Abstract
The existence of heterogeneity has plunged cancer treatment into a challenging dilemma. We profiled malignant epithelial cells from 5 gastric adenocarcinoma patients through single-cell sequencing (scRNA-seq) analysis, demonstrating the heterogeneity of gastric adenocarcinoma (GA), and identified the CCKBR+ stem cell-like cancer cells associated poorly differentiated and worse prognosis. We further conducted targeted analysis using single-cell transcriptome libraries, including 40 samples, to confirm these screening results. In addition, we revealed that FOXOs are involved in the progression and development of CCKBR+ gastric adenocarcinoma. Inhibited the expression of FOXOs and disrupting cancer cell stemness reduce the CCKBR+ GA organoid formation and impede tumor progression. Mechanically, CUT&Tag sequencing and Lectin pulldown revealed that FOXOs can activate ST3GAL3/4/5 as well as ST6GALNAC6, promoting elevated sialyation levels in CCKBR+ tumor cells. This FOXO-sialyltransferase axis contributes to the maintenance of homeostasis and the growth of CCKBR+ tumor cells. This insight provides novel perspectives for developing targeted therapeutic strategies aimed at the treating CCKBR associated gastric cancer.
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Data availability
All data used in this study are publicly available as described in the Method section. The web links or unique identifiers for public cohorts/datasets are described in the paper. Source codes and all supplementary data used to generate the results will be deposited once the article is received. All data generated from this study are available without any restrictions, and all raw data of scRNA-seq generated in this study have been deposited at the GEO Datasets (GSE264203). The raw data that support the findings of this study were obtained from Gene Expression Omnibus (GEO) at GSE183904, GSE184613 and TCGA STAD project (https://gdc.cancer.gov).
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Acknowledgements
We express gratitude to all members of the Pathophysiology Department at Anhui Medical University for their valuable comments and advice. Special thanks are extended to Wenyang Ding and Bing Li from OE Biotech Co., Ltd (Shanghai, China) for generously providing single-cell RNA-seq data and offering assistance with bioinformatics analysis. Additionally, we acknowledge the Center for Scientific Research at Anhui Medical University for their invaluable support in conducting our experiments.
Funding
This work was supported by the National Natural Science Foundation of China (reference numbers 82173004, 81870085,81670097 to HZ), Chinese scholarship council Research Collaboration Project (to HZ), Natural Science Foundation of China Hunan Science & Technology Department (2019JJ40429, 2023JJ60428 to LD), Grants for Scientific Research Enhancement of Anhui Medical University (2019xkjT004 and XJ2020019 to HZ), Grants for Collaborative Innovation Project of Colleges and Universities in Anhui Province (GXXT-2021-063 to HZ) and Grants for Postgraduate Scientific Research of Universities in Anhui Province (YJS20210259 to HZ).
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All coauthors have contributed to this study. ZT designed the experiments, analyzed data and wrote this paper. KP and MS were responsible for biopsies collection and data collection. KL and XL interpretated scRNA-seq data. XP was responsible for pathological assessment. ZC, ZY, XY, JZ, JZ, YL, LC, WL, and HM were responsible for biological experiments and manuscript revision. HZ, LD and CK were response for study concept and design, data interpretation and revision of the manuscript.
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The study received approval from the ethics committee of Anhui Medical University and Second Xiangya Hospital (LLSC20210872, LLSC20210594). Written informed consent was obtained from all study participants. Animal experiments were approved by the Animal Research Ethics Committee of Anhui Medical University (LLSC20210972) and complied with the relevant ethical guidelines.
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Tan, Z., Pan, K., Sun, M. et al. CCKBR+ cancer cells contribute to the intratumor heterogeneity of gastric cancer and confer sensitivity to FOXO inhibition. Cell Death Differ 31, 1302–1317 (2024). https://doi.org/10.1038/s41418-024-01360-z
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DOI: https://doi.org/10.1038/s41418-024-01360-z
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