Spatial transcriptomics and multi-omics reveal relapse and resistance mechanisms of EndMT-derived CAFs mediated by TNC and FLNC in glioblastoma

Abstract Background Chemoresistance and recurrence following treatment are the greatest impediments to the prognosis of glioblastoma (GBM). Increasing evidence indicates that cancer-associated fibroblasts (CAFs) play a significant role in the progression of glioblastoma. Nevertheless, the role and s...

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Main Authors: Li Ji, Die Xia, Yu Zhou, Yaling Hu, Zhenkun Yang, Ying Yin, Jingjing Wang, Bo Zhang, Lingli Gong, Koukou Li, Jian Zou, Mei Wang
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Journal of Translational Medicine
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Online Access:https://doi.org/10.1186/s12967-025-06743-5
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author Li Ji
Die Xia
Yu Zhou
Yaling Hu
Zhenkun Yang
Ying Yin
Jingjing Wang
Bo Zhang
Lingli Gong
Koukou Li
Jian Zou
Mei Wang
author_facet Li Ji
Die Xia
Yu Zhou
Yaling Hu
Zhenkun Yang
Ying Yin
Jingjing Wang
Bo Zhang
Lingli Gong
Koukou Li
Jian Zou
Mei Wang
author_sort Li Ji
collection DOAJ
description Abstract Background Chemoresistance and recurrence following treatment are the greatest impediments to the prognosis of glioblastoma (GBM). Increasing evidence indicates that cancer-associated fibroblasts (CAFs) play a significant role in the progression of glioblastoma. Nevertheless, the role and source of CAFs in recurrent and chemotherapy-resistant GBMs still remain ambiguous. Methods Spatial transcriptome (ST) sequencing was conducted on the tissue microarray encompassing primary and recurrent glioma samples in order to characterize the cellular composition. Subsequently, the infiltration of CAFs in our formerly established in vivo temozolomide (TMZ)-resistant model was inspected through immunohistochemical staining. Additionally, we carried out RNA-seq and label-free quantitation (LFQ) proteomics on HCMECs co-cultured with TMZ-sensitive (TMZ-S) or TMZ-resistant (TMZ-R) cells to explore the mechanism. Results This investigation revealed that CAFs and astrocytes are enriched in recurrent GBM, and this phenotype is associated with the expression of extracellular matrix (ECM) proteins associated with COL1A1 and FN1 deposition. Further investigations revealed that tenascin-C (TNC) and filamin C (FLNC), which potentially mediate endothelial-to-mesenchymal transition (EndMT), are the predominant factors that induce the deposition of ECM proteins in the resistance-promoting microenvironment. Additionally, the natural product punicalin (PNC) was found to downregulate EndMT-related proteins, multidrug resistance-associated membrane proteins, and collagen-related proteins by targeting TNC and FLNC, thereby increasing the susceptibility of temozolomide (TMZ)-resistant cells to chemotherapeutic agents both in vitro and in vivo. Conclusion These discoveries indicate that TNC and FLNC induced EndMT was a key resource of CAFs and targeting TNC and FLNC to inhibit EndMT and the collagen pathway is a promising tactic for reversing drug resistance in tumours. The development of combined chemotherapeutic strategies based on the features of tumour microenvironment endothelial cells and ECM deposition has high potential clinical value in increasing the efficacy of tumour treatment.
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spelling doaj-art-6370565319ca4947beb73b3bb7f68e4c2025-08-20T03:04:07ZengBMCJournal of Translational Medicine1479-58762025-07-0123111610.1186/s12967-025-06743-5Spatial transcriptomics and multi-omics reveal relapse and resistance mechanisms of EndMT-derived CAFs mediated by TNC and FLNC in glioblastomaLi Ji0Die Xia1Yu Zhou2Yaling Hu3Zhenkun Yang4Ying Yin5Jingjing Wang6Bo Zhang7Lingli Gong8Koukou Li9Jian Zou10Mei Wang11Center of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityDepartment of Laboratory Medicine, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityCenter of Clinical Research, Wuxi People’s Hospital, Wuxi Medical Center, The Affiliated Wuxi People’s Hospital of Nanjing Medical University, Nanjing Medical UniversityAbstract Background Chemoresistance and recurrence following treatment are the greatest impediments to the prognosis of glioblastoma (GBM). Increasing evidence indicates that cancer-associated fibroblasts (CAFs) play a significant role in the progression of glioblastoma. Nevertheless, the role and source of CAFs in recurrent and chemotherapy-resistant GBMs still remain ambiguous. Methods Spatial transcriptome (ST) sequencing was conducted on the tissue microarray encompassing primary and recurrent glioma samples in order to characterize the cellular composition. Subsequently, the infiltration of CAFs in our formerly established in vivo temozolomide (TMZ)-resistant model was inspected through immunohistochemical staining. Additionally, we carried out RNA-seq and label-free quantitation (LFQ) proteomics on HCMECs co-cultured with TMZ-sensitive (TMZ-S) or TMZ-resistant (TMZ-R) cells to explore the mechanism. Results This investigation revealed that CAFs and astrocytes are enriched in recurrent GBM, and this phenotype is associated with the expression of extracellular matrix (ECM) proteins associated with COL1A1 and FN1 deposition. Further investigations revealed that tenascin-C (TNC) and filamin C (FLNC), which potentially mediate endothelial-to-mesenchymal transition (EndMT), are the predominant factors that induce the deposition of ECM proteins in the resistance-promoting microenvironment. Additionally, the natural product punicalin (PNC) was found to downregulate EndMT-related proteins, multidrug resistance-associated membrane proteins, and collagen-related proteins by targeting TNC and FLNC, thereby increasing the susceptibility of temozolomide (TMZ)-resistant cells to chemotherapeutic agents both in vitro and in vivo. Conclusion These discoveries indicate that TNC and FLNC induced EndMT was a key resource of CAFs and targeting TNC and FLNC to inhibit EndMT and the collagen pathway is a promising tactic for reversing drug resistance in tumours. The development of combined chemotherapeutic strategies based on the features of tumour microenvironment endothelial cells and ECM deposition has high potential clinical value in increasing the efficacy of tumour treatment.https://doi.org/10.1186/s12967-025-06743-5GlioblastomaTMZ resistanceEndMTTenascin-CFilamin C
spellingShingle Li Ji
Die Xia
Yu Zhou
Yaling Hu
Zhenkun Yang
Ying Yin
Jingjing Wang
Bo Zhang
Lingli Gong
Koukou Li
Jian Zou
Mei Wang
Spatial transcriptomics and multi-omics reveal relapse and resistance mechanisms of EndMT-derived CAFs mediated by TNC and FLNC in glioblastoma
Journal of Translational Medicine
Glioblastoma
TMZ resistance
EndMT
Tenascin-C
Filamin C
title Spatial transcriptomics and multi-omics reveal relapse and resistance mechanisms of EndMT-derived CAFs mediated by TNC and FLNC in glioblastoma
title_full Spatial transcriptomics and multi-omics reveal relapse and resistance mechanisms of EndMT-derived CAFs mediated by TNC and FLNC in glioblastoma
title_fullStr Spatial transcriptomics and multi-omics reveal relapse and resistance mechanisms of EndMT-derived CAFs mediated by TNC and FLNC in glioblastoma
title_full_unstemmed Spatial transcriptomics and multi-omics reveal relapse and resistance mechanisms of EndMT-derived CAFs mediated by TNC and FLNC in glioblastoma
title_short Spatial transcriptomics and multi-omics reveal relapse and resistance mechanisms of EndMT-derived CAFs mediated by TNC and FLNC in glioblastoma
title_sort spatial transcriptomics and multi omics reveal relapse and resistance mechanisms of endmt derived cafs mediated by tnc and flnc in glioblastoma
topic Glioblastoma
TMZ resistance
EndMT
Tenascin-C
Filamin C
url https://doi.org/10.1186/s12967-025-06743-5
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