Malignant cell-specific CXCL14 promotes tumor lymphocyte infiltration in oral cavity squamous cell carcinoma.
Malignant cell-specific CXCL14 promotes tumor lymphocyte infiltration in oral cavity squamous cell carcinoma.
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DOI：
10.1136/jitc-2020-001048
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发表时间：
2020-09
影响因子：
10.9
通讯作者：
Puram SV
中科院分区：
文献类型：
--
作者：
Parikh A;Shin J;Faquin W;Lin DT;Tirosh I;Sunwoo JB;Puram SV
探索淋巴细胞浸润作为 CXCL14 介导的口腔鳞状细胞癌 (OSCC) 肿瘤生长抑制背后的潜在机制。我们分析了来自 OSCC 的单细胞 RNA 测序 (scRNA-seq) 数据，以确定淋巴结 (LN) 恶性细胞与原发肿瘤之间的表达变化。使用 qRT-PCR 定量小鼠 OSCC 细胞系中的 CXCL14 表达。在小鼠口腔（MOC）1细胞中进行CXCL14的短发夹RNA敲除，并在MOC2细胞中进行CXCL14过表达。将每种条件下的细胞注射到去除和未去除 T 细胞的 C57BL/6 小鼠中，并测量肿瘤体积。 30天时，分离肿瘤并通过流式细胞术分析CD45+CD3+T细胞。 CXCL14 表达与 scRNA-seq 数据中肿瘤浸润淋巴细胞 (TIL) 以及 TCGA 肿瘤的基因表达特征相关。 scRNA-seq 显示 CXCL14 是 LN 中恶性细胞中相对于原发肿瘤最显着下调的基因，支持其在预防侵袭和/或转移中的作用。在小鼠免疫活性模型中，惰性 MOC1 细胞中的 CXCL14 表达高于更具攻击性的 MOC2 细胞。与对照相比，MOC1 细胞中 CXCL14 敲低显着增加了体内肿瘤生长，TIL 相应减少。在 MOC2 细胞中，相对于对照，CXCL14 过表达显着降低了肿瘤生长，并且 TIL 增加。这两种效应都会随着 T 细胞的耗竭而消失。在人类肿瘤 scRNA-seq 队列中，我们发现只有恶性细胞 CXCL14 与 TIL 相关，而非恶性细胞或成纤维细胞 CXCL14 与 TIL 相关。 TCGA 队列中的大量 CXCL14 与 TIL 没有关联。肿瘤细胞较高的 CXCL14 表达与肿瘤生长减少和 TIL 增加相关，支持免疫介导的 OSCC 肿瘤生长抑制。鉴于与原发肿瘤相比，CXCL14 在淋巴结转移中下调，我们的数据提出了 CXCL14 介导的免疫浸润可能阻止侵袭和转移的可能性。在人类 scRNA-seq 数据中，只有恶性细胞特异性 CXCL14 与 TIL 相关，这表明 CXCL14 表达具有关键的背景依赖性效应。
To explore lymphocyte infiltration as a potential mechanism behind CXCL14-mediated tumor growth suppression in oral cavity squamous cell carcinoma (OSCC). We analyzed single cell RNA-sequencing (scRNA-seq) data from OSCC to identify expression changes among malignant cells in lymph nodes (LN) versus primary tumors. CXCL14 expression in murine OSCC cell lines was quantified using qRT-PCR. Short hairpin RNA knockdown of CXCL14 was performed in mouse oral cavity (MOC)1 cells, and CXCL14 overexpression was performed in MOC2 cells. Cells in each condition were injected into C57BL/6 mice with and without T cell depletion, and tumor volume was measured. At 30 days, tumors were dissociated and analyzed by flow cytometry for CD45+CD3+ T cells. CXCL14 expression was correlated with gene expression signatures of tumor infiltrating lymphocytes (TIL) in scRNA-seq data, as well as TCGA tumors. scRNA-seq revealed CXCL14 as the most significantly downregulated gene among malignant cells in LNs relative to primary tumor, supporting a role in preventing invasion and/or metastasis. In a murine immunocompetent model, CXCL14 expression was higher in indolent MOC1 cells than in more aggressive MOC2 cells. Tumor growth in vivo was significantly increased by CXCL14 knockdown in MOC1 cells relative to control, with a corresponding decrease in TIL. In MOC2 cells, tumor growth was significantly reduced by CXCL14 overexpression relative to control and TIL were increased. Both effects were lost with T cell depletion. In a human tumor scRNA-seq cohort, we found that only malignant cell CXCL14, but not non-malignant cell or fibroblast CXCL14, was associated with TIL. Bulk CXCL14 from the TCGA cohort had no association with TIL. Higher CXCL14 expression by tumor cells is associated with reduced tumor growth and increased TIL, supporting immune-mediated suppression of tumor growth in OSCC. Given that CXCL14 is downregulated in LN metastases compared with primary tumors, our data raise the possibility that CXCL14-mediated immune infiltration may discourage invasion and metastasis. In human scRNA-seq data, only malignant cell-specific CXCL14 was associated with TIL, suggesting a critical context-dependent effect of CXCL14 expression.