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Cytokine CCL2 secreted by cancer-associated fibroblasts augments temozolomide resistance in glioblastoma through ERK1/2 signaling

Oncogene volume 44, pages 4657–4670 (2025)Cite this article

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Abstract

The intricate tumor microenvironment largely influences chemoresistance in glioblastoma. Cancer-associated fibroblasts (CAFs) that modulate tumor progression have recently been identified as non-tumor stromal cells within the glioblastoma microenvironment. It remains unclear whether CAFs play a role in conferring chemoresistance to glioblastoma. The effects and mechanisms of CAFs on glioblastoma cells under temozolomide (TMZ) treatment are investigated by a series of patient-derived CAFs, orthotopic xenograft mouse models, and glioblastoma organoids (GBOs). Patient-derived cells have a transcriptomic and biomolecular profile of CAFs. CAFs promote temozolomide resistance in glioblastoma in vitro; these findings are consistent with results from intracranial tumor xenografts and GBO models. Mechanistically, CAFs express and secrete a significantly higher C-C motif chemokine ligand 2 (CCL2), which selectively enhances the activation of the ERK1/2 signaling in glioblastoma cells. Pharmacologically disrupting the CCL2-CCR2 axis or MEK1/2-ERK1/2 pathway effectively restores the therapeutic efficacy of temozolomide in glioblastoma cells and patient-derived GBOs. The decreased phosphor-ERK1/2 expression induced by trametinib treatment is also observed in glioblastoma cells following the CCL2-CCR2 axis inhibition. The present study suggests that targeting the CCL2/CCR2/ERK1/2 pathway may help overcome chemoresistance in glioblastomas caused by CAFs.

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Fig. 1: CAFs are associated with worse survival and poor TMZ response for gliomas.
Fig. 2: Primary culture and identification of patient-derived CAFs.
Fig. 3: CAF promotes the TMZ resistance of GBM cells.
Fig. 4: CAF promotes TMZ resistance of GBM cells via cytokine CCL2.
Fig. 5: CCL2-CCR2 signaling activates the ERK1/2 expression in GBM cells to potentiate TMZ resistance.
Fig. 6: Pharmacological inhibition of CCR2 or ERK1/2 enhances TMZ efficacy in patient-derived GBO models.
Fig. 7: Mechanistic model of CAF-mediated chemoresistance in glioblastoma.

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Data availability

Raw sequencing data is available at Genome Sequence Archive for Humans (https://ngdc.cncb.ac.cn/gsa-human/s/OnfgtAhQ) with accession HRA002403 and could be accessed under regulation of the GSA-Human Data Access Agreement. The code used for analyses is available at https://github.com/sxz-ivan/GAF-analysis. Figureshare https://doi.org/10.6084/m9.figshare.28603253. All data are available in the main text or the supplementary materials.

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Acknowledgements

This study is supported by the National Natural Science Foundation of China (82272644 to YL, 82372836 to WY, 82403965 to SC), Sichuan Science and Technology Program grant (2023YFQ0002 to YL), Sichuan Science and Technology Program grant (2023YFSY0042 to WY), Sichuan Science and Technology Program grant (2023YFG0127 to YY), and Sichuan Provincial Foundation of Science and Technology grant (2023NSFSC1867 to SZ).

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  1. These authors contributed equally: Mingrong Zuo, Shuxin Zhang, Siliang Chen.

Authors and Affiliations

  1. Department of Neurosurgery, West China Hospital, Sichuan University, Chengdu, Sichuan Province, China

    Mingrong Zuo, Siliang Chen, Yuze He, Junhong Li, Yufan Xiang, Yunbo Yuan, Wanchun Yang, Zhihao Wang, Wenhao Li, Yuan Yang, Yunhui Zeng, Qing Mao & Yanhui Liu

  2. Department of Pediatric Neurosurgery, West China Second University Hospital, Key Laboratory of Birth Defects and Related Diseases of Women and Children of MOE, Sichuan University, Chengdu, Sichuan Province, China

    Mingrong Zuo & Tengfei Li

  3. Department of Intensive Care Unit, West China Hospital, Sichuan University, No. 37 Guoxue Road, Chengdu, Sichuan Province, China

    Shuxin Zhang

  4. Department of Pathology, West China Hospital, Sichuan University, No. 37 Guoxue Road, Chengdu, Sichuan Province, China

    Ni Chen

  5. Department of Biotherapy, Cancer Center and State Key Laboratory of Biotherapy, West China Hospital, Sichuan University, Chengdu, Sichuan Province, China

    Mina Chen

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Contributions

MRZ, MNC, and YHL investigated and designed the study. MRZ, SXZ, SLC, YBY, YZH, and WHL performed all experiments. MRZ, SXZ, YBY, SLC, WCY, TFL, ZHW, and YFX collected and analyzed all data. NC examined the diagnosis of human gliomas. MRZ, SXZ, MNC, YHZ, YY, QM, and YHL supervised the study. All authors wrote, reviewed, edited, and approved the publication of the original paper.

Corresponding authors

Correspondence to Mina Chen or Yanhui Liu.

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Competing interests

MZ and YL hold an authorized Chinese invention patent relating to the method of the primary CAFs culturing (ZL202111097340.7). Other authors reported no conflict of interest.

Ethics approval and consent to participate

All procedures were performed in accordance with the ethical standards as outlined in the 1964 Declaration of Helsinki and its subsequent amendments, or with comparable ethical standards. All human tissues utilized in the present study were approved by the Ethics Committee on Biomedical Research, West China Hospital of Sichuan University (No.2021.1319), and attained written informed consent from patients or their legal guardians. All animal experiments were approved by the Institutional Animal Care and Use Committee (IACUC) of West China Hospital, Sichuan University, in compliance with the Guide for the Care and Use of Laboratory Animals.

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Zuo, M., Zhang, S., Chen, S. et al. Cytokine CCL2 secreted by cancer-associated fibroblasts augments temozolomide resistance in glioblastoma through ERK1/2 signaling. Oncogene 44, 4657–4670 (2025). https://doi.org/10.1038/s41388-025-03601-y

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