肝胆管癌 (H-ChC) 具有肝细胞癌 (HCC) 和肝内胆管癌 (iCCA) 的临床病理学特征，是比 HCC 或 iCCA 更具侵袭性的原发性肝癌亚型。我们对 16 个人类样本中的 91,112 个单细胞转录组进行了测序，阐明H-ChC中HCC和iCCA成分共存的分子机制。我们在全转录组水平上观察到H-ChC的两种分子亚型（CHP和CIP），其中富含CHP的代谢活跃的肿瘤细胞亚群被表征通过细胞预分化特性。为了定义肿瘤及其相关微环境的异质性，我们观察到 H-ChC 中的肿瘤多样性比 HCC 和 iCCA 更大。与 HCC 和 iCCA 相比，H-ChC 表现出较弱的免疫细胞浸润和更大的 CD8 耗尽 T 细胞 (Tex) 功能障碍。然后，我们定义了 6,852 个 CD8 Tex  细胞的两种广泛细胞状态：GZMK CD8 Tex 细胞和末端 CD8 Tex 细胞。 GZMK CD8 Tex 细胞在 H-ChC 中治疗后表现出更高的浸润性，免疫治疗后其效应子评分和免疫检查点表达大大增加，这表明 H-ChC 可能比 HCC 或 iCCA 对免疫治疗更敏感。论文对H-ChC进行了探索，希望能够为其他癌症中混合瘤的研究做出贡献。2024肝胆外科与营养。版权所有。

Hepatocholangiocarcinoma (H-ChC) has the clinicopathological features of both hepatocellular carcinoma (HCC) and intrahepatic cholangiocarcinoma (iCCA) and is a more aggressive subtype of primary hepatic carcinoma than HCC or iCCA.We sequenced 91,112 single-cell transcriptomes from 16 human samples to elucidate the molecular mechanisms underlying the coexistence of HCC and iCCA components in H-ChC.We observed two molecular subtypes of H-ChC at the whole-transcriptome level (CHP and CIP), where a metabolically active tumour cell subpopulation enriched in CHP was characterized by a cellular pre-differentiation property. To define the heterogeneity of tumours and their associated microenvironments, we observe greater tumour diversity in H-ChC than HCC and iCCA. H-ChC exhibits weaker immune cell infiltration and greater CD8+ exhausted T cell (Tex) dysfunction than HCC and iCCA. Then we defined two broad cell states of 6,852 CD8+ Tex cells: GZMK+ CD8+ Tex cells and terminal CD8+ Tex cells. GZMK+ CD8+ Tex cells exhibited higher infiltration of after treatment in H-ChC, the effector scores and expression of the immune checkpoints of them greatly increased after immunotherapy, which indicated that H-ChC might be more sensitive than HCC or iCCA to immunotherapy.In this paper, H-ChC was explored, hoping to contribute to the study of mixed tumours in other cancers.2024 Hepatobiliary Surgery and Nutrition. All rights reserved.