Intra-tumor immune infiltration is a crucial element interacting with tumor cells in intrahepatic cholangiocarcinoma (ICC). However, its phenotype and related spatial structure remained elusive. To address these limitations, we undertook a comprehensive study combining spatial data (29,632 spots from six samples) and single-cell data (21,158 cells from 35 samples). We identified two distinct infiltration patterns: macrophage+ (characterized by CD68 and MARCO) and plasma cell+ (characterized by IGHG1 and JCHAIN). The macrophage+ and plasma cell+ signatures showed adverse and favorable roles in ICC patients’ survival, respectively. Notably, MARCO+ tumor-associated macrophage (TAM) was recognized as the main cell type in macrophage+ samples, indicating an immune-resistant microenvironment. Increased epithelial-mesenchymal transition activities, angiogenesis, and hypoxia were observed in MARCO+ TAM. The co-location of MARCO+ TAM and CTSE+ tumor cells was observed in spatial transcriptomics and bulk transcriptomics data, validated by multiplex immunofluorescence performed on twenty ICC samples. The co-location area exhibited similar protumorigenic pathways and suppressed immune response. CTSE exhibited associations with intrahepatic metastasis and vascular invasion. Both MARCO+ TAM and CTSE+ tumor cells were associated with worse survival and patients with high infiltration of two cell types displayed the worst survival. Within the co-location area, the galectin signaling pathway was most active in cell-cell communication, with LGALS9-CD44 identified as the main ligand-receptor pair. This study identified macrophage+ and plasma cell+ intra-tumor immune infiltration patterns and the co-location of MARCO+ TAM and CTSE+ tumor cells as contributors to immune resistance.

中文翻译：

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MARCO + 肿瘤相关巨噬细胞和 CTSE+ 肿瘤细胞的共定位决定了肝内胆管癌的不良预后


肿瘤内免疫浸润是肝内胆管癌 （ICC） 中与肿瘤细胞相互作用的关键因素。然而，其表型和相关空间结构仍然难以捉摸。为了解决这些限制，我们进行了一项综合研究，结合了空间数据（来自 6 个样本的 29,632 个点）和单细胞数据（来自 35 个样本的 21,158 个细胞）。我们确定了两种不同的浸润模式：巨噬细胞+ （以 CD68 和 MARCO 为特征） 和浆细胞 + （以 IGHG1 和 JCHAIN 为特征）。巨噬细胞 + 和浆细胞 + 特征分别在 ICC 患者生存中显示出不良和有利作用。值得注意的是，MARCO + 肿瘤相关巨噬细胞 （TAM） 被认为是巨噬细胞 + 样品中的主要细胞类型，表明免疫耐药微环境。在 MARCO + TAM 中观察到上皮-间充质转化活性增加、血管生成和缺氧。在空间转录组学和大量转录组学数据中观察到 MARCO + TAM 和 CTSE + 肿瘤细胞的共定位，并通过对 20 个 ICC 样本进行的多重免疫荧光验证。共定位区域表现出相似的促瘤途径并抑制免疫反应。CTSE 与肝内转移和血管浸润有关。MARCO+ TAM 和 CTSE+ 肿瘤细胞均与较差的生存率相关，两种细胞类型高浸润的患者生存率最差。在共定位区域内，半乳糖凝集素信号通路在细胞间通讯中最活跃，LGALS9-CD44 被确定为主要配体-受体对。本研究确定了巨噬细胞 + 和浆细胞 + 肿瘤内免疫浸润模式以及 MARCO+ TAM 和 CTSE+ 肿瘤细胞的共定位是免疫耐药的促成因素。