PO.ET03.02 · 实验与分子治疗
幽门螺杆菌感染诱导WEE1促进胃腺癌的耐药
H. pylori infection induces WEE1 to promote drug resistance in gastric adenocarcinoma
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：胃癌（GC）是全球癌症相关死亡的第5大原因，预后不良且经常出现治疗耐药。迫切需要鉴定这种耐药的关键分子驱动因素和新的治疗易感靶点。本研究探讨核激酶WEE1在GC进展和耐药中的非经典作用，尤其是在幽门螺杆菌（Helicobacter pylori, H. pylori）感染背景下。
方法：采用对来自各种数据集、细胞系模型和患者来源异种移植（PDX）的2,000余份胃癌患者样本的整合分析，以阐明WEE1相关的信号通路。进行基因集富集分析（GSEA）以鉴定与高WEE1表达相关的转录程序。使用siRNA敲低、过表达、Western blot、RT-qPCR、免疫荧光和体内异种移植模型进行功能验证。对892种FDA批准的药物进行高通量筛选，以鉴定与WEE1抑制剂MK1775有效的药物组合。
结果：发现WEE1在胃癌细胞中异常过表达并错误定位于细胞质。跨14个数据集的GSEA一致显示WEE1高表达肿瘤中MYC和E2F靶基因的富集。机制上，WEE1通过磷酸化RB激活E2F1，导致MYC的转录上调。MYC反过来增强WEE1和E2F1的表达，形成一个前馈致癌环路。值得注意的是，H. pylori感染强力诱导WEE1和MYC表达及其下游效应因子，将微生物病因与转录重编程和肿瘤进展联系起来。用MK1775靶向WEE1抑制了MYC驱动的转录活性，降低了抗凋亡基因表达（BCL2、BCL2L1），并使GC细胞对DNA损伤剂敏感。高通量筛选鉴定出Afatinib（一种EGFR/HER2抑制剂）为MK1775的强效协同伙伴。联合治疗在体外、PDX模型以及源自GC患者的3D类器官系统中均表现出增强的抗肿瘤疗效。
结论：本研究鉴定出一条新的H. pylori-WEE1-E2F1-MYC信号轴，促成GC的发病机制和耐药。鉴于缺乏临床可用的MYC抑制剂，靶向WEE1提供了一种有前景的替代方法。与Afatinib协同疗效的发现支持了一种克服MYC驱动胃癌耐药的新治疗策略。
查看英文原文 English abstract
Background: Gastric cancer (GC) is the 5 th leading cause of cancer-related mortality worldwide, with a poor prognosis and frequent resistance to therapy. Identifying key molecular drivers of this resistance and novel therapeutic vulnerabilities is urgently needed. This study investigates the non-canonical role of the nuclear kinase WEE1 in GC progression and drug resistance, particularly in the context of Helicobacter pylori (H. pylori) infection.
Methods: Integrative analyses of over 2,000 gastric cancer patient samples from various datasets, cell line models, and patient-derived xenografts (PDXs) were employed to elucidate WEE1-associated signaling pathways. Gene set enrichment analysis (GSEA) was conducted to identify transcriptional programs associated with high WEE1 expression. Functional validation was performed using siRNA knockdown, overexpression, Western blot, RT-qPCR, immunofluorescence, and in vivo xenograft models. A high-throughput screen of 892 FDA-approved drugs was conducted to identify effective drug combinations with the WEE1 inhibitor MK1775.
Results: WEE1 was found to be aberrantly overexpressed and mis-localized to the cytoplasm in gastric cancer cells. GSEA across 14 datasets consistently showed enrichment of MYC and E2F target genes in WEE1-high tumors. Mechanistically, WEE1 activated E2F1 through the phosphorylation of RB, leading to the transcriptional upregulation of MYC. MYC, in turn, enhanced expression of WEE1 and E2F1, forming a feedforward oncogenic loop. Notably, H. pylori infection robustly induced WEE1 and MYC expression, as well as their downstream effectors, linking microbial etiology to transcriptional reprogramming and tumor progression. Targeting WEE1 with MK1775 suppressed MYC-driven transcriptional activity, reduced anti-apoptotic gene expression (BCL2, BCL2L1), and sensitized GC cells to DNA-damaging agents. A high-throughput screen identified Afatinib, an EGFR/HER2 inhibitor, as a potent synergistic partner with MK1775. Combination therapy demonstrated enhanced anti-tumor efficacy in vitro , in PDX models, and in 3D organoid systems derived from GC patients.
Conclusions: This study identifies a novel H. pylori -WEE1-E2F1-MYC signaling axis that contributes to GC pathogenesis and resistance. Given the lack of clinically available MYC inhibitors, targeting WEE1 provides a promising alternative approach. The discovery of synergistic efficacy with Afatinib supports a new therapeutic strategy for overcoming resistance in MYC-driven gastric cancer.
利益披露 Disclosure
M. Islam, None.