iaffaldano L;Tinto N;Montanaro D;Capobianco V;Izzo V;Tucci F;Troncone G;Greco L;Sacchetti L
miRNAs在包括自身免疫性疾病在内的多种生理和病理条件下调节基因表达中发挥相关作用。miRNAs在小鼠肠上皮细胞的分化和功能中也很重要。我们的研究旨在寻找基于miRNA的腹腔小肠基因表达的调节，特别是参与细胞肠分化/增殖机制的基因。在儿科（那不勒斯费德里科二世大学）招募了40名儿童（20名活动性CD，9名无麸质饮食（GFD），11名对照）。通过TaqMan低密度阵列定量365种人miRNA的表达。我们使用生物信息学来预测miRNAs的推定靶基因并选择生物学途径。通过免疫组织化学检测CD和对照儿童小肠中NOTCH 1、HES 1、KLF 4、MUC-2、Ki 67和β-catenin蛋白的存在。在CD和对照儿童之间，大约20%的检测miRNAs的表达不同。我们发现高水平的miR-449 a靶向并减少HEK-293细胞中的NOTCH 1和KLF 4。与对照组相比，活动性CD儿童和GFD儿童小肠中的NOTCH 1、KLF 4信号和杯状细胞数量较低，而CD患者隐窝中的细胞核β-连环蛋白染色（作为WNT通路激活的标志）和Ki 67染色（作为增殖的标志）均高于对照组。总之，我们首次证明了CD患者小肠中的miRNA介导的基因调控。我们还强调了我们的患者中NOTCH 1途径的减少，无论疾病是否活跃。我们认为，NOTCH途径可以组成性改变，在腹腔小肠，并可以驱动增加增殖和减少分化的肠细胞向分泌杯状细胞谱系。
MiRNAs play a relevant role in regulating gene expression in a variety of physiological and pathological conditions including autoimmune disorders. MiRNAs are also important in the differentiation and function of the mouse intestinal epithelium. Our study was aimed to look for miRNA-based modulation of gene expression in celiac small intestine, and particularly for genes involved in cell intestinal differentiation/proliferation mechanisms. A cohort of 40 children (20 with active CD, 9 on a gluten-free diet (GFD), and 11 controls), were recruited at the Paediatrics Department (University of Naples Federico II). The expression of 365 human miRNAs was quantified by TaqMan low-density arrays. We used bioinformatics to predict putative target genes of miRNAs and to select biological pathways. The presence of NOTCH1, HES1, KLF4, MUC-2, Ki67 and beta-catenin proteins in the small intestine of CD and control children was tested by immunohistochemistry. The expression of about 20% of the miRNAs tested differed between CD and control children. We found that high miR-449a levels targeted and reduced both NOTCH1 and KLF4 in HEK-293 cells. NOTCH1, KLF4 signals and the number of goblet cells were lower in small intestine of children with active CD and in those on a GFD than in controls, whereas more nuclear beta-catenin staining, as a sign of the WNT pathway activation, and more Ki67 staining, as sign of proliferation, were present in crypts from CD patients than in controls. In conclusion we first demonstrate a miRNA mediated gene regulation in small intestine of CD patients. We also highlighted a reduced NOTCH1 pathway in our patients, irrespective of whether the disease was active or not. We suggest that NOTCH pathway could be constitutively altered in the celiac small intestine and could drive the increased proliferation and the decreased differentiation of intestinal cells towards the secretory goblet cell lineage.
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影响因子：
29.4
作者：
McKenna LB;Schug J;Vourekas A;McKenna JB;Bramswig NC;Friedman JR;Kaestner KH
通讯作者：
Kaestner KH
影响因子：
2.7
作者：
Ghaleb AM;McConnell BB;Kaestner KH;Yang VW
通讯作者：
Yang VW
影响因子：
3.5
作者：
Ciccocioppo, R;Finamore, A;Corazza, GR
通讯作者：
Corazza, GR
影响因子：
24.5
作者：
Diosdado, B;Wapenaar, MC;Wijmenga, C
通讯作者：
Wijmenga, C
影响因子：
21.3
作者：
Marcet, Brice;Chevalier, Benoit;Barbry, Pascal
通讯作者：
Barbry, Pascal