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Tumor-associated macrophages expressing galectin-9 identify immunoevasive subtype muscle-invasive bladder cancer with poor prognosis but favorable adjuvant chemotherapeutic response

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Abstract

Purpose

Tumor-associated macrophages (TAMs) exist as heterogeneous subsets and have dichotomous roles in cancer-immune evasion. This study aims to assess the clinical effects of Galectin-9+ tumor-associated macrophages (Gal-9+TAMs) in muscle-invasive bladder cancer (MIBC).

Experimental design

We identified Gal-9+TAMs by immunohistochemistry (IHC) analysis of a tumor microarray (TMA) (n = 141) from the Zhongshan Hospital and by flow cytometric analysis of tumor specimens (n = 20) from the Shanghai Cancer Center. The survival benefit of platinum-based chemotherapy in this subpopulation was evaluated. The effect of the tumor-immune microenvironment with different percentages of Gal-9+TAMs was explored.

Results

The frequency of Gal-9+TAMs increased with tumor stage and grade. Gal-9+TAMs predicted poor overall survival (OS) and recurrence-free survival (RFS) and were better than Gal-9−TAMs and TAMs to discriminate prognostic groups. In univariate and multivariate Cox regression analyses, patients with high percentages of Gal-9+TAMs showed the prominent survival benefit after receiving adjuvant chemotherapy (ACT). High Gal-9+TAM infiltration correlated with increasing numbers of regulatory T cells (Tregs) and mast cells and decreasing numbers of CD8+T and dendritic cells (DCs). Dense infiltration of Gal-9+TAMs was related to reduced cytotoxic molecules, enhanced immune checkpoints or immunosuppressive cytokines expressed by immune cells, as well as active proliferation of tumor cells. Additionally, the subpopulation accumulated was strongly associated with PD-1+TIM-3+CD8+T cells.

Conclusions

Gal-9+TAMs predicted OS and RFS and response to ACT in MIBC patients. High Gal-9+TAMs were associated with a pro-tumor immune contexture concomitant with T cell exhaustion.

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Abbreviations

ACT:

Adjuvant chemotherapy

AJCC:

American Joint Committee on Cancer

AP:

Alkaline phosphatase

DAB:

Diaminobenzidine

DBSS:

Dulbecco’s balanced salt solution

FCM:

Flow cytometry

Gal-9+TAMs:

Galectin-9+ tumor-associated macrophages

Gal-9−TAMs:

Galectin-9− tumor-associated macrophages

GZMB:

Granzyme B

HCI:

Hydrochloric acid

HR:

Hazard ratio

HRP:

Horseradish peroxidase

ICIs:

Immune checkpoint inhibitors

LVI:

Lymphovascular invasion

MIBC:

Muscle-invasive bladder cancer

PRF1:

Perforin 1

RFS:

Recurrence-free survival

TAMs:

Tumor-associated macrophages

TIGIT:

T-cell Ig and ITIM domain

TMA:

Tissue microarray

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Funding

This study was funded by grants from the National Natural Science Foundation of China (81671628, 31770851, 81702496, 81702497, 81702805, 81772696, 81871306, 81872082, 81902556, 81902563, 81902898, 81974393), the National Key R&D Program of China (2017YFC0114303), the Shanghai Municipal Natural Science Foundation (16ZR1406500, 17ZR1405100, 19ZR1431800), the Guide Project of Science and Technology Commission of Shanghai Municipality (17411963100), the Shanghai Sailing Program (18YF1404500, 19YF1407900, 19YF1427200), the Shanghai Municipal Commission of Health and Family Planning Program (20174Y0042, 201840168, 20184Y0151), the Fudan University Shanghai Cancer Center for Outstanding Youth Scholars Foundation (YJYQ201802) and a grant from the Shanghai Cancer Research Charity Center. None of the study sponsors contributed to the study design, or the collection, analysis or interpretation of data.

Author information

Author notes

  1. Yangyang Qi, Yuan Chang, Zewei Wang and Lingli Chen contributed equally to this work.

Authors and Affiliations

  1. Department of Immunology, School of Basic Medical Sciences, Fudan University, Building West 13, No. 138 Yixueyuan Road, Shanghai, 200032, China

    Yangyang Qi, Yifan Chen & Weijuan Zhang

  2. Department of Urology, Fudan University Shanghai Cancer Center, Shanghai, China

    Yuan Chang, Zheng Liu, Yu Zhu & Bo Dai

  3. Department of Urology, Zhongshan Hospital, Fudan University, Shanghai, China

    Zewei Wang, Jiajun Wang, Qi Bai, Yu Xia, Li Liu & Jianming Guo

  4. Department of Pathology, Zhongshan Hospital, Fudan University, Shanghai, China

    Lingli Chen

  5. Department of Pathology, Fudan University Shanghai Cancer Center, Shanghai, China

    Yunyi Kong

  6. Department of Pathology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Peipei Zhang

  7. Department of Biochemistry and Molecular Biology, School of Basic Medical Sciences, Fudan University, Building West 7, No. 138 Yixueyuan Road, Shanghai, 200032, China

    Quan Zhou & Jiejie Xu

  8. Department of Urology, Ruijin Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai, China

    Le Xu

  9. Department of Urology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, No. 639 Manufacturing Bureau Road, Shanghai, 200011, China

    Yiwei Wang

Authors
  1. Yangyang Qi
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  2. Yuan Chang
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  3. Zewei Wang
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  4. Lingli Chen
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Contributions

YQ, YC, ZW and LC contributed to the acquisition, analysis and interpretation of data, statistical analysis and drafting of the manuscript. YK, PZ, ZL, QZ, YC, JW, QB, YX, LL, YZ, LX, BD and JG provided technical and material support; YW, WZ and JX were responsible for the study concept and design, analysis and interpretation of data, drafting of the manuscript, obtaining funding and study supervision. All authors read and approved the final manuscript.

Corresponding authors

Correspondence to Yiwei Wang, Jiejie Xu or Weijuan Zhang.

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Conflict of interest

The authors declare they have no conflict of interest.

Ethics approval and ethical standards

This study was approved by the Clinical Research Ethics Committee of Zhongshan Hospital, Fudan University (No. B2015-030) and the institutional review board and the ethics committee of Shanghai Cancer Center, Fudan University (No. 050432-4-1212B). This study was performed following the ethical principles of the Helsinki Declaration.

Informed consent

Patients from the Zhongshan hospital and the Shanghai Cancer Center signed informed consent forms before surgery that permitted the usage of specimens and clinical data for research and publication under the condition of anonymity.

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Parts of this paper were presented at the 17th International Congress of Immunology, 19–23 October 2019, Beijing, China, and published as an abstract in the European Journal of Immunology [1].

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Qi, Y., Chang, Y., Wang, Z. et al. Tumor-associated macrophages expressing galectin-9 identify immunoevasive subtype muscle-invasive bladder cancer with poor prognosis but favorable adjuvant chemotherapeutic response. Cancer Immunol Immunother 68, 2067–2080 (2019). https://doi.org/10.1007/s00262-019-02429-2

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