单细胞RNA测序揭示血管内皮生长因子信号传导介导非小细胞肺癌中对新辅助安罗替尼联合程序性死亡受体1阻断疗法的反应。
scRNA-seq reveals that VEGF signaling mediates the response to neoadjuvant anlotinib combined with PD-1 blockade therapy in non-small cell lung cancer.
作者信息
Huang Ziqi, Li Li, Zhao Xiaohe, Jin Haixia, Shen Meng, Li Baihui, Zeng Yu, Zhang Qinfen, Wang Qiyu, Wang Meng, Yang Lili
机构信息
Department of Immunology, Tianjin's Clinical Research Center for Cancer, Key Laboratory of Cancer Immunology and Biotherapy, State Key Laboratory of Druggability Evaluation and Systematic Translational Medicine, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Huanhuxi Road, Tiyuanbei, Hexi District, Tianjin, 300060, China.
Department of Esophageal Cancer, Tianjin's Clinical Research Center for Cancer, Key Laboratory of Cancer Prevention and Therapy, Tianjin Medical University Cancer Institute and Hospital, National Clinical Research Center for Cancer, Tianjin, China.
出版信息
J Transl Med. 2025 Apr 25;23(1):478. doi: 10.1186/s12967-025-06485-4.
BACKGROUND
Clinical trials have shown that neoadjuvant anlotinib combined with PD-1 blockade therapy can prolong the survival of patients with driver gene negative non-small cell lung cancer (NSCLC), but some patients fail to benefit from the combination therapy.
METHODS
To explore the potential drug resistance mechanism and predict the efficacy of neoadjuvant therapy in NSCLC patients, we used scRNA-seq to observe and analyze the dynamic changes of immune cells, stromal cells and cancer cells in NSCLC patients who received neoadjuvant combination therapy. We analyzed transcriptome data of ~ 47,000 single cells from 9 NSCLC patients, including 3 treatment naïve patients, 3 post-treatment patients with major pathological response (MPR), and 3 Non-MPR patients. Subsequently, the infiltration of immune cells was detected by immunohistochemistry and multiplex immunofluorescence in NSCLC.
RESULTS
In MPR patients, we found that neoadjuvant therapy reduced the expression of the T cell exhausted signature, reduced the transition of T_THEMIS cells to Tregs, and enhanced the positive feedback between CD4 T cells and PAX5 memory B cells. In Non-MPR patients, tumor-associated macrophages (TAMs) dampen therapeutic efficiency by being the hub of cell communication. TAMs and fibroblasts stimulate endothelial cells via VEGF, endothelial ZEB1 may up-regulate FLT1 (VEGFR) expression in response to anlotinib, and VEGFR endothelial cell signature can predict survival of NSCLC cohort in TCGA. In addition, PLA2G4A, the key enzyme in the VEGF pathway, was highly expressed in the tumor cells of Non-MPR patients after anlotinib treatment. In 135 NSCLC patients, we confirmed by immunohistochemistry that PLA2G4A was positively correlated with poor prognosis and Tregs infiltration.
CONCLUSION
In conclusion, VEGF signaling dependent dynamic changes in endothelial and epithelial cells are deeply involved in the formation of anlotinib resistance and immunosuppression phenotypes in NSCLC patients.
背景
临床试验表明，新辅助安罗替尼联合程序性死亡受体1（PD-1）阻断疗法可延长驱动基因阴性的非小细胞肺癌（NSCLC）患者的生存期，但部分患者未能从联合治疗中获益。
方法
为探究NSCLC患者新辅助治疗的潜在耐药机制并预测其疗效，我们采用单细胞RNA测序（scRNA-seq）观察并分析接受新辅助联合治疗的NSCLC患者免疫细胞、基质细胞和癌细胞的动态变化。我们分析了9例NSCLC患者约47,000个单细胞的转录组数据，其中包括3例未经治疗的患者、3例出现主要病理缓解（MPR）的治疗后患者和3例未出现MPR的患者。随后，通过免疫组化和多重免疫荧光检测NSCLC中免疫细胞的浸润情况。
结果
在出现MPR的患者中，我们发现新辅助治疗降低了T细胞耗竭标志物的表达，减少了T_THEMIS细胞向调节性T细胞（Tregs）的转变，并增强了CD4 T细胞与PAX5记忆B细胞之间的正反馈。在未出现MPR的患者中，肿瘤相关巨噬细胞（TAM）作为细胞通讯的枢纽降低了治疗效果。TAM和成纤维细胞通过血管内皮生长因子（VEGF）刺激内皮细胞，内皮细胞中的锌指蛋白E盒结合因子1（ZEB1）可能会响应安罗替尼而上调Fms样酪氨酸激酶1（FLT1，即VEGFR）的表达，并且VEGFR内皮细胞标志物可以预测癌症基因组图谱（TCGA）中NSCLC队列的生存期。此外，VEGF通路中的关键酶磷脂酶A2G4A（PLA2G4A）在接受安罗替尼治疗后的未出现MPR患者的肿瘤细胞中高表达。在135例NSCLC患者中，我们通过免疫组化证实PLA2G4A与预后不良和Tregs浸润呈正相关。
结论
总之，内皮细胞和上皮细胞中依赖VEGF信号的动态变化深度参与了NSCLC患者安罗替尼耐药和免疫抑制表型的形成。
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