SPINK1-driven cellular heterogeneity and interaction networks in intrahepatic cholangiocarcinoma revealed by integrated multi-omics analysis

作者：Xiaoxiang Rong, Jiayun Guan, Chanqi Ye, Qiong Li, Mingyu Wan, Wenguang Fu, Jinzhang Chen, Dayong Zheng, Ruyin Chen, Rui Zhou, Gautam Sethi, Yunlu Jia, Jian Ruan · 发表于：Journal of Advanced Research · 年份：2025 · DOI：10.1016/j.jare.2025.11.011 · 被引用次数：1 · 研究领域：Cholangiocarcinoma and Gallbladder Cancer Studies、Chemokine receptors and signaling、Ferroptosis and cancer prognosis

INTRODUCTION: Intrahepatic cholangiocarcinoma (ICC) is a highly heterogeneous and aggressive malignancy with poor prognosis and limited treatment options. The lack of precise molecular classification and incomplete understanding of the tumor microenvironment (TME) impede therapeutic development. OBJECTIVES: This study aims to dissect the cellular heterogeneity and intercellular interactions in ICC, with a particular focus on SPINK1-overexpressing epithelial subsets. We sought to determine the role of SPINK1 in modulating immune cell recruitment and tumor metabolism, and to evaluate its clinical relevance. METHODS: We performed single-cell RNA sequencing (scRNA-seq) on human ICC tumors and adjacent normal tissues to construct a transcriptomic atlas. Integrative proteomic analysis, transwell assays, co-culture systems, and functional perturbation experiments (SPINK1 knockdown and ASCT2 inhibition) were conducted to explore epithelial-macrophage interactions and metabolic dynamics. RESULTS: ScRNA-seq identified a SPINK1-overexpressing epithelial subcluster enriched in ICC tumors. High SPINK1 expression correlated with significantly shorter patient survival. SPINK1-overexpressing epithelial subcluster secretes CCL20, which recruits lipid-associated macrophages (LAMs). This effect was reversed by CCL20 neutralizing antibodies. In co-culture, LAMs increased intracellular glutamine levels in SPINK1-overexpressing epithelial subcluster, promoting proliferation. Disruption of SPINK1 e...