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Translational Therapeutics

Heterogeneity of tertiary lymphoid structures predicts the response to neoadjuvant therapy and immune microenvironment characteristics in triple-negative breast cancer

British Journal of Cancer volume 132, pages 295–310 (2025)Cite this article

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Abstract

Background

Tertiary lymphoid structures (TLSs) impact cancer outcomes, including in triple-negative breast cancer (TNBC), where their role in immune modulation during neoadjuvant therapy (NAT) is underexplored.

Methods

This study employed single-cell RNA sequencing (scRNA-seq), multiplex immunofluorescence (mIF) staining, and radiomic techniques to evaluate TLSs and the tumour microenvironment (TME) in TNBC patient samples before and after NAT.

Results

The presence of TLSs in TNBC was associated with B-cell maturation and T-cell activation. Compared with TLS-low TNBC, TLS-high TNBC showed significantly greater expression of immunoglobulin family genes (IGHM and IGHG1) in B cells and greater cytotoxicity of neoantigen-specific CD8 + T cells (neoTCR8). Additionally, mIF revealed notable differences between TLSs and the TME in TNBC. Although CD8 + T-cell levels do not predict the NAT response effectively, TLS maturity strongly correlated with better NAT outcomes and prognosis (P < 0.05). An imaging biomarker scoring system was also developed to predict TLS status and NAT efficacy.

Conclusion

Our results demonstrated changes in TLSs and the TME in TNBC patients post-NAT. These findings confirm the predictive value of mature TLSs (mTLSs) and support the use of personalised immunotherapy based on post-NAT immune characteristics, thereby improving clinical outcomes.

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Fig. 1: Impact of TLSs on BC Prognosis.
Fig. 2: Functional roles of B Cells within TLSs in TNBC.
Fig. 3: Functional T cells within TLSs in TNBC.
Fig. 4: CNV disruption in TLS formation.
Fig. 5: Identification of TLSs in BC.
Fig. 6: Assessing the heterogeneity of TLS and TME using mIF.
Fig. 7: Predictive model of TLSs using DCE-MRI radiomics and machine learning.

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Data availability

The data are available upon reasonable request. All data relevant to this study are included in the article or uploaded as online supplemental information.

Code availability

The underlying code for this study, along with the associated training and validation datasets, is not publicly available. However, it can be made available to qualified researchers upon a reasonable request to the corresponding author.

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Acknowledgements

We would like to thank the staff members of the TCGA and GEO Research Network, as well as all of the authors, for making their valuable research data public.

Funding

The present study was supported by the High-level Talent Introduction Project of Fujian Cancer Hospital (No.: F2328R-GC301-01) and the High-level Talent Training Program of Fujian Cancer Hospital (No.: 2024YNG03).

Author information

Author notes

  1. These authors contributed equally: Qing Wang, Yushuai Yu, Chenxi Wang.

Authors and Affiliations

  1. Department of Breast Surgery, Clinical Oncology School of Fujian Medical University, Fujian Cancer Hospital, Fuzhou, Fujian, 350014, China

    Qing Wang, Yushuai Yu, Zirong Jiang, Xiaofen Li, Xiewei Huang & Chuangui Song

  2. Department of Breast Surgery, Yunnan Cancer Hospital, The Third Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, 650118, China

    Chenxi Wang, Ying Song & Shicong Tang

  3. Rehabilitation College, Kunming Medical University, Kunming, Yunnan, 650118, China

    Jialu Li

  4. Department of Radiology, Yunnan Cancer Hospital, The Third Affiliated Hospital of Kunming Medical University, Kunming, Yunnan, 650118, China

    Zhenhui Li

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Contributions

CGS, SCT and ZHL supported and designed the study. QW, XWH, XFL, YS, and JLL collected, processed, and analysed the datasets. CXW, ZRJ, and XWH verified the data. QW and YSY wrote the manuscript. All of the authors have read and approved the final version of the manuscript.

Corresponding authors

Correspondence to Zhenhui Li, Shicong Tang or Chuangui Song.

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The authors declare no competing interests.

Ethical approval

This study was approved by the Ethics Committee of Yunnan Provincial Cancer Hospital (Approval No. KYLX2024-060) and Fujian Cancer Hospital (Approval No. K2024-056-01), and written informed consent was obtained from all participants.

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Wang, Q., Yu, Y., Wang, C. et al. Heterogeneity of tertiary lymphoid structures predicts the response to neoadjuvant therapy and immune microenvironment characteristics in triple-negative breast cancer. Br J Cancer 132, 295–310 (2025). https://doi.org/10.1038/s41416-024-02917-y

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