PO.CL01.22 · 临床研究
转移性胰腺癌中Kras活性关联的糖酵解与免疫抑制
Kras activity-linked glycolysis and immunosuppression in metastatic pancreatic cancer
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：KRAS突变作为胰腺导管腺癌（PDAC）的驱动因素，涉及ERK依赖性基因转录。抑制突变型KRAS活性可抑制糖酵解并重塑小鼠PDAC中不良的免疫谱。在人类PDAC中识别这些关系有助于揭示代谢和免疫依赖性的关键驱动因素。近期鉴定的PDAC siKRAS、KRASi、iKras基因集使得能够创建反映KRAS活性的精确KRAS特征评分。利用肝转移（LM）来源的转录组数据，我们评估了KRAS特征评分对临床结局及转移性PDAC分子特征的影响。
方法：使用来自未经治疗的PDAC肝转移灶的针刺活检样本，在cDNA微阵列上测量了全面的mRNA表达，并通过反相蛋白质阵列（RPPA）测量了435种蛋白表达。计算每个样本的KRAS特征评分。基于KRAS特征评分与加权共表达网络分析所得模块特征基因之间的关系确定KRAS相关基因模块。对RPPA数据进行层次聚类生成KRAS相关亚型，其中从KRAS相关基因模块的基因所编码的分子中选出KRAS相关蛋白。
结果：在77例患者（男性：65%；中位年龄：66岁）中计算了LM的KRAS特征评分。KRAS特征评分高的肿瘤患者（n=38）与评分低的患者（n=39）相比总生存较差（风险比：1.895 [95%置信区间：1.123至3.198]）。KRAS突变的LM（90.9%）与高KRAS特征评分相关（P=0.004）。DUSP4 mRNA表达（一种用于ERK去磷酸化的ERK依赖性负反馈基因）在高KRAS特征评分的肿瘤（P=0.017）及KRAS突变的肿瘤（P<0.001）中升高。37种蛋白被鉴定为KRAS相关上调蛋白，包括参与糖酵解通路的蛋白及乳酸转运体MCT4。在HK2（P=0.132）、Eno1、2（P<0.001，P=0.016）、PKM（P<0.001）、LDHA（P<0.001）和MCT4（P<0.001）中发现糖酵解和乳酸外排通路的基因上调。ZAP-70（一种T细胞受体信号传导所必需的胞质酪氨酸激酶）位列九种KRAS相关下调蛋白之中。在CD4（P<0.001）、CD8A（P<0.001）、CD8B（P=0.013）和ZAP-70（P<0.001）中发现免疫谱的基因下调。
结论：KRAS上调的肿瘤在胰腺癌肝转移中表现出糖酵解分子升高和免疫抑制。KRAS活性可能驱动代谢和免疫的恶化，导致不良预后。
查看英文原文 English abstract
Background: KRAS mutations act as a driver of pancreatic ductal adenocarcinoma (PDAC), involving ERK-dependent gene transcription. Inhibition of mutated KRAS activity suppresses glycolysis and reshapes the poor immune profile in murine PDAC. Identification of these relationships in human PDAC is useful for realizing pivotal drivers of metabolic and immune dependency. Recently identified PDAC siKRAS KRASi iKras gene sets enable the creation of a precise KRAS signature score for KRAS activity. Using liver metastasis (LM)-derived transcriptome data, we evaluated the effect of KRAS signature score on clinical outcomes and the molecular features of metastatic PDAC.
Methods: Comprehensive mRNA expression on cDNA microarray and 435 protein expressions on reverse-phased protein array (RPPA) were measured using needle-biopsied samples from treatment-naïve LM of PDAC. KRAS signature score was calculated in each sample. A KRAS-related gene module was determined based on the relationship between KRAS signature score and module eigengene from weighted co-expression network analysis. Hierarchical clustering of RPPA data generated KRAS-related subtypes, in which a KRAS-related protein was selected from the molecule encoded by the genes of KRAS-related gene modules.
Results: KRAS signature score in LMs was calculated in 77 patients (Male: 65%; median age: 66 years). Patients with high KRAS signature score tumors (n=38) showed poor overall survival as compared to those with low KRAS signature score tumors (n=39; hazard ratio: 1.895 [95% confidential interval: 1.123 to 3.198]). KRAS mutated LMs (90.9%) were related to high KRAS signature scores (P=0.004). DUSP4 mRNA expression, an ERK-dependent negative feedback gene for ERK dephosphorylation, increased in tumors with high KRAS signature scores (P=0.017) and with KRAS mutation (P<0.001). Thirty-seven proteins were identified as KRAS-related UP proteins, including those involved in the glycolysis pathway and a lactate transporter, MCT4. The gene up-regulation of glycolysis and lactate-efflux pathway was found in HK2 (P=0.132), Eno1, 2 (P<0.001, P=0.016), PKM (P<0.001), LDHA (P<0.001), and MCT4 (P<0.001). ZAP-70, a cytosolic tyrosine kinase essential for T cell receptor signaling, was present in nine KRAS-related DOWN proteins. The gene down-regulation of the immune profile was found in CD4 (P<0.001), CD8A (P<0.001), CD8B (P=0.013), and ZAP-70 (P<0.001).
Conclusion: KRAS up-regulated tumors showed elevation of glycolytic molecules and immunosuppression in LM of pancreatic cancer. KRAS activity might drive metabolic and immunological aggravation, leading to a poor prognosis.
利益披露 Disclosure
S. Mitsunaga,
Toray Industries, Inc ).
Ajinomoto Co., Inc ).
Mitsui Chemicals, Inc ).
Taiho Innovations, LLC ).
Taiho Pharmaceutical Co., Ltd ).
Tensegrity Pharma, Inc ).
Pfizer Japan Inc ).
Ono Pharmaceutical Co., Ltd. ).
Astellas Pharma Inc ).
Chugai Pharmaceutical Co., Ltd. ), Patent.
K. Kato, None..
H. Suzuki, None.
M. Sasaki,
AstraZeneca ).
Boehringer Ingelheim ).
MSD ).
Nihon Servier ).
Taiho ).
H. Yoshimatsu,
Pfizer R&D Japan G.K. Employment.
Y. Kamei,
Pfizer R&D Japan G.K. Employment.
M. Ikeda,
AstraZeneca ).
Abbvie ).
Amgen ).
Bristol Myers Squibb ).
Chugai ).
Chiome Bioscience ).
Delta-Fly Pharma ).
Eisai ).
Eli Lilly Japan ).
Invitae ).
MSD ).
J-Pharma ).
Merck biopharma ).
Merus N.V. ).
Novartis ).
Nihon Servier ).
Ono ).
Syneos Health ).
Rakuten Medical ).
Taiho ).