Transplantable programmed death ligand 1 expressing gastroids from gastric cancer prone Nfkb1(-/-) mice.
Transplantable programmed death ligand 1 expressing gastroids from gastric cancer prone Nfkb1(-/-) mice.
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DOI：
10.1038/s41419-021-04376-2
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发表时间：
2021-11-17
影响因子：
9
通讯作者：
O'Reilly LA
中科院分区：
文献类型：
--
作者：
Low JT;Ho GY;Scott M;Tan CW;Whitehead L;Barber K;Yip HYK;Dekkers JF;Hirokawa Y;Silke J;Burgess AW;Strasser A;Putoczki TL;O'Reilly LA
胃癌(GC)是全球第五大常见癌症和第三大癌症相关死亡原因[1]。GC有几种组织学亚型[2]，肠型GC(IGC)最为常见。IGC是由炎性胃炎引起的，通常由幽门螺杆菌或EBV感染引起。这导致了NF-κB转录因子的持续激活，从而驱动了被认为促进肿瘤形成的炎症因子的表达[2]。程序性死亡配体1(PD-L1)是表达在T细胞上的免疫检查点调节因子PD-1的配体。PD-L1/PD1相互作用抑制细胞毒性T细胞介导的对癌细胞的杀伤[3]。肿瘤可以劫持这一途径，例如通过表达PD-L1，使它们对这种免疫攻击产生抵抗[3]。免疫检查点抑制疗法(ICIT)通过阻断PD1或CTLA4或PD1配体PDL1/PDL2来增强对包括GC[4]在内的恶性肿瘤细胞的杀伤。我们建立了一种由转录因子Rel/NF-κB家族成员之一的NF-κB1缺失驱动的IGC鼠模型。Nfkb1−/−小鼠出现胃部肿瘤坏死因子表达异常增加和STAT1激活，导致炎症免疫反应，最终发展为GC[5，6]。肿瘤前期Nfkb1−/−小鼠的胃内细胞表现出异常的增殖、JAK/STAT1信号和PD-L1表达[5，6](补充图1A)。与之相关的是，人类NFKB1基因中降低其功能的多态与胃癌风险的增加有关[6]。人类GC的EBV+和微卫星不稳定性亚型[7]表现出与Nfkb1−/−小鼠的GC相似的特征，这表明它们也是由持续的炎症和免疫激活驱动的，它们可能从ICIT[6]中受益。来自人类GC组织和动物模型的有机化合物是GC研究和测试新的靶向疗法的重要实验工具[8]。我们从年轻、健康和年长的携带Nfkb1肿瘤的−/−小鼠的胃中产生了胃有机化合物(GO)(图1A，补充1B，补充表1，2)。从幼年(7-8周)Wt或Nfkb1−/−的胃细胞中提取的胃器官(GO)呈球状、囊状和萌芽状(图1A，B，补充2)，如前所述[9]。定量明场显微镜显示，在培养的第4、6或8天，Nfkb1−/−和对照wtGO之间的有机类化合物总数、萌芽总数(总数)和萌发潜力(频率)没有显著差异(图1C-F)。我们还从Nfkb1−/−小鼠的胃肿瘤(TGO)中提取了有机类化合物。从这些小鼠(17月龄，1391号，1392号，1399号)采集胃肿瘤组织，进行组织学检查、流式细胞仪分析和有机物培养。这些小鼠的组织学检查证实了异型增生和肿瘤细胞的胃侵袭(图1G)。与我们在肿瘤前Nfkb1−/−小鼠[5，6]中的发现一致，与wt对照组小鼠相比，胃中髓系和上皮细胞上PD-L1蛋白的异常表达，流式细胞仪分析显示所有GC负荷的Nfkb1−/−小鼠胃中CD11b+髓系细胞和Epcam+上皮细胞上的PD-L1水平都很高(补充图3A)。从每个肿瘤样本中产生TGO，并如上所述进行扩展(图1H，补充表1，2，补充方法)。所有TGO均呈椭圆形、囊状或芽状(图1H)，可扩增至少30代(~60…
Gastric cancer (GC) is the fifth most common cancer and the third highest cause of cancer-related deaths globally [1]. There are several histological subtypes of GC [2], with intestinal type GC (IGC) the most common. IGC is initiated by inflammatory gastritis, often driven by Helicobacter pylori (H. pylori) or EBV infection [1]. This results in sustained activation of NF-κB transcription factors, which drive the expression of inflammatory factors thought to promote tumorigenesis [2]. Programmed death ligand 1 (PD-L1) is the ligand of the immune checkpoint regulator PD-1 that is expressed on T cells. The PD-L1/PD1 interaction inhibits cytotoxic T cell mediated killing of cancer cells [3]. Tumours can hijack this pathway, for example by expressing PD-L1, to render them resistant to such immune attack [3]. Immune checkpoint inhibitor therapy (ICIT) enhances the killing of malignant cells, including GC [4], by blocking PD1 or CTLA4 or the PD1 ligands, PDL1/PDL2. We developed a mouse model of IGC that is driven by loss of NF-κB1, a member of the REL/NF-κB family of transcription factors. Nfkb1−/− mice present with abnormally increased expression of TNF and activation of STAT1 in the stomach, resulting in an inflammatory immune response that culminates in the development of GC [5, 6]. Cells within the stomachs from pre-neoplastic Nfkb1−/− mice display abnormally increased proliferation, JAK/STAT1 signalling and PD-L1 expression [5, 6](Supplementary Fig. 1A). Pertinently, polymorphisms in human NFKB1 that diminish its function have been linked with increased risk for GC [6]. The EBV+ and Microsatellite Instabilityhi (MSIhi) subtypes of human GC [7] exhibit features similar to the GC that arise in Nfkb1−/− mice, suggesting that they are also driven by sustained inflammation, immune activation and that they may benefit from ICIT [6]. Organoids derived from human GC tissue and from animal models are an important experimental tool in GC research and for the testing of novel targeted therapies [8]. We generated gastric organoids (GOs) from the stomachs of both young, healthy and older tumour bearing Nfkb1−/− mice (Fig. 1 A, Supplementary 1B, Supplementary Tables 1, 2). Gastric organoids (GOs) derived from cells from the stomachs of young (7–8 week) wt or Nfkb1−/− exhibited spheroid-like, cyst-like and budding morphologies (Fig. 1 A, B, Supplementary 2) as previously described [9]. Quantitative bright field microscopy revealed that there were no significant differences between the total numbers of organoids, budding (total numbers) or budding potential (frequencies) between Nfkb1−/− and control wt GOs on days 4, 6 or 8 of culture (Fig. 1 C–F).We also derived organoids from gastric tumours (TGOs) of Nfkb1−/− mice. Gastric tumour tissue was harvested from these mice (> 17 months,# 1391, 1392, 1399) for histological examination, flow cytometric analysis and organoid culture. Histological examination of these mice confirmed dysplasia and gastric invasion of neoplastic cells (Fig. 1 G). Consistent with our findings in pre-neoplastic Nfkb1−/− mice [5, 6], which showed abnormally elevated PD-L1 protein expression on myeloid and epithelial cells in the stomach compared to wt control mice, flow cytometric analysis revealed high levels of PD-L1 on both CD11b+ myeloid cells and Epcam+ epithelial cells in the stomachs of all GC-burdened Nfkb1−/− mice (Supplementary Fig. 3A). TGOs were generated from each tumour sample and expanded as described above (Fig. 1 H, Supplementary Tables 1, 2, Supplementary methods). All TGOs exhibited either spheroid, cystic or budding morphology (Fig. 1 H) and could be expanded for at least 30 passages (~ 60 …
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DOI：
10.1038/nrc3239
发表时间：
2012-03-22
期刊：
Nature reviews. Cancer
影响因子：
--
作者：
Pardoll DM
通讯作者：
Pardoll DM
DOI：
10.1126/science.aao3130
发表时间：
2017-10-13
期刊：
Science (New York, N.Y.)
影响因子：
--
作者：
Drost J;van Boxtel R;Blokzijl F;Mizutani T;Sasaki N;Sasselli V;de Ligt J;Behjati S;Grolleman JE;van Wezel T;Nik-Zainal S;Kuiper RP;Cuppen E;Clevers H
通讯作者：
Clevers H
影响因子：
29.4
作者：
Low, Jun T.;Christie, Michael;O'Reilly, Lorraine A.
通讯作者：
O'Reilly, Lorraine A.
影响因子：
4.6
作者：
Busuttil RA;Liu DS;Di Costanzo N;Schröder J;Mitchell C;Boussioutas A
通讯作者：
Boussioutas A
影响因子：
64.5
作者：
Biton M;Haber AL;Rogel N;Burgin G;Beyaz S;Schnell A;Ashenberg O;Su CW;Smillie C;Shekhar K;Chen Z;Wu C;Ordovas-Montanes J;Alvarez D;Herbst RH;Zhang M;Tirosh I;Dionne D;Nguyen LT;Xifaras ME;Shalek AK;von Andrian UH;Graham DB;Rozenblatt-Rosen O;Shi HN;Kuchroo V;Yilmaz OH;Regev A;Xavier RJ
通讯作者：
Xavier RJ