EGFR in Tumor-Associated Myeloid Cells Promotes Development of Colorectal Cancer in Mice and Associates With Outcomes of Patients.
EGFR in Tumor-Associated Myeloid Cells Promotes Development of Colorectal Cancer in Mice and Associates With Outcomes of Patients.
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肿瘤相关髓系细胞中的表皮生长因子受体促进小鼠结直肠癌的发展，并与患者的预后相关。
DOI：
10.1053/j.gastro.2017.03.053
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发表时间：
2017-07
期刊：
影响因子：
29.4
通讯作者：
Sibilia M
中科院分区：
文献类型：
--
作者：
Srivatsa S;Paul MC;Cardone C;Holcmann M;Amberg N;Pathria P;Diamanti MA;Linder M;Timelthaler G;Dienes HP;Kenner L;Wrba F;Prager GW;Rose-John S;Eferl R;Liguori G;Botti G;Martinelli E;Greten FR;Ciardiello F;Sibilia M
表皮生长因子受体（EGFR）抑制剂是无RAS突变的转移性结直肠肿瘤患者的一线治疗药物。然而，EGFR抑制剂在这些患者中无效，并且EGFR的肿瘤水平与对治疗的反应无关。我们筛选了人类结直肠肿瘤中EGFR阳性髓样细胞，并研究了它们与患者预后的关系。我们还在小鼠中进行了研究，以评估肿瘤细胞和骨髓细胞中的EGFR表达如何促进结肠炎相关癌症和ApcMin依赖性肠道肿瘤发生的发展。我们对116例结直肠肿瘤活检进行免疫组化和免疫荧光分析，以确定肿瘤和间质中EGFR的水平;我们还收集了肿瘤分期和患者特征及结局的信息。我们使用Mann-Whitney U和Kruskal-Wallis检验将肿瘤EGFR水平与肿瘤分期相关联，并使用Kaplan-Meier方法估计患者的中位生存时间。我们在肠上皮细胞（Villin-Cre; Egfrf/f和Villin-CreERT 2; Egfrf/f小鼠）或骨髓细胞（LysM-Cre; Egfrf/f小鼠）中缺乏EGFR的小鼠中在混合背景下进行实验。这些小鼠与ApcMin/+小鼠一起繁殖;结肠炎相关的癌症和结肠炎分别通过给予葡聚糖硫酸钠（DSS）（有或没有氧化偶氮甲烷（AOM））诱导。通过向小鼠施用他莫昔芬，在发展的肿瘤中激活Villin-CreERT 2。表达全长EGFR的同窝仔用作对照。收集肠组织;通过组织学、免疫组织化学、定量逆转录聚合酶链反应和流式细胞术分析来评估结肠炎的严重程度、肿瘤的数量和大小以及肠屏障的完整性。我们在人结直肠肿瘤间质的髓样细胞中检测到EGFR;髓样细胞EGFR表达与肿瘤转移和较短的患者生存时间相关。在ApcMin/+背景下以及施用AOM和DSS后，从骨髓细胞中缺失EGFR的小鼠形成的肿瘤比相应的EGFR表达对照显著更少和更小。肠上皮细胞EGFR缺失不影响肿瘤生长。此外，在给予AOM和DSS的小鼠中，他莫昔芬诱导的EGFR从已建立的肠肿瘤上皮细胞中的缺失并没有减小肿瘤大小。骨髓细胞中的EGFR信号促进肠肿瘤细胞中STAT 3的活化和Survivin的表达。与对照小鼠或从肠上皮细胞中缺失EGFR的小鼠相比，从骨髓细胞中缺失EGFR的小鼠在DSS给药后发生更严重的结肠炎，其特征在于肠道炎症增加和肠屏障破坏。DSS处理的LysM-Cre; Egfrf/f小鼠结肠中EGFR缺陷型骨髓细胞的白细胞介素6（IL 6）表达减少，与对照组相比，上皮细胞STAT 3活化减少。给予DSS的LysM-Cre; Egfrf/f小鼠的重组IL 6给药保护它们免于体重减轻，并恢复上皮细胞增殖和STAT 3活化，与单独给予DSS的小鼠相比。结直肠肿瘤间质髓样细胞中EGFR表达增加与肿瘤进展和转移性结直肠癌患者生存时间缩短相关。从骨髓细胞而不是肠上皮细胞中删除EGFR可保护小鼠免受结肠炎诱导的肠癌和ApcMin依赖性肠道肿瘤发生。EGFR的髓样细胞表达增加肠上皮细胞中STAT 3的活化和存活素的表达以及结肠组织中IL 6的表达。这些发现表明，EGFR的表达由结直肠肿瘤间质的髓样细胞，而不是癌细胞本身，有助于肿瘤的发展。
Inhibitors of the epidermal growth factor receptor (EGFR) are the first-line therapy for patients with metastatic colorectal tumors without RAS mutations. However, EGFR inhibitors are ineffective in these patients, and tumor level of EGFR does not associate with response to therapy. We screened human colorectal tumors for EGFR-positive myeloid cells and investigated their association with patient outcome. We also performed studies in mice to evaluate how EGFR expression in tumor cells and myeloid cells contributes to development of colitis-associated cancer and ApcMin-dependent intestinal tumorigenesis. We performed immunohistochemical and immunofluorescent analyses of 116 colorectal tumor biopsies to determine levels of EGFR in tumor and stroma; we also collected information on tumor stage and patient features and outcomes. We used the Mann-Whitney U and Kruskal-Wallis tests to correlate tumor levels of EGFR with tumor stage, and the Kaplan-Meier method to estimate patients’ median survival time. We performed experiments in mice lacking EGFR in intestinal epithelial cells (Villin-Cre; Egfrf/f and Villin-CreERT2; Egfrf/f mice) or myeloid cells (LysM-Cre; Egfrf/f mice) on a mixed background. These mice were bred with ApcMin/+ mice; colitis-associated cancer and colitis were induced by administration of dextran sodium sulfate (DSS), with or without azoxymethane (AOM), respectively. Villin-CreERT2 was activated in developed tumors by administration of tamoxifen to mice. Littermates that expressed full-length EGFR were used as controls. Intestinal tissues were collected; severity of colitis, numbers and size of tumors, and intestinal barrier integrity were assessed by histologic, immunohistochemical, quantitative reverse transcription polymerase chain reaction, and flow cytometry analyses. We detected EGFR in myeloid cells in the stroma of human colorectal tumors; myeloid cell expression of EGFR associated with tumor metastasis and shorter patient survival time. Mice with deletion of EGFR from myeloid cells formed significantly fewer and smaller tumors than the respective EGFR-expressing controls in an ApcMin/+ background as well as after administration of AOM and DSS. Deletion of EGFR from intestinal epithelial cells did not affect tumor growth. Furthermore, tamoxifen-induced deletion of EGFR from epithelial cells of established intestinal tumors in mice given AOM and DSS did not reduce tumor size. EGFR signaling in myeloid cells promoted activation of STAT3 and expression of survivin in intestinal tumor cells. Mice with deletion of EGFR from myeloid cells developed more severe colitis after DSS administration, characterized by increased intestinal inflammation and intestinal barrier disruption, than control mice or mice with deletion of EGFR from intestinal epithelial cells. EGFR-deficient myeloid cells in the colon of DSS-treated LysM-Cre; Egfrf/f mice had reduced expression of interleukin 6 (IL6), and epithelial STAT3 activation was reduced compared with controls. Administration of recombinant IL6 to LysM-Cre; Egfrf/f mice given DSS protected them from weight loss and restored epithelial proliferation and STAT3 activation, compared with administration of DSS alone to these mice. Increased expression of EGFR in myeloid cells from the colorectal tumor stroma associates with tumor progression and reduced survival time of patients with metastatic colorectal cancer. Deletion of EGFR from myeloid cells, but not intestinal epithelial cells, protects mice from colitis-induced intestinal cancer and ApcMin-dependent intestinal tumorigenesis. Myeloid cell expression of EGFR increases activation of STAT3 and expression of survivin in intestinal epithelial cells and expression of IL6 in colon tissues. These findings indicate that expression of EGFR by myeloid cells of the colorectal tumor stroma, rather than the cancer cells themselves, contributes to tumor development.
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影响因子：
6.2
作者：
Khorana, AA;Ryan, CK;Sahasrabudhe, DM
通讯作者：
Sahasrabudhe, DM
影响因子：
3
作者：
Clausen, BE;Burkhardt, C;Förster, I
通讯作者：
Förster, I
影响因子：
64.5
作者：
Lichtenberger, Beate M.;Tan, Poi Kiang;Sibilia, Maria
通讯作者：
Sibilia, Maria
影响因子：
--
作者：
Bronte G;Silvestris N;Castiglia M;Galvano A;Passiglia F;Sortino G;Cicero G;Rolfo C;Peeters M;Bazan V;Fanale D;Giordano A;Russo A
通讯作者：
Russo A
影响因子：
2.5
作者：
Lee, D;Cross, SH;Threadgill, DW
通讯作者：
Threadgill, DW