Fucosylation Deficiency in Mice Leads to Colitis and Adenocarcinoma.
Fucosylation Deficiency in Mice Leads to Colitis and Adenocarcinoma.
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DOI：
10.1053/j.gastro.2016.09.004
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发表时间：
2017-01
期刊：
影响因子：
29.4
通讯作者：
Zhou L
中科院分区：
文献类型：
--
作者：
Wang Y;Huang D;Chen KY;Cui M;Wang W;Huang X;Awadellah A;Li Q;Friedman A;Xin WW;Di Martino L;Cominelli F;Miron A;Chan R;Fox JG;Xu Y;Shen X;Kalady MF;Markowitz S;Maillard I;Lowe JB;Xin W;Zhou L
岩藻糖底物GDP-岩藻糖的从头合成需要由GDP-甘露糖4，6-脱水酶(GMDS)和GDP-4-酮-6-脱氧甘露糖-3，5-差向异构酶-4-还原酶(FX或TSTA3)介导的一系列反应。在6%-13%的结直肠癌中发现GMDS缺失和突变；这些主要影响升结肠和横结肠。我们调查了GDP-岩藻糖合成缺失导致的岩藻糖基化缺失是否与结肠癌的发生有关。FX缺乏症和GMDS缺失症的生化表型与GDP-岩藻糖缺乏症相同。我们研究了岩藻糖基化缺陷小鼠模型(FX-/-小鼠)和携带全长FX基因的小鼠(对照组)。将小鼠置于标准食物或含岩藻糖的饮食中(相当于对照岩藻糖聚糖表型)。收集结肠组织并进行组织学或ELISA分析，以测量细胞因子水平；也收集T细胞并进行分析。粪便标本进行16S rRNA测序分析。通过摄取荧光葡聚糖来检测粘膜屏障功能。我们将FX-/-或对照小鼠(Ly5.2)的骨髓细胞移植到8周龄FX-/-或对照小鼠(Ly5.1)中。我们对Notch和HES家族转录因子(HES1)在小鼠结肠组织和60例人类结直肠癌标本(左侧27例，右侧33例)中的表达进行了免疫组织化学分析。FX-/-小鼠出现结肠炎和锯齿状病变。在饮食中添加岩藻糖，通过挽救途径恢复岩藻糖基化，逆转了FX-/-小鼠的肠道病理。在没有岩藻糖基化的情况下，高达40%的小鼠出现不典型增生并进展为腺癌，主要影响到右半结肠和盲肠。在饲喂标准饲料的FX-/-小鼠中，Notch不被激活，导致其靶标Hes1的表达减少。岩藻糖基化缺陷改变了粪便微生物区系的组成，降低了粘膜屏障功能，并改变了以Ki67为标志的上皮细胞增殖。接受对照骨髓细胞的FX-/-小鼠出现肠道炎症和异型增生，细胞毒性T细胞产生的细胞因子表达减少。与其他类型的大肠肿瘤或非肿瘤组织相比，具有MLH1表观遗传缺失的人无柄锯齿状腺瘤和右侧大肠肿瘤已丢失或具有较低水平的HES1。在小鼠中，岩藻糖基化缺陷导致结肠炎和腺癌，Notch活性丧失，Hes1下调。HES1缺失与人类右侧结直肠肿瘤的发生和MLH1的表观遗传缺失相关。这些发现表明，结肠癌的一个亚组的癌变是由岩藻糖基化缺陷和/或HES1-缺失驱动的分子机制的结果。
De novo synthesis of GDP-fucose, a substrate for fucosylglycans, requires sequential reactions mediated by GDP-mannose 4,6-dehydratase (GMDS) and GDP-4-keto-6-deoxymannose 3,5-epimerase-4-reductase (FX or TSTA3). GMDS deletions and mutations are found in 6%–13% of colorectal cancers; these mostly affect ascending and transverse colon. We investigated whether lack of fucosylation consequent to loss of GDP-fucose synthesis contributes to colon carcinogenesis. FX deficiency and GMDS deletion produce the same biochemical phenotype of GDP-fucose deficiency. We studied a mouse model of fucosylation deficiency (Fx–/– mice) and mice with the full-length Fx gene (controls). Mice were placed on standard chow or fucose-containing diet (equivalent to a control fucosylglycan phenotype). Colon tissues were collected and analyzed histologically or by ELISAs to measure cytokine levels; T cells were also collected and analyzed. Fecal samples were analyzed by 16s rRNA sequencing. Mucosal barrier function was measured by uptake of fluorescent dextran. We transplanted bone marrow cells from Fx–/– or control mice (Ly5.2) into irradiated 8-week old Fx–/– or control mice (Ly5.1). We performed immunohistochemical analyses for expression of Notch and the hes family bHLH transcription factor (HES1) in colon tissues from mice and a panel of 60 human colorectal cancer specimens (27 left-sided, 33 right-sided). Fx–/– mice developed colitis and serrated-like lesions. The intestinal pathology of Fx–/– mice was reversed by addition of fucose to the diet, which restored fucosylation via a salvage pathway. In the absence of fucosylation, dysplasia appeared and progressed to adenocarcinoma in up to 40% of mice, affecting mainly the right colon and cecum. Notch was not activated in Fx–/– mice fed standard chow, leading to decreased expression of its target Hes1. Fucosylation deficiency altered the composition of the fecal microbiota, reduced mucosal barrier function and altered epithelial proliferation marked by Ki67. Fx–/– mice receiving control bone marrow cells had intestinal inflammation and dysplasia, and reduced expression of cytokines produced by cytotoxic T cells. Human sessile serrated adenomas and right-sided colorectal tumors with epigenetic loss of MLH1 had lost or had lower levels of HES1 than other colorectal tumor types or nontumor tissues. In mice, fucosylation deficiency leads to colitis and adenocarcinoma, loss of Notch activation, and downregulation of Hes1. HES1 loss correlates with development of human right-sided colorectal tumors with epigenetic loss of MLH1. These findings indicate that carcinogenesis in a subset of colon cancer is consequent to a molecular mechanism driven by fucosylation deficiency and/or HES1-loss.
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影响因子：
3
作者：
Langfelder P;Horvath S
通讯作者：
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影响因子：
5.3
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发表时间：
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影响因子：
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作者：
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DOI：
10.1097/pas.0000000000000531
发表时间：
2016-01
期刊：
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影响因子：
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作者：
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