PO.TB10.04 · 肿瘤生物学
评估放化疗对HNSCC肿瘤微环境衰老状态的影响
Evaluation of chemo-radiation therapy's effect on the senescence status of tumor microenvironment of HNSCC
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
背景：细胞衰老是一种应激诱导的、不可逆的细胞增殖停滞，伴随衰老相关分泌表型（SASP），这可能损害免疫系统检测和清除癌细胞的能力。化疗和放疗（RT）通过诱导DNA损伤，是头颈部鳞状细胞癌（HNSCC）的标准治疗。尽管具有治疗价值，但研究表明，化疗和RT诱导的DNA损伤在其他癌症中也可能导致免疫细胞衰老。这些疗法对HNSCC肿瘤微环境（TME）衰老状态的影响尚未阐明。我们假设放化疗会导致HNSCC中TME的免疫衰老。在本研究中，我们分析了化疗和放疗如何影响HNSCC的TME中衰老相关基因程序。
方法：从30例HNSCC患者放化疗前后获得了批量免疫靶向RNA测序数据（来自GEO的GSE193445）。从4例HNSCC口腔癌肿瘤样本（GSE280982）在RT治疗的3个时间点获得了单细胞RNA测序数据。对测序数据进行了过滤、标准化、整合和聚类。使用生成式AI辅助调试分析。
结果：批量RNA-seq数据的基因集富集分析（GSEA）显示，放化疗后，参与免疫应答的髓系细胞活化上调（NES = +1.93，padj = 0.0056），而胞质分裂下调（NES = -2.32，padj = 0.0037）。差异表达分析鉴定出干扰素刺激基因ISG15（log2FC = -1.48，padj = 4.9e-07）和IFI6（log2FC = -1.46，padj = 2.2e-07）在放化疗后强烈下调。其他免疫调节基因表达如IGFBP3（log2FC = -1.56，padj = 6.0e-08）和GAGE家族基因（log2FC = -1.79，padj = 0.022）在治疗后也降低。单细胞RNA-Seq数据显示，RT后在各个细胞簇中，在CD8+ T细胞中，p53通路上调（NES = 1.8，padj = 0.0038），IL-2/STAT5信号通路受到抑制（NES = -1.8，padj = 0.02）。在B细胞中，TNFalpha信号（NES = 2.6，padj = 4.1e-07）和凋亡通路（NES = 1.66，padj = 0.03）上调。在Treg中，氧化磷酸化（NES = 2.6，padj = 6.1e-05）和MYC信号在Treg中上调（NES = 3.2，padj = 2.5e-08）。
结论：初步数据表明，放化疗后HNSCC的TME中先天免疫通路受到抑制；CD8+ T细胞、B细胞和Treg的增殖和存活通路作为对RT的响应而发生改变。
查看英文原文 English abstract
Background: Cellular senescence is a stress-induced, irreversible arrest of cell proliferation accompanied by a senescence-associated secretory phenotype (SASP), which may impair the immune system's ability to detect and eliminate cancer cells. Chemotherapy and radiation therapy (RT) are standard treatments for head and neck squamous cell carcinoma (HNSCC) by inducing DNA damage. Despite their therapeutic utility, studies have shown that chemotherapy and RT-induced DNA damage may also cause immune cell senescence in other cancers. The effect of these therapies on the senescence state of the tumor microenvironment (TME) in HNSCC has not been elucidated. We hypothesized that chemoradiation treatment results in the immunosenescence of TME in HNSCC. In this study, we analyzed how chemotherapy and radiation therapy influence the senescence-associated gene programs in the TME of HNSCC.
Methods: Bulk immune-targeted RNA sequencing data were obtained from 30 HNSCC patients before and after chemoradiotherapy (GSE193445 from GEO). Single-cell RNA sequencing data were obtained from oral cancer tumor samples of 4 patients with HNSCC (GSE280982) at 3 time points of RT treatment. Filtering, normalization, integration, and clustering were performed on the sequencing data. Generative AI was used for assistance in debugging analyses.
Results: Gene Set Enrichment Analysis (GSEA) of bulk RNA-seq data shows that, post-chemoradiation, myeloid cell activation involved in immune response was upregulated (NES = +1.93, padj = 0.0056), while cytokinesis was downregulated (NES = -2.32, padj = 0.0037). Differential expression analysis identified interferon-stimulated genes ISG15 (log2FC = -1.48, padj = 4.9e-07) and IFI6 (log2FC = -1.46, padj = 2.2e-07) were strongly downregulated after chemoradiation. Additional immune-modulatory genes expression such as IGFBP3 (log2FC = -1.56, padj = 6.0e-08) and GAGE family genes (log2FC = -1.79, padj = 0.022) also decreased post-treatment. Single-cell RNA-Seq data show that in individual cell clusters after RT, in CD8+ T cells, the p53 pathway was upregulated (NES = 1.8, padj = 0.0038) and IL-2/STAT5 signaling pathway was suppressed (NES = -1.8, padj = 0.02). In B cells, TNFalpha signaling (NES = 2.6, padj = 4.1e-07) and apoptosis pathway (NES = 1.66, padj = 0.03) is upregulated. In Tregs, oxidative phosphorylation (NES = 2.6, padj = 6.1e-05) and MYC signaling is upregulated in Tregs (NES = 3.2, padj = 2.5e-08).
Conclusion: Preliminary data suggest that innate immune pathways in the TME of HNSCC were suppressed after chemoradiation therapy; the proliferative and survival pathways of CD8+ T cells, B cells, and Tregs are altered as a response to RT.
利益披露 Disclosure
L. Gao, None..
A. Sun, None..
R. Xin, None..
D. John, None..
W. Li, None..
W. M. Ongkeko, None.