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Tumor-derived exosomes drive pre-metastatic niche formation in lung via modulating CCL1+ fibroblast and CCR8+ Treg cell interactions

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Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Background

Since the lung is one of the most common sites for cancer metastasis, it could provide a suitable microenvironment for pre-metastatic niche (PMN) formation to facilitate tumor cell colonization. Regulatory T cells (Tregs) are an immunosuppressive cell type found ubiquitously in tumors and may play a crucial role in PNM formation. In this study, we investigated tumor-derived exosome (TDE)-induced Treg differentiation in the lung PMN as well as the underlying mechanisms.

Methods

TDEs were isolated from the Lewis lung carcinoma cell line (LLC-exo) and their effects on mouse pulmonary fibroblasts was investigated in vitro as well as on lung tumor formation and metastasis in a pre-injected mouse model. Immune cell populations in the lung were analyzed by flow cytometry. Expression of CCL1 and CCR8 was evaluated by immunofluorescence staining, qRT-PCR and Western blot analyses. Cytokine expression was measured using mouse cytokine arrays and ELISA.

Results

The number of CD4+ FoxP3+ Tregs was significantly increased in lungs in a LLC-exo pre-injected mouse model. Lung fibroblasts secreted increased amounts of CCL1 after co-culture with LLC-exo, which induced Treg differentiation by activating its specific receptor CCR8, ultimately contributing to the establishment of an immunologically tolerant PMN. Moreover, inhibiting the release of LLC-exo by GW4869, or blocking the CCL1-CCR8 axis using AZ084, suppressed Tregs differentiation and tumor metastasis in the lung.

Conclusions

Collectively, our study provides a novel mechanism by which Tregs are activated to form an immunologically tolerant PMN and demonstrates a critical link among lung fibroblasts, Tregs and metastatic tumor cells.

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Availability of data and materials

The authors declare that all data and materials are available on request.

Abbreviations

CAF:

Cancer-associated fibroblasts

DMED:

Dulbecco’s Modified Eagle’s Medium

ECM:

Extracellular matrix

EV:

Extracellular vesicles

FBS:

Fetal bovine serum

HE:

Hematoxylin and eosin

LLC-exo:

Lewis lung carcinoma exosome

MDSCs:

Myeloid-derived suppressor cells

MPF:

Mouse pulmonary fibroblasts

PMN:

Pre-metastatic niche

RBC:

Red blood cells

TDE:

Tumor cell-derived exosomes

TME:

Tumor microenvironment

Tregs:

Regulatory T cells

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Acknowledgements

We thank Dr. Jessica Tamanini from Insight Editing London for critical reading of the manuscript.

Funding

This work was supported by the National Natural Science Foundation of China (Grant Nos. 2021YFA1201102, 82073231) and the Henan Province Scientific and Technological Research (Grant Nos. 222102310728).

Author information

Author notes

  1. Ming Wang and Zhongyu Qin have contributed equally to this work.

Authors and Affiliations

  1. Medical Research Center, The First Affiliated Hospital of Zhengzhou University, Zhengzhou University, Zhengzhou, 450052, Henan, China

    Ming Wang, Jiajia Wan, Yan Yan, **xi Duan, **aohan Yao, Wenqing Li & Zhihai Qin

  2. Changzhi Medical College, Changzhi, 046000, Shanxi, China

    Zhongyu Qin

  3. Department of Urology, The First Affiliated Hospital of Zhengzhou University, Zhengzhou, 450052, Henan, China

    Ziming Jiang

  4. Academy of Medical Sciences, Zhengzhou University, Zhengzhou, 450052, Henan, China

    Zhihai Qin

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  1. Ming Wang
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Contributions

MW and ZHQ designed the project. MW and ZYQ conducted most of the experiments. YY and XXD performed animal experiments. JJW and XHY responsible for immunofluorescence staining. ZMJ, WQL performed some of the cell culture experiments. MW and ZYQ analyzed the data. MW and ZYQ wrote and edited the manuscript. All authors have read and agreed the contents of the manuscript and consent to its publication.

Corresponding authors

Correspondence to Ming Wang or Zhihai Qin.

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Conflict of interest

The authors declare no competing interests.

Ethical approval

All animal experiments were approved by the Ethical Review Committee of the First Affiliated Hospital of Zhengzhou University under approval number 2020-KY-154.

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Wang, M., Qin, Z., Wan, J. et al. Tumor-derived exosomes drive pre-metastatic niche formation in lung via modulating CCL1+ fibroblast and CCR8+ Treg cell interactions. Cancer Immunol Immunother 71, 2717–2730 (2022). https://doi.org/10.1007/s00262-022-03196-3

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