Interleukin 1 beta and Matrix Metallopeptidase 3 Contribute to Development of Epidermal Growth Factor Receptor-Dependent Serrated Polyps in Mouse Cecum.
Interleukin 1 beta and Matrix Metallopeptidase 3 Contribute to Development of Epidermal Growth Factor Receptor-Dependent Serrated Polyps in Mouse Cecum.
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白细胞介素 1 β 和基质金属肽酶 3 有助于小鼠盲肠中表皮生长因子受体依赖性锯齿状息肉的形成。
DOI：
10.1053/j.gastro.2019.08.025
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发表时间：
2019
期刊：
影响因子：
29.4
通讯作者：
Lira,SergioA
中科院分区：
文献类型：
--
作者：
He,Zhengxiang;Chen,Lili;Chen,Grace;Smaldini,Paola;Bongers,Gerold;Catalan-Dibene,Jovani;Furtado,GlauciaC;Lira,SergioA
背景与目的在肠上皮细胞中表达表皮生长因子受体（EGFR）配体HBEGF（肝素结合表皮生长因子样生长因子）和组成型活性G蛋白偶联受体（US28）的转基因小鼠（HBUS）在盲肠中产生锯齿状息肉。锯齿状息肉的发展取决于肠道微生物群的组成，并与固有层的细菌入侵有关，伴随着炎症的诱导和盲肠中白细胞介素1 β（IL1B）和基质金属蛋白酶（MMP）3的上调。我们调查的机制，这些变化有助于锯齿状polyps.MethodsWe的发展与C57 BL/6（对照）和HBUS小鼠进行了研究。为了加速息肉的发展，我们通过注射白喉毒素增加了细菌对固有层的暴露，白喉毒素结合上皮细胞表达的转基因HBEGF并引起细胞凋亡。给小鼠注射IL1B中和抗体和MMP激动剂或N-异丁基-N-（4-甲氧基苯磺酰基）甘氨酰异羟肟酸。从小鼠收集肠组织，并通过组织学、逆转录聚合酶链反应、酶联免疫吸附测定、免疫荧光和流式细胞术进行分析。我们使用单细胞RNA测序检查了息肉中的成纤维细胞亚群。结果对HBUS小鼠给予白喉毒素加速了锯齿状息肉的发育（95%的治疗小鼠在100天之前出现息肉，而给予载体的小鼠这一比例为53%）。IL 1B刺激从盲肠分离的血小板衍生生长因子受体α +（PDGRFA+）成纤维细胞亚群，导致MMP 3的表达增加。针对IL1B的中和抗体或MMP抑制剂的施用减少了HBUS小鼠中形成的锯齿状息肉的数量。单细胞RNA测序分析表明，锯齿状息肉中的成纤维细胞的亚群，表达基因，调节基质成纤维细胞和inflammation.ConclusionsIn小鼠的研究，我们发现，屏障破坏和表达的炎症因子有助于锯齿状息肉的发展。盲肠PDGFRA+成纤维细胞的亚群在锯齿状息肉发育的早期阶段通过从骨髓细胞释放IL 1B而被激活。由PDGFRA+成纤维细胞产生的MMP 3对锯齿状息肉的发育很重要。我们的研究结果证实了以前确定的锯齿状息肉相关分子的功能，并表明免疫和基质细胞在锯齿状息肉发育中的作用。
Background & AimsTransgenic mice (HBUS) that express the epidermal growth factor receptor (EGFR) ligand HBEGF (heparin-binding epidermal growth factor–like growth factor) and a constitutively active G protein–coupled receptor (US28) in intestinal epithelial cells develop serrated polyps in the cecum. Development of serrated polyps depends on the composition of the gut microbiota and is associated with bacterial invasion of the lamina propria, accompanied by induction of inflammation and up-regulation of interleukin 1 beta (IL1B) and matrix metalloproteinase (MMP) 3 in the cecum. We investigated the mechanisms by which these changes contribute to development of serrated polyps.MethodsWe performed studies with C57BL/6 (control) and HBUS mice. To accelerate polyp development, we increased the exposure of the bacteria to the lamina propria by injecting HBUS mice with diphtheria toxin, which binds transgenic HBEGF expressed by the epithelial cells and causes apoptosis. Mice were given injections of IL1B-neutralizing antibody and the MMP inhibitorN-isobutyl-N-(4-methoxyphenylsulfonyl)glycyl hydroxamic acid. Intestinal tissues were collected from mice and analyzed by histology, reverse-transcription polymerase chain reaction, enzyme-linked immunosorbent assay, immunofluorescence, and flow cytometry. We examined fibroblast subsets in polyps using single-cell RNA sequencing.ResultsAdministration of diphtheria toxin to HBUS mice accelerated development of serrated polyps (95% of treated mice developed polyps before 100 days of age, compared with 53% given vehicle). IL1B stimulated subsets of platelet-derived growth factor receptor alpha+(PDGRFA+) fibroblasts isolated from cecum, resulting in increased expression of MMP3. Neutralizing antibodies against IL1B or administration of the MMP inhibitor reduced the number of serrated polyps that formed in the HBUS mice. Single-cell RNA sequencing analysis showed subsets of fibroblasts in serrated polyps that express genes that regulate matrix fibroblasts and inflammation.ConclusionsIn studies of mice, we found that barrier breakdown and expression of inflammatory factors contribute to development of serrated polyps. Subsets of cecal PDGFRA+fibroblasts are activated by release of IL1B from myeloid cells during the early stages of serrated polyp development. MMP3 produced by PDGFRA+fibroblasts is important for serrated polyp development. Our findings confirm the functions of previously identified serrated polyp–associated molecules and indicate roles for immune and stromal cells in serrated polyp development.