Single-cell dissection of remodeled inflammatory ecosystem in primary and metastatic gallbladder carcinoma

Xiang Wang; Chunliang Liu; Jianan Chen; Lei Chen; Xianwen Ren; Minghui Hou; Xiuliang Cui; Youhai Jiang; Erdong Liu; Yali Zong; Anqi Duan; Xiaohui Fu; Wenlong Yu; Xiaofang Zhao; Zhao Yang; Yongjie Zhang; Jing Fu; Hongyang Wang

doi:10.1038/s41421-022-00445-8

Single-cell dissection of remodeled inflammatory ecosystem in primary and metastatic gallbladder carcinoma

Cell Discov. 2022 Oct 5;8(1):101. doi: 10.1038/s41421-022-00445-8.

Authors

Xiang Wang^#^{1

2}, Chunliang Liu^#¹, Jianan Chen^#¹, Lei Chen^#¹, Xianwen Ren³, Minghui Hou⁴, Xiuliang Cui¹, Youhai Jiang⁵, Erdong Liu⁶, Yali Zong⁶, Anqi Duan², Xiaohui Fu², Wenlong Yu², Xiaofang Zhao⁴, Zhao Yang⁷, Yongjie Zhang⁸, Jing Fu⁹, Hongyang Wang¹⁰

Affiliations

¹ International Cooperation Laboratory on Signal Transduction, National Center for Liver Cancer, Ministry of Education Key Laboratory on Signaling Regulation and Targeting Therapy of Liver Cancer, Shanghai Key Laboratory of Hepato-biliary Tumor Biology, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China.
² Second Department of Biliary Surgery, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China.
³ Changping Laboratory, Yard 28, Science Park Road, Changping District, Beijing, China.
⁴ Research Center for Organoids, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, Henan, China.
⁵ Cancer Research Center, The First Affiliated Hospital of USTC, Division of Life Sciences and Medicine, University of Science and Technology of China, Hefei, Anhui, China.
⁶ School of Life Sciences, Fudan University, Shanghai, China.
⁷ Second Department of Hepatic Surgery, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China.
⁸ Second Department of Biliary Surgery, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China. yongjie_zh@outlook.com.
⁹ International Cooperation Laboratory on Signal Transduction, National Center for Liver Cancer, Ministry of Education Key Laboratory on Signaling Regulation and Targeting Therapy of Liver Cancer, Shanghai Key Laboratory of Hepato-biliary Tumor Biology, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China. fujing-724@163.com.
¹⁰ International Cooperation Laboratory on Signal Transduction, National Center for Liver Cancer, Ministry of Education Key Laboratory on Signaling Regulation and Targeting Therapy of Liver Cancer, Shanghai Key Laboratory of Hepato-biliary Tumor Biology, Eastern Hepatobiliary Surgery Hospital, Second Military Medical University, Shanghai, China. hywangk@vip.sina.com.

^# Contributed equally.

Abstract

Gallbladder carcinoma (GBC) is the most common biliary tract malignancy with the lowest survival rate, primarily arising from chronic inflammation. To better characterize the progression from inflammation to cancer to metastasis, we performed single-cell RNA sequencing across samples of 6 chronic cholecystitis, 12 treatment-naive GBCs, and 6 matched metastases. Benign epithelial cells from inflamed gallbladders displayed resting, immune-regulating, and gastrointestinal metaplastic phenotypes. A small amount of PLA2G2A⁺ epithelial cells with copy number variation were identified from a histologically benign sample. We validated significant overexpression of PLA2G2A across in situ GBCs, together with increased proliferation and cancer stemness in PLA2G2A-overexpressing GBC cells, indicating an important role for PLA2G2A during early carcinogenesis. Malignant epithelial cells displayed pervasive cancer hallmarks and cellular plasticity, differentiating into metaplastic, inflammatory, and mesenchymal subtypes with distinct transcriptomic, genomic, and prognostic patterns. Chronic cholecystitis led to an adapted microenvironment characterized by MDSC-like macrophages, CD8⁺ T_RM cells, and CCL2⁺ immunity-regulating fibroblasts. By contrast, GBC instigated an aggressive and immunosuppressive microenvironment, featured by tumor-associated macrophages, Treg cells, CD8⁺ T_EX cells, and STMN1⁺ tumor-promoting fibroblasts. Single-cell and bulk RNA-seq profiles consistently showed a more suppressive immune milieu for GBCs with inflammatory epithelial signatures, coupled with strengthened epithelial-immune crosstalk. We further pinpointed a subset of senescence-like fibroblasts (FN1⁺TGM2⁺) preferentially enriched in metastatic lesions, which promoted GBC migration and invasion via their secretory phenotype. Collectively, this study provides comprehensive insights into epithelial and microenvironmental reprogramming throughout cholecystitis-propelled carcinogenesis and metastasis, laying a new foundation for the precision therapy of GBC.

Abstract

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