SIGIRR Mutation in Human Necrotizing Enterocolitis (NEC) Disrupts STAT3-Dependent microRNA Expression in Neonatal Gut.
SIGIRR Mutation in Human Necrotizing Enterocolitis (NEC) Disrupts STAT3-Dependent microRNA Expression in Neonatal Gut.
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DOI：
10.1016/j.jcmgh.2021.09.009
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发表时间：
2022
影响因子：
7.2
通讯作者：
Sampath V
中科院分区：
文献类型：
--
作者：
Yu W;Haque I;Venkatraman A;Menden HL;Mabry SM;Roy BC;Xia S;Prokop JW;Umar S;Geurts AM;Sampath V
单免疫球蛋白白介素-1相关受体（SIGIRR）是toll样受体信号传导的主要抑制剂。我们的实验室在一名死于严重坏死性小肠结肠炎（NEC）的婴儿中发现了一种新的SIGIRR停止突变（p.Y168X）。在此，我们研究了SIGIRR突变诱导新生儿肠道toll样受体高反应性，破坏出生后肠道适应的机制。聚类规则间隔短回文重复序列(CRISPR)/Cas9用于产生编码SIGIRR p.Y168X突变的转基因小鼠。回肠裂解物、小鼠肠上皮细胞（IEC）裂解物和肠切片用于评估炎症、信号传导和转录激活物3 （STAT3）磷酸化、microRNA （miRNA）和白细胞介素-1相关激酶1 （IRAK1）的表达。通过Western blot、定量逆转录聚合酶链反应（qRT-PCR）和荧光素酶检测，研究了SIGIRR - stat3信号在表达野生型或SIGIRR （p.Y168X）质粒的人肠上皮细胞（HIEC）中的表达。SigirrTg小鼠表现出肠道炎症和核因子κ b活化增加，同时miR-146a和miR-155的IEC表达降低。HIECs中的机制研究表明，尽管SIGIRR诱导了stat3介导的miR-146a和miR-155的表达，但p.Y168X突变破坏了SIGIRR介导的stat3依赖性miRNA的表达。染色质免疫沉淀和荧光素酶分析显示，stat3诱导的miRNA表达的SIGIRR激活依赖于IRAK1。在hiec和小鼠肠道中，通过p.Y168X突变观察到miR-146a表达降低，IRAK1表达增加，IRAK1是一种蛋白质，其下调对出生后肠道适应很重要。我们的研究结果揭示了SIGIRR调节出生后肠道适应的新途径（SIGIRR - stat3 - mirna - irak1抑制），该途径被人类NEC中发现的SIGIRR突变破坏。这些数据为NEC中SIGIRR基因突变如何导致出生后肠道免疫耐受丧失提供了新的见解。
Single immunoglobulin interleukin-1–related receptor (SIGIRR) is a major inhibitor of Toll-like receptor signaling. Our laboratory identified a novel SIGIRR stop mutation (p.Y168X) in an infant who died of severe necrotizing enterocolitis (NEC). Herein, we investigated the mechanisms by which SIGIRR mutations induce Toll-like receptor hyper-responsiveness in the neonatal gut, disrupting postnatal intestinal adaptation. Clustered regularly interspaced short palindromic repeats (CRISPR)/Cas9 was used to generate transgenic mice encoding the SIGIRR p.Y168X mutation. Ileal lysates, mouse intestinal epithelial cell (IEC) lysates, and intestinal sections were used to assess inflammation, signal transducer and activator of transcription 3 (STAT3) phosphorylation, microRNA (miRNA), and interleukin-1–related–associated kinase 1 (IRAK1) expression. Western blot, quantitative reverse-transcription polymerase chain reaction(qRT-PCR), and luciferase assays were performed to investigate SIGIRR–STAT3 signaling in human intestinal epithelial cells (HIEC) expressing wild-type or SIGIRR (p.Y168X) plasmids. SigirrTg mice showed increased intestinal inflammation and nuclear factor-κB activation concomitant with decreased IEC expression of miR-146a and miR-155. Mechanistic studies in HIECs showed that although SIGIRR induced STAT3-mediated expression of miR-146a and miR-155, the p.Y168X mutation disrupted SIGIRR-mediated STAT3-dependent miRNA expression. Chromatin immunoprecipitation and luciferase assays showed that SIGIRR activation of STAT3-induced miRNA expression is dependent on IRAK1. Both in HIECs and in the mouse intestine, decreased expression of miR-146a observed with the p.Y168X mutation increased expression of IRAK1, a protein whose down-regulation is important for postnatal gut adaptation. Our results uncover a novel pathway (SIGIRR–STAT3–miRNA–IRAK1 repression) by which SIGIRR regulates postnatal intestine adaptation, which is disrupted by a SIGIRR mutation identified in human NEC. These data provide new insights into how human genetic mutations in SIGIRR identified in NEC result in loss of postnatal intestinal immune tolerance.
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DOI：
10.1073/pnas.0401710101
发表时间：
2004-05-11
影响因子：
11.1
作者：
Claud, EC;Lu, L;Cherayil, BJ
通讯作者：
Cherayil, BJ
影响因子：
3.7
作者：
Nanthakumar N;Meng D;Goldstein AM;Zhu W;Lu L;Uauy R;Llanos A;Claud EC;Walker WA
通讯作者：
Walker WA
影响因子：
2.9
作者：
Krieger, E;Koraimann, G;Vriend, G
通讯作者：
Vriend, G
影响因子：
4.4
作者：
Leaphart, Cynthia L.;Cavallo, Jaime;Hackam, David J.
通讯作者：
Hackam, David J.
DOI：
10.4049/jimmunol.1202264
发表时间：
2013-04-01
期刊：
Journal of immunology (Baltimore, Md. : 1950)
影响因子：
--
作者：
Neal MD;Sodhi CP;Dyer M;Craig BT;Good M;Jia H;Yazji I;Afrazi A;Richardson WM;Beer-Stolz D;Ma C;Prindle T;Grant Z;Branca MF;Ozolek J;Hackam DJ
通讯作者：
Hackam DJ