Defects in NADPH Oxidase Genes NOX1 and DUOX2 in Very Early Onset Inflammatory Bowel Disease.
Defects in NADPH Oxidase Genes NOX1 and DUOX2 in Very Early Onset Inflammatory Bowel Disease.
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DOI：
10.1016/j.jcmgh.2015.06.005
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发表时间：
2015-09-01
影响因子：
7.2
通讯作者：
Knaus UG
中科院分区：
文献类型：
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作者：
Hayes P;Dhillon S;O'Neill K;Thoeni C;Hui KY;Elkadri A;Guo CH;Kovacic L;Aviello G;Alvarez LA;Griffiths AM;Snapper SB;Brant SR;Doroshow JH;Silverberg MS;Peter I;McGovern DP;Cho J;Brumell JH;Uhlig HH;Bourke B;Muise AA;Knaus UG
肠道先天防御系统的缺陷使患者易患炎症性肠病（IBD）。由烟酰胺腺嘌呤二核苷酸磷酸（NADPH）氧化酶在粘膜屏障中产生的活性氧（ROS）维持肠道内稳态并抵御病原体的攻击。我们推测IBD患儿可能存在肠道NADPH氧化酶的分子遗传缺陷。对59例极早发性炎症性肠病（VEOIBD）患儿进行上皮细胞NADPH氧化酶NOX 1和DUOX 2的靶向外显子组测序后，使用桑格测序法验证鉴定的突变。通过同源性计算机建模进行N 0X 1和DU 0X 2变体的结构分析。功能表征包括ROS生成模型细胞系和在体内转导的小鼠隐窝，蛋白质表达，细胞内定位，和基于细胞的感染研究与肠道病原体空肠弯曲菌和肠致病性大肠杆菌。我们在209例VEOIBD患者中的5例中发现了NOX 1（c.988G>A，p.Pro330Ser; c.967G>A，p.Asp360Asn）和DUOX 2（c.4474G>A，p.Arg1211Cys; c.3631C>T，p.Arg1492Cys）的错义突变。NOX 1 p.Asp360Asn变异在男性德系犹太溃疡性结肠炎队列中复制。同时具有NOX 1和DUOX 2变异体的患者表现出异常的潘氏细胞化生。与野生型酶相比，所有NOX 1和DUOX 2变体均显示ROS产生减少。尽管适当的细胞定位和相当的病原体刺激的改变氧化酶的易位，细胞窝藏NOX 1或DUOX 2的变体有缺陷的宿主感染空肠弯曲菌的抗性。这项研究确定了与VEOIBD相关的NOX 1和DUOX 2中的第一个失活错义变体。肠上皮细胞产生的ROS缺陷构成了VEOIBD发展的风险因素。
Defects in intestinal innate defense systems predispose patients to inflammatory bowel disease (IBD). Reactive oxygen species (ROS) generated by nicotinamide-adenine dinucleotide phosphate (NADPH) oxidases in the mucosal barrier maintain gut homeostasis and defend against pathogenic attack. We hypothesized that molecular genetic defects in intestinal NADPH oxidases might be present in children with IBD. After targeted exome sequencing of epithelial NADPH oxidases NOX1 and DUOX2 on 59 children with very early onset inflammatory bowel disease (VEOIBD), the identified mutations were validated using Sanger Sequencing. A structural analysis of NOX1 and DUOX2 variants was performed by homology in silico modeling. The functional characterization included ROS generation in model cell lines and in in vivo transduced murine crypts, protein expression, intracellular localization, and cell-based infection studies with the enteric pathogens Campylobacter jejuni and enteropathogenic Escherichia coli. We identified missense mutations in NOX1 (c.988G>A, p.Pro330Ser; c.967G>A, p.Asp360Asn) and DUOX2 (c.4474G>A, p.Arg1211Cys; c.3631C>T, p.Arg1492Cys) in 5 of 209 VEOIBD patients. The NOX1 p.Asp360Asn variant was replicated in a male Ashkenazi Jewish ulcerative colitis cohort. Patients with both NOX1 and DUOX2 variants showed abnormal Paneth cell metaplasia. All NOX1 and DUOX2 variants showed reduced ROS production compared with wild-type enzymes. Despite appropriate cellular localization and comparable pathogen-stimulated translocation of altered oxidases, cells harboring NOX1 or DUOX2 variants had defective host resistance to infection with C. jejuni. This study identifies the first inactivating missense variants in NOX1 and DUOX2 associated with VEOIBD. Defective ROS production from intestinal epithelial cells constitutes a risk factor for developing VEOIBD.