Single-cell RNA sequencing and spatial transcriptomics reveal the heterogeneity and intercellular communication of cancer-associated fibroblasts in gastric cancer

Abstract Background Gastric cancer is a highly aggressive malignancy characterized by a complex tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs), which are a key component of the TME, exhibit significant heterogeneity and play crucial roles in tumor progression. Therefore, a compre...

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Main Authors: Xijie Zhang, Bo Ren, Bo Liu, Rui Wang, Sen Li, Yuzhou Zhao, Wence Zhou
Format: Article
Language:English
Published: BMC 2025-03-01
Series:Journal of Translational Medicine
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Online Access:https://doi.org/10.1186/s12967-025-06376-8
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author Xijie Zhang
Bo Ren
Bo Liu
Rui Wang
Sen Li
Yuzhou Zhao
Wence Zhou
author_facet Xijie Zhang
Bo Ren
Bo Liu
Rui Wang
Sen Li
Yuzhou Zhao
Wence Zhou
author_sort Xijie Zhang
collection DOAJ
description Abstract Background Gastric cancer is a highly aggressive malignancy characterized by a complex tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs), which are a key component of the TME, exhibit significant heterogeneity and play crucial roles in tumor progression. Therefore, a comprehensive understanding of CAFs is essential for developing novel therapeutic strategies for gastric cancer. Methods This study investigates the characteristics and functional information of CAF subtypes and explores the intercellular communication between CAFs and malignant epithelial cells (ECs) in gastric cancer by analyzing single-cell sequencing data from 24 gastric cancer samples. CellChat was employed to map intercellular communication, and Seurat was used to integrate single-cell sequencing data with spatial transcriptome data to reconstruct a comprehensive single-cell spatial map. The spatial relationship between apCAFs and cancer cells was analyzed using multicolor immunohistochemistry. Results Cells were categorized into nine distinct categories, revealing a positive correlation between the proportions of epithelial cells (ECs) and fibroblasts. Furthermore, six fibroblast subpopulations were identified: inflammatory (iCAFs), pericytes, matrix (mCAFs), antigen-presenting (apCAFs), smooth muscle cells (SMCs), and proliferative CAFs (pCAFs). Each of these subpopulations was linked to various biological processes and immune responses. Malignant ECs exhibited heightened intercellular communication, particularly with CAF subpopulations, through specific ligand-receptor interactions. High-density regions of CAF subpopulations displayed spatial exclusivity, with pericytes serving as a source for iCAFs, mCAFs, and apCAFs. Notably, malignant ECs and apCAFs showed increased interactions, with certain ligand-receptor pairs potentially impacting the prognosis of gastric cancer. Multiplex immunohistochemistry (mIHC) confirmed the close spatial proximity of apCAFs to cancer cells in gastric cancer. Conclusion Our study provided a comprehensive characterization of CAF heterogeneity in gastric cancer and revealed the intricate intercellular networks within the TME. The identified CAF subpopulations and their interactions with malignant cells could serve as potential therapeutic targets.
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spelling doaj-art-a1fa4af80be7407b82b0934ee304ff1c2025-08-20T02:41:34ZengBMCJournal of Translational Medicine1479-58762025-03-0123111410.1186/s12967-025-06376-8Single-cell RNA sequencing and spatial transcriptomics reveal the heterogeneity and intercellular communication of cancer-associated fibroblasts in gastric cancerXijie Zhang0Bo Ren1Bo Liu2Rui Wang3Sen Li4Yuzhou Zhao5Wence Zhou6The Second Clinical Medical School, Lanzhou UniversityThe Second Clinical Medical School, Lanzhou UniversityThe Second Clinical Medical School, Lanzhou UniversityThe Second Clinical Medical School, Lanzhou UniversityDepartment of General Surgery, Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer HospitalDepartment of General Surgery, Affiliated Cancer Hospital of Zhengzhou University & Henan Cancer HospitalThe Second Clinical Medical School, Lanzhou UniversityAbstract Background Gastric cancer is a highly aggressive malignancy characterized by a complex tumor microenvironment (TME). Cancer-associated fibroblasts (CAFs), which are a key component of the TME, exhibit significant heterogeneity and play crucial roles in tumor progression. Therefore, a comprehensive understanding of CAFs is essential for developing novel therapeutic strategies for gastric cancer. Methods This study investigates the characteristics and functional information of CAF subtypes and explores the intercellular communication between CAFs and malignant epithelial cells (ECs) in gastric cancer by analyzing single-cell sequencing data from 24 gastric cancer samples. CellChat was employed to map intercellular communication, and Seurat was used to integrate single-cell sequencing data with spatial transcriptome data to reconstruct a comprehensive single-cell spatial map. The spatial relationship between apCAFs and cancer cells was analyzed using multicolor immunohistochemistry. Results Cells were categorized into nine distinct categories, revealing a positive correlation between the proportions of epithelial cells (ECs) and fibroblasts. Furthermore, six fibroblast subpopulations were identified: inflammatory (iCAFs), pericytes, matrix (mCAFs), antigen-presenting (apCAFs), smooth muscle cells (SMCs), and proliferative CAFs (pCAFs). Each of these subpopulations was linked to various biological processes and immune responses. Malignant ECs exhibited heightened intercellular communication, particularly with CAF subpopulations, through specific ligand-receptor interactions. High-density regions of CAF subpopulations displayed spatial exclusivity, with pericytes serving as a source for iCAFs, mCAFs, and apCAFs. Notably, malignant ECs and apCAFs showed increased interactions, with certain ligand-receptor pairs potentially impacting the prognosis of gastric cancer. Multiplex immunohistochemistry (mIHC) confirmed the close spatial proximity of apCAFs to cancer cells in gastric cancer. Conclusion Our study provided a comprehensive characterization of CAF heterogeneity in gastric cancer and revealed the intricate intercellular networks within the TME. The identified CAF subpopulations and their interactions with malignant cells could serve as potential therapeutic targets.https://doi.org/10.1186/s12967-025-06376-8Gastric cancerCancer-associated fibroblastsSingle-cell transcriptomicsCell–cell interactionsTumor microenvironment
spellingShingle Xijie Zhang
Bo Ren
Bo Liu
Rui Wang
Sen Li
Yuzhou Zhao
Wence Zhou
Single-cell RNA sequencing and spatial transcriptomics reveal the heterogeneity and intercellular communication of cancer-associated fibroblasts in gastric cancer
Journal of Translational Medicine
Gastric cancer
Cancer-associated fibroblasts
Single-cell transcriptomics
Cell–cell interactions
Tumor microenvironment
title Single-cell RNA sequencing and spatial transcriptomics reveal the heterogeneity and intercellular communication of cancer-associated fibroblasts in gastric cancer
title_full Single-cell RNA sequencing and spatial transcriptomics reveal the heterogeneity and intercellular communication of cancer-associated fibroblasts in gastric cancer
title_fullStr Single-cell RNA sequencing and spatial transcriptomics reveal the heterogeneity and intercellular communication of cancer-associated fibroblasts in gastric cancer
title_full_unstemmed Single-cell RNA sequencing and spatial transcriptomics reveal the heterogeneity and intercellular communication of cancer-associated fibroblasts in gastric cancer
title_short Single-cell RNA sequencing and spatial transcriptomics reveal the heterogeneity and intercellular communication of cancer-associated fibroblasts in gastric cancer
title_sort single cell rna sequencing and spatial transcriptomics reveal the heterogeneity and intercellular communication of cancer associated fibroblasts in gastric cancer
topic Gastric cancer
Cancer-associated fibroblasts
Single-cell transcriptomics
Cell–cell interactions
Tumor microenvironment
url https://doi.org/10.1186/s12967-025-06376-8
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