Inhibition of Aurora Kinase A Induces Necroptosis in Pancreatic Carcinoma.
Inhibition of Aurora Kinase A Induces Necroptosis in Pancreatic Carcinoma.
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抑制 Aurora 激酶 A 诱导胰腺癌坏死性凋亡
DOI：
10.1053/j.gastro.2017.07.036
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发表时间：
2017-11
期刊：
影响因子：
29.4
通讯作者：
Tang D
中科院分区：
文献类型：
--
作者：
Xie Y;Zhu S;Zhong M;Yang M;Sun X;Liu J;Kroemer G;Lotze M;Zeh HJ 3rd;Kang R;Tang D
诱导非凋亡性细胞死亡可能是消除抗凋亡肿瘤的一种方法。我们在胰腺导管腺癌（PDAC）小鼠模型中研究了基于坏死性凋亡的治疗。我们筛选了273种市售激酶抑制剂对人PDAC细胞系（PANC 1）的细胞毒性。我们评估了极光激酶抑制剂CCT 137690刺激PDAC细胞系（PANC 1、PANC 2.03、CFPAC 1、MiaPaCa 2、BxPc 3和PANC 02）和HEK 293细胞系中坏死性凋亡的能力，测量了质膜完整性的丧失、细胞体积的增加、肿胀的细胞器和细胞质空泡。我们测试了CCT 137690在结肠形成试验中的作用，以及坏死性凋亡（坏死抑素-1和坏死磺酰胺）、细胞凋亡、自噬和铁凋亡抑制剂的作用。我们从Pdx 1-Cre; K-RasG 12 D/+; p53 R172 H/+（KPC）小鼠中产生的肿瘤中获得细胞。细胞死亡途径中编码蛋白质的基因被敲除、敲低或从PDAC细胞系中的转基因表达。无胸腺裸鼠或B6小鼠皮下注射PDAC细胞或尾静脉注射KPC肿瘤细胞。小鼠给予CCT 137690（80 mg/kg）或溶剂并监测肿瘤生长;收集肿瘤组织并通过免疫组织化学进行分析。我们使用在线R2基因组学分析和可视化平台比较了人类胰腺癌组织（n=130）与患者生存时间之间的基因表达水平。CCT 137690在PDAC细胞系中诱导坏死样死亡并减少集落形成;这些效应需要RIPK 1、RIPK 3和MLKL以及极光激酶A（AURKA）的抑制。AURKA直接与RIPK 1和RIPK 3相互作用以减少坏死体激活。AURKA介导的糖原合成酶激酶3 β（GSK 3B）在丝氨酸9处的磷酸化抑制了RIPK 3和MLKL坏死体的活化。AURKA（D274 A）或GSK 3 β（S9 A）突变，或通过RIPK 3和MLKL的RIPK 1信号传导的药理学抑制剂，降低了CCT 137690在PDAC细胞中的细胞毒性活性。经口给予CCT 137690可诱导小鼠皮下和原位肿瘤中的坏死性凋亡和免疫原性细胞死亡，并降低肿瘤生长和AURKA和GSK 3 β的肿瘤细胞磷酸化。CCT 137690增加了原位KPC PDAC小鼠的存活时间，并减少了肿瘤生长、基质和转移。胰腺癌患者AURKA和GSK 3 β mRNA表达增加与生存期缩短相关我们鉴定了极光激酶抑制剂CCT 137690作为一种通过RIPK 1、RIPK 3和MLKL诱导PDAC细胞坏死样死亡的药物。CCT 137690减缓了小鼠PDAC细胞原位肿瘤的生长，AURKA和GSK 3 β的表达与患者的生存时间相关。AURKA可能是治疗胰腺癌的靶点。
Induction of non-apoptotic cell death could be an approach to eliminate apoptosis-resistant tumors. We investigated necroptosis-based therapies in mouse models of pancreatic ductal adenocarcinoma cancer (PDAC). We screened 273 commercially available kinase inhibitors for cytotoxicity against a human PDAC cell line (PANC1). We evaluated the ability of the aurora kinase inhibitor CCT137690 to stimulate necroptosis in PDAC cell lines (PANC1, PANC2.03, CFPAC1, MiaPaCa2, BxPc3, and PANC02) and the HEK293 cell line, measuring loss of plasma membrane integrity, gain in cell volume, swollen organelles, and cytoplasmic vacuoles. We tested the effects of CCT137690 in colon formation assays, and the effects of the necroptosis (necrostatin-1 and necrosulfonamide), apoptosis, autophagy, and ferroptosis inhibitors. We derived cells from tumors that developed in Pdx1-Cre;K-RasG12D/+;p53R172H/+ (KPC) mice. Genes encoding proteins in cell death pathways were knocked out, knocked down, or expressed from transgenes in PDAC cell lines. Athymic nude or B6 mice were given subcutaneous injections of PDAC cells or tail-vein injections of KPC tumor cells. Mice were given CCT137690 (80 mg/kg) or vehicle and tumor growth was monitored; tumor tissues were collected and analyzed by immunohistochemistry. We compared gene expression levels between human pancreatic cancer tissues (n=130) with patient survival times using the online R2 genomics analysis and visualization platform. CCT137690 induced necrosis-like death in PDAC cell lines and reduced colony formation; these effects required RIPK1, RIPK3, and MLKL, as well as inhibition of aurora kinase A (AURKA). AURKA interacted directly with RIPK1 and RIPK3 to reduce necrosome activation. AURKA-mediated phosphorylation of glycogen synthase kinase 3 beta (GSK3B) at serine 9 inhibited activation of the RIPK3 and MLKL necrosome. Mutations in AURKA (D274A) or GSK3β (S9A), or pharmacologic inhibitors of RIPK1 signaling via RIPK3 and MLKL, reduced the cytotoxic activity of CCT137690 in PDAC cells. Oral administration of CCT137690 induced necroptosis and immunogenic cell death in subcutaneous and orthotopic tumors in mice, and reduced tumor growth and tumor cell phosphorylation of AURKA and GSK3β. CCT137690 increased survival times of mice with orthotopic KPC PDACs and reduced tumor growth, stroma, and metastasis. Increased expression of AURKA and GSK3β mRNAs associated with shorter survival times of patients with pancreatic cancer. We identified the aurora kinase inhibitor CCT137690 as an agent that induces necrosis-like death in PDAC cells, via RIPK1, RIPK3, and MLKL. CCT137690 slowed growth of orthotopic tumors from PDAC cells in mice, and expression of AURKA and GSK3β associate with patient survival times. AURKA might be targeted for treatment of pancreatic cancer.
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影响因子：
50.3
作者：
Katayama H;Wang J;Treekitkarnmongkol W;Kawai H;Sasai K;Zhang H;Wang H;Adams HP;Jiang S;Chakraborty SN;Suzuki F;Arlinghaus RB;Liu J;Mobley JA;Grizzle WE;Wang H;Sen S
通讯作者：
Sen S
影响因子：
8.8
作者：
Dondelinger, Yves;Declercq, Wim;Vandenabeele, Peter
通讯作者：
Vandenabeele, Peter
影响因子：
7.3
作者：
Bavetsias, Vassilios;Large, Jonathan M.;McDonald, Edward
通讯作者：
McDonald, Edward
慢性淋巴细胞白血病中 TNFα 诱导的坏死性凋亡信号失调：LEF1 抑制 CYLD 基因
影响因子：
11.4
作者：
Liu, P.;Xu, B.;Li, J.
通讯作者：
Li, J.
影响因子：
--
作者：
Jacobs KM;Bhave SR;Ferraro DJ;Jaboin JJ;Hallahan DE;Thotala D
通讯作者：
Thotala D