Deciphering the tumor immune microenvironment: single-cell and spatial transcriptomic insights into cervical cancer fibroblasts
Abstract Background Cervical cancer (CC) remains a significant global health challenge despite advancements in screening, HPV vaccination, and therapeutic strategies. Tumor heterogeneity, driven by epigenetic modifications, affects immune evasion, metastasis, and treatment response. Cancer-associate...
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BMC
2025-07-01
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| Series: | Journal of Experimental & Clinical Cancer Research |
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| Online Access: | https://doi.org/10.1186/s13046-025-03432-5 |
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| author | Zhiheng Lin Youwei Zhou Zhenran Liu Wenyang Nie Hengjie Cao Shengnan Li Xuanling Li Lijun Zhu Guangyao Lin Yanyu Ding Yi Jiang Zuxi Gu Lianwei Xu Zhijie Zhao Huabao Cai |
| author_facet | Zhiheng Lin Youwei Zhou Zhenran Liu Wenyang Nie Hengjie Cao Shengnan Li Xuanling Li Lijun Zhu Guangyao Lin Yanyu Ding Yi Jiang Zuxi Gu Lianwei Xu Zhijie Zhao Huabao Cai |
| author_sort | Zhiheng Lin |
| collection | DOAJ |
| description | Abstract Background Cervical cancer (CC) remains a significant global health challenge despite advancements in screening, HPV vaccination, and therapeutic strategies. Tumor heterogeneity, driven by epigenetic modifications, affects immune evasion, metastasis, and treatment response. Cancer-associated fibroblasts (CAFs) play a crucial role in CC progression and therapy resistance. Single-cell sequencing offers new insights but remains underutilized in CC research. This study integrates single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, and deconvolution analysis to identify key genes and immunotherapy targets. By constructing a prognostic model and exploring the immune microenvironment, we aim to provide novel insights into CC pathogenesis and potential therapeutic strategies. Methods We utilized scRNA-seq, spatial transcriptomics, deconvolution analysis, and pseudotime trajectory mapping to delineate fibroblast subtypes within the tumor immune microenvironment (TIME) of CC. Functional annotations, differential gene expression profiling, cell–cell communication pathways, and transcription factor networks were systematically analyzed. A prognostic model based on bulk RNA-seq data was constructed and validated through survival analysis, with correlations to immune microenvironment characteristics. Functional experiments investigated the role of SDC1, a critical mediator of fibroblast-tumor crosstalk. Additionally, Fibroblast–tumor cell co-culture systems and functional assays were employed to investigate the paracrine role of SDC1. The CAF MYH11⁺ subpopulation was isolated via fluorescence-activated cell sorting (FACS). Multiplex immunofluorescence and immunohistochemical analyses were performed on both cultured cells and human cervical cancer tissue samples to characterize the spatial distribution and dynamic remodeling of MYH11 during stromal reorganization. Results Six distinct fibroblast subtypes were identified, including the C0 MYH11 + fibroblasts, which exhibited unique roles in stemness maintenance, metabolic activity, and immune regulation. Spatial and functional analyses revealed that the C0 subtype is central to tumor-fibroblast interactions, particularly through the MDK-SDC1 signaling axis. The prognostic model incorporating fibroblast-specific markers demonstrated robust predictive power for patient survival outcomes. Additionally, in vitro SDC1 knockdown significantly inhibited CC cell proliferation, migration, and invasion. Fibroblasts show spatially regulated heterogeneity, with activation markers enriched in the tumor zone and MYH11 highest in normal zones, indicating dynamic stromal remodeling. C0 MYH11 + CAF Promotes Tumor Cell Proliferation, Migration, and Inhibits Apoptosis via Soluble SDC1. Conclusion Our results illustrate, in some ways, the possible immunomodulatory and tumor supporting roles of CAFs in CC TIME and highlight the possibility that the MDK-SDC1 pathway is a promising therapeutic target. This study not only promotes a partially new understanding of temporal heterogeneity in CC, but also provides a possible reference base for the development of new biomarkers and immunotherapy approaches to improve clinical outcomes. |
| format | Article |
| id | doaj-art-e02f7546656b49dfb9e7140086ec67d1 |
| institution | Kabale University |
| issn | 1756-9966 |
| language | English |
| publishDate | 2025-07-01 |
| publisher | BMC |
| record_format | Article |
| series | Journal of Experimental & Clinical Cancer Research |
| spelling | doaj-art-e02f7546656b49dfb9e7140086ec67d12025-08-20T03:45:39ZengBMCJournal of Experimental & Clinical Cancer Research1756-99662025-07-0144113310.1186/s13046-025-03432-5Deciphering the tumor immune microenvironment: single-cell and spatial transcriptomic insights into cervical cancer fibroblastsZhiheng Lin0Youwei Zhou1Zhenran Liu2Wenyang Nie3Hengjie Cao4Shengnan Li5Xuanling Li6Lijun Zhu7Guangyao Lin8Yanyu Ding9Yi Jiang10Zuxi Gu11Lianwei Xu12Zhijie Zhao13Huabao Cai14Department of Gynecology, Longhua Hospital, Shanghai University of Traditional Chinese MedicineDepartment of Obstetrics and Gynecology, the First Affiliated Hospital of Anhui Medical UniversityDepartment of Obstetrics and Gynecology, the First Affiliated Hospital of Anhui Medical UniversityThe First Clinical Medical College, Shandong University of Traditional Chinese MedicineDepartment of Gynecology, Longhua Hospital, Shanghai University of Traditional Chinese MedicineDepartment of Gynecology, Longhua Hospital, Shanghai University of Traditional Chinese MedicineDepartment of Gynecology, Longhua Hospital, Shanghai University of Traditional Chinese MedicineDepartment of Gynecology, Longhua Hospital, Shanghai University of Traditional Chinese MedicineDepartment of Gynecology, Longhua Hospital, Shanghai University of Traditional Chinese MedicineDepartment of Immunology, School of Basic Medical Sciences, Center for Big Dataand , Population Health of IHM, Anhui Medical UniversityExperiment Center for Science and Technology, Shanghai University of Traditional Chinese MedicineDepartment of Laboratory Animal, School of Experimental Center of Science and Technology, Shanghai University of Traditional Chinese MedicineDepartment of Gynecology, Longhua Hospital, Shanghai University of Traditional Chinese MedicineDepartment of Plastic and Reconstructive Surgery, School of Medicine, Shanghai Ninth People’ S Hospital, Shanghai Jiao Tong UniversityDepartment of Neurosurgery, The First Affiliated Hospital of Anhui Medical UniversityAbstract Background Cervical cancer (CC) remains a significant global health challenge despite advancements in screening, HPV vaccination, and therapeutic strategies. Tumor heterogeneity, driven by epigenetic modifications, affects immune evasion, metastasis, and treatment response. Cancer-associated fibroblasts (CAFs) play a crucial role in CC progression and therapy resistance. Single-cell sequencing offers new insights but remains underutilized in CC research. This study integrates single-cell RNA sequencing (scRNA-seq), spatial transcriptomics, and deconvolution analysis to identify key genes and immunotherapy targets. By constructing a prognostic model and exploring the immune microenvironment, we aim to provide novel insights into CC pathogenesis and potential therapeutic strategies. Methods We utilized scRNA-seq, spatial transcriptomics, deconvolution analysis, and pseudotime trajectory mapping to delineate fibroblast subtypes within the tumor immune microenvironment (TIME) of CC. Functional annotations, differential gene expression profiling, cell–cell communication pathways, and transcription factor networks were systematically analyzed. A prognostic model based on bulk RNA-seq data was constructed and validated through survival analysis, with correlations to immune microenvironment characteristics. Functional experiments investigated the role of SDC1, a critical mediator of fibroblast-tumor crosstalk. Additionally, Fibroblast–tumor cell co-culture systems and functional assays were employed to investigate the paracrine role of SDC1. The CAF MYH11⁺ subpopulation was isolated via fluorescence-activated cell sorting (FACS). Multiplex immunofluorescence and immunohistochemical analyses were performed on both cultured cells and human cervical cancer tissue samples to characterize the spatial distribution and dynamic remodeling of MYH11 during stromal reorganization. Results Six distinct fibroblast subtypes were identified, including the C0 MYH11 + fibroblasts, which exhibited unique roles in stemness maintenance, metabolic activity, and immune regulation. Spatial and functional analyses revealed that the C0 subtype is central to tumor-fibroblast interactions, particularly through the MDK-SDC1 signaling axis. The prognostic model incorporating fibroblast-specific markers demonstrated robust predictive power for patient survival outcomes. Additionally, in vitro SDC1 knockdown significantly inhibited CC cell proliferation, migration, and invasion. Fibroblasts show spatially regulated heterogeneity, with activation markers enriched in the tumor zone and MYH11 highest in normal zones, indicating dynamic stromal remodeling. C0 MYH11 + CAF Promotes Tumor Cell Proliferation, Migration, and Inhibits Apoptosis via Soluble SDC1. Conclusion Our results illustrate, in some ways, the possible immunomodulatory and tumor supporting roles of CAFs in CC TIME and highlight the possibility that the MDK-SDC1 pathway is a promising therapeutic target. This study not only promotes a partially new understanding of temporal heterogeneity in CC, but also provides a possible reference base for the development of new biomarkers and immunotherapy approaches to improve clinical outcomes.https://doi.org/10.1186/s13046-025-03432-5Cervical cancerTumor immune microenvironmentSingle-cell RNA sequencingSpatial transcriptomicsCancer-associated fibroblastsSDC1 |
| spellingShingle | Zhiheng Lin Youwei Zhou Zhenran Liu Wenyang Nie Hengjie Cao Shengnan Li Xuanling Li Lijun Zhu Guangyao Lin Yanyu Ding Yi Jiang Zuxi Gu Lianwei Xu Zhijie Zhao Huabao Cai Deciphering the tumor immune microenvironment: single-cell and spatial transcriptomic insights into cervical cancer fibroblasts Journal of Experimental & Clinical Cancer Research Cervical cancer Tumor immune microenvironment Single-cell RNA sequencing Spatial transcriptomics Cancer-associated fibroblasts SDC1 |
| title | Deciphering the tumor immune microenvironment: single-cell and spatial transcriptomic insights into cervical cancer fibroblasts |
| title_full | Deciphering the tumor immune microenvironment: single-cell and spatial transcriptomic insights into cervical cancer fibroblasts |
| title_fullStr | Deciphering the tumor immune microenvironment: single-cell and spatial transcriptomic insights into cervical cancer fibroblasts |
| title_full_unstemmed | Deciphering the tumor immune microenvironment: single-cell and spatial transcriptomic insights into cervical cancer fibroblasts |
| title_short | Deciphering the tumor immune microenvironment: single-cell and spatial transcriptomic insights into cervical cancer fibroblasts |
| title_sort | deciphering the tumor immune microenvironment single cell and spatial transcriptomic insights into cervical cancer fibroblasts |
| topic | Cervical cancer Tumor immune microenvironment Single-cell RNA sequencing Spatial transcriptomics Cancer-associated fibroblasts SDC1 |
| url | https://doi.org/10.1186/s13046-025-03432-5 |
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