Immune Cell Production of Interleukin 17 Induces Stem Cell Features of Pancreatic Intraepithelial Neoplasia Cells.
Immune Cell Production of Interleukin 17 Induces Stem Cell Features of Pancreatic Intraepithelial Neoplasia Cells.
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DOI：
10.1053/j.gastro.2018.03.041
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发表时间：
2018-07
期刊：
影响因子：
29.4
通讯作者：
McAllister F
中科院分区：
文献类型：
--
作者：
Zhang Y;Zoltan M;Riquelme E;Xu H;Sahin I;Castro-Pando S;Montiel MF;Chang K;Jiang Z;Ling J;Gupta S;Horne W;Pruski M;Wang H;Sun SC;Lozano G;Chiao P;Maitra A;Leach SD;Kolls JK;Vilar E;Wang TC;Bailey JM;McAllister F
人们对免疫系统如何影响胰腺癌细胞的干细胞特征知之甚少。在慢性炎症胰腺（慢性胰腺炎）中产生白细胞介素17 A（IL 17 A）的免疫细胞有助于PanIN的启动和进展。我们研究了IL 17 A信号传导对胰腺癌祖细胞的影响以及这种现象的临床相关性。我们对Mist 1Cre、LSLKras、Rosa 26 mTmG（KCiMist;G）和Kras（G12 D）、Trp 53（R172 H）、Pdx 1-Cre（KPC）小鼠（在他莫昔芬诱导后自发发生胰腺上皮内瘤形成，PanIN）和对照同窝小鼠进行了研究。给一些小鼠注射针对IL 17 A的中和抗体或对照抗体。收集胰腺，通过基于谱系追踪的流式细胞术分离PanIN上皮细胞，并比较基因表达谱。我们从KPC小鼠的胰腺肿瘤中收集细胞，将其与IL 17或对照培养基一起孵育，测量由IL 17信号转导调节的基因的表达，将癌细胞注射到免疫活性小鼠中，并测量肿瘤生长。IL 17 A在KCiMist小鼠的胰腺中从腺病毒载体过表达。从所有小鼠收集胰腺并通过组织学和免疫组织化学分析。使用小干扰或小发夹RNA敲低KPC胰腺癌细胞中的双皮质素样激酶1（DCLK 1）和其他蛋白质的水平;通过免疫印迹分析细胞。我们从患者中获得了65个胰腺肿瘤标本，通过免疫组化分析蛋白水平，并将结果与患者生存时间进行比较。我们还使用癌症基因组图谱数据库分析了基因表达水平和患者结局。来自KCiMist;G小鼠的PanIN细胞具有与胚胎干细胞相关的基因表达模式。注射IL 17中和抗体或不分泌IL 17的免疫细胞的小鼠失去了这种表达模式，并显著降低了调节簇细胞发育的DCLK 1和POU 2类同源框3（POU 2F 3）的表达。与对照小鼠相比，过表达IL 17的KCiMist小鼠形成了更多的PanIN，其中DCLK 1阳性细胞更多。与对照培养基中的细胞相比，暴露于IL 17 A的来自KPC小鼠的胰腺肿瘤细胞和人Capan-2细胞具有增加的NF-κB和MAPK信号传导的活化，以及增加的DCLK 1和ALDH 1A 1（胚胎干细胞的标志物）表达。当这些细胞被注射到免疫活性小鼠中时，它们也比未暴露于IL 17的细胞更快地形成肿瘤。与未敲除的细胞相比，敲除DCLK 1的KPC细胞在与IL 17孵育后表达较低水平的ALDH 1A 1。与DCLK 1阴性PanIN细胞相比，来自小鼠的DCLK 1阳性PanIN细胞中IL 17受体C（IL 17 RC）的表达更高。在人胰腺肿瘤组织中，高水平的DCLK 1与患者的中位生存时间较短相关（17.7个月，而肿瘤中DCLK 1水平较低的患者为26.6个月）。POU 2F 3和LAMC 2的肿瘤水平也与患者的生存时间相关。在小鼠和人胰腺肿瘤和前体的研究中，我们发现免疫细胞衍生的IL 17通过增加DCLK 1、POU 2F 3、ALDH 1A 1和IL 17 RC的表达来调节胰腺癌细胞的簇细胞和干细胞特征的发育。破坏这一通路的策略可能会被开发出来，以防止胰腺肿瘤的生长和进展。
Little is known about how the immune system affects stem cell features of pancreatic cancer cells. Immune cells that produce interleukin 17 A (IL17A) in the chronically inflamed pancreas (chronic pancreatitis) contribute to PanIN initiation and progression. We investigated the effects of IL17A signaling exerts on pancreatic cancer progenitor cells and the clinical relevance of this phenomena. We performed studies with Mist1Cre;LSLKras;Rosa26mTmG (KCiMist;G) and Kras(G12D); Trp53(R172H); Pdx1-Cre (KPC) mice (which upon tamoxifen induction spontaneously develop pancreatic intraepithelial neoplasias, PanINs) and control littermates. Some mice were injected with neutralizing antibodies against IL17A or control antibody. Pancreata were collected, PanIN epithelial cells were isolated by flow cytometry based on lineage tracing, and gene expression profiles were compared. We collected cells from pancreatic tumors of KPC mice, incubated them with IL17 or control media, measured expression of genes regulated by IL17 signaling, injected the cancer cells into immune competent mice, and measured tumor growth. IL17A was overexpressed in pancreata of KCiMist mice from an adenoviral vector. Pancreata were collected from all mice and analyzed by histology and immunohistochemistry. Levels of doublecortin like kinase 1 (DCLK1) and other proteins were knocked down in KPC pancreatic cancer cells using small interfering or small hairpin RNAs; cells were analyzed by immunoblotting. We obtained 65 pancreatic tumor specimens from patients, analyzed protein levels by immunohistochemistry, and compared results with patient survival times. We also analyzed gene expression levels and patient outcome using the Cancer Genome Atlas database. PanIN cells from KCiMist;G mice had a gene expression pattern associated with embryonic stem cells. Mice given injections of IL17 neutralizing antibodies, or with immune cells that did not secrete IL17, lost this expression pattern, and significantly decreased expression of DCLK1 and POU class 2 homeobox 3 (POU2F3), which regulate tuft cell development. KCiMist mice that overexpressed IL17 formed more PanINs, with more DCLK1-positive cells, than control mice. Pancreatic tumor cells from KPC mice and human Capan-2 cells exposed to IL17A had increased activation of NF-κB and MAPK signaling, and increased expression of DCLK1 and ALDH1A1 (a marker of embryonic stem cells), compared to cells in control media. These cells also formed tumors faster that cells not exposed to IL17 when they were injected into immunocompetent mice. KPC cells with knockdown of DCLK1 expressed lower levels of ALDH1A1 following incubation with IL17 than cells without knockdown. Expression of the IL17 receptor C (IL17RC) was higher in DCLK1-positive PanIN cells from mice compared to DCLK1-negative PanIN cells. In human pancreatic tumor tissues, high levels of DCLK1 associated with a shorter median survival time of patients (17.7 months, compared with 26.6 months of patients whose tumors had low levels of DCLK1). Tumor levels of POU2F3 and LAMC2 also associated with patient survival time. In studies of mouse and human pancreatic tumors and precursors, we found immune cell-derived IL17 to regulate development of tuft cells and stem cell features of pancreatic cancer cells via increased expression of DCLK1, POU2F3, ALDH1A1, and IL17RC. Strategies to disrupt this pathway might be developed to prevent pancreatic tumor growth and progression.
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影响因子：
11.2
作者：
Benevides L;da Fonseca DM;Donate PB;Tiezzi DG;De Carvalho DD;de Andrade JM;Martins GA;Silva JS
通讯作者：
Silva JS
影响因子：
14.9
作者：
Colaprico A;Silva TC;Olsen C;Garofano L;Cava C;Garolini D;Sabedot TS;Malta TM;Pagnotta SM;Castiglioni I;Ceccarelli M;Bontempi G;Noushmehr H
通讯作者：
Noushmehr H
影响因子：
64.8
作者：
Gerbe F;Sidot E;Smyth DJ;Ohmoto M;Matsumoto I;Dardalhon V;Cesses P;Garnier L;Pouzolles M;Brulin B;Bruschi M;Harcus Y;Zimmermann VS;Taylor N;Maizels RM;Jay P
通讯作者：
Jay P
影响因子：
2.6
作者：
Fedchenko N;Reifenrath J
通讯作者：
Reifenrath J
影响因子：
4.6
作者：
Bie Q;Sun C;Gong A;Li C;Su Z;Zheng D;Ji X;Wu Y;Guo Q;Wang S;Xu H
通讯作者：
Xu H