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Fibroblast Activation Protein (FAP)+ cancer-associated fibroblasts induce macrophage M2-like polarization via the Fibronectin 1-Integrin α5β1 axis in breast cancer

Oncogene volume 44, pages 2396–2412 (2025)Cite this article

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Abstract

Cancer-associated fibroblasts expressing fibroblast activation protein (FAP+ CAFs) are critical modulators of the breast cancer microenvironment, yet their immunoregulatory mechanisms remain poorly understood. Through integrated analysis of single-cell RNA sequencing data, clinical specimens, and in vivo and in vitro experiments, we identified FAP+ CAFs as the predominant stromal population associated with poor clinical outcomes and immunosuppressive features. Mechanistically, FAP+ CAFs secrete high levels of fibronectin 1 (FN1), which engages integrin α5β1 on macrophages to trigger FAK-AKT-STAT3 signaling, driving their polarization toward an immunosuppressive M2-like phenotype. Importantly, pharmacological disruption of FN1-integrin α5β1 signaling using Cilengitide effectively reprogrammed the tumor immune landscape and suppressed tumor growth in mice models. These findings establish FAP+ CAF-derived FN1 as a critical orchestrator of tumor immunosuppression and identify the FN1-integrin α5β1 axis as a promising therapeutic target in breast cancer.

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Fig. 1: Characteristics and implications of FAP+ CAFs in breast cancer microenvironment.
Fig. 2: Association of high FN1 expression with an immunosuppressive microenvironment in breast cancer.
Fig. 3: FAP+ CAFs promote macrophage M2-like polarization and suppress T cell responses.
Fig. 4: FN1 induced macrophage M2-like polarization through integrin α5β1 signaling.
Fig. 5: Integrin α5β1 blockade reverses FN1-induced immunosuppressive macrophage phenotype and function.
Fig. 6: FN1 promotes macrophage M2-like polarization through the FAK-AKT-STAT3 signaling cascade.
Fig. 7: FN1 and Cilengitide competitively regulate tumor growth and macrophage polarization through integrin α5β1.
Fig. 8: Spatial characterization of FN1-modulated tumor immune microenvironment.

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Data availability

The single-cell RNA sequencing data have been deposited in the GSA-Human repository (https://ngdc.cncb.ac.cn/gsa-human/browse/HRA003664). The processed data and analysis results are available within the article and its supplementary files. Other raw data supporting this study are available in the figshare repository (DOI: 10.6084/m9.figshare.26648908). Additional data of this study are also available from the corresponding authors upon reasonable request.

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Acknowledgements

The authors wish to express their gratitude to Dr. Jun Ye, Dr. Ke Wang, and Dr. Pin Wu for their invaluable advice and insights during the preparation of this manuscript. Their expertise significantly contributed to the completion of this work.

Funding

This work was supported by the National Natural Science Foundation of China (82403697 to W.Z.C., 82273275 to C.N.), the Natural Science Foundation of Zhejiang Province (ZCLY24H1601 to W.Z.C., LRG25H160001 to C.N.), the Medical and Health Science and Technology Project of Zhejiang Province (2023KY861 to M.J.J., 2023KY046 to W.J.X.), the Zhejiang Province Traditional Chinese Medicine Science and Technology Project (2023ZR096 to X.B.Z.), and the National Key Research and Development Program of China (2022YFC2505100 to W.Z.C., 2022YFA1105200 to J.H.).

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Author notes

  1. These authors contributed equally: Wuzhen Chen, Mengjie Jiang, Xinbo Zou.

Authors and Affiliations

  1. Department of Breast Surgery, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China

    Wuzhen Chen, Zhigang Chen, Lesang Shen, Jianming Hu, Jian Huang & Chao Ni

  2. Key Laboratory of Tumor Microenvironment and Immune Therapy of Zhejiang Province, Second Affiliated Hospital, Zhejiang University School of Medicine, Hangzhou, Zhejiang, China

    Wuzhen Chen, Zhigang Chen, Lesang Shen, Jianming Hu, Jian Huang & Chao Ni

  3. Department of Oncology, Lanxi People’s Hospital, Jinhua, Zhejiang, China

    Wuzhen Chen

  4. Department of Radiotherapy, First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Traditional Chinese Medicine), Hangzhou, Zhejiang, China

    Mengjie Jiang

  5. Department of Otolaryngology, First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Traditional Chinese Medicine), Hangzhou, Zhejiang, China

    Xinbo Zou

  6. Suzhou Medical College of Soochow University, Suzhou, Jiangsu, China

    Mingxiang Kong

  7. Cancer Center, Zhejiang University, Hangzhou, China

    Jian Huang & Chao Ni

  8. General Surgery, Cancer Center, Department of Breast Surgery, Zhejiang Provincial People’s Hospital (Affiliated People’s Hospital, Hangzhou Medical College), Hangzhou, Zhejiang, China

    Wenjie Xia

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Contributions

WZC and WJX designed the study, supervised the project, and drafted the manuscript. MJJperformed the bioinformatics analysis, managed data interpretation, and participated in manuscript revision. LSS, XBZ, MXK, and JMH conducted the in vitro and in vivo experiments respectively, contributing to data collection and analysis. ZGC, JH, and CN focused on revising the manuscript for critical content. All authors reviewed, edited, and approved the final manuscript, ensuring the accuracy and integrity of the work.

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Correspondence to Jian Huang, Chao Ni or Wenjie Xia.

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This study received ethical approval from the Ethical Committee of the Second Affiliated Hospital, Zhejiang University School of Medicine (Approval No. YAN 2021-0421, June 2021). Written informed consent was obtained from all participants providing clinical samples. All procedures were conducted in strict accordance with the guidelines and regulations approved by the Ethical Committee.

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Chen, W., Jiang, M., Zou, X. et al. Fibroblast Activation Protein (FAP)+ cancer-associated fibroblasts induce macrophage M2-like polarization via the Fibronectin 1-Integrin α5β1 axis in breast cancer. Oncogene 44, 2396–2412 (2025). https://doi.org/10.1038/s41388-025-03359-3

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