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Cancer Immunology, Immunotherapy

, Volume 68, Issue 12, pp 2067–2080 | Cite as

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

  • Yangyang Qi
  • Yuan Chang
  • Zewei Wang
  • Lingli Chen
  • Yunyi Kong
  • Peipei Zhang
  • Zheng Liu
  • Quan Zhou
  • Yifan Chen
  • Jiajun Wang
  • Qi Bai
  • Yu Xia
  • Li Liu
  • Yu Zhu
  • Le Xu
  • Bo Dai
  • Jianming Guo
  • Yiwei WangEmail author
  • Jiejie XuEmail author
  • Weijuan ZhangEmail author
Original Article

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-9TAMs 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.

Keywords

Galectin-9+ tumor-associated macrophages Muscle-invasive bladder cancer Adjuvant chemotherapy Immune contexture 

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-9TAMs

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

Notes

Author 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.

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.

Compliance with ethical standards

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.

Supplementary material

262_2019_2429_MOESM1_ESM.pdf (487 kb)
Supplementary material 1 (PDF 486 kb)

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Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Department of Immunology, School of Basic Medical SciencesFudan UniversityShanghaiChina
  2. 2.Department of UrologyFudan University Shanghai Cancer CenterShanghaiChina
  3. 3.Department of UrologyZhongshan Hospital, Fudan UniversityShanghaiChina
  4. 4.Department of PathologyZhongshan Hospital, Fudan UniversityShanghaiChina
  5. 5.Department of PathologyFudan University Shanghai Cancer CenterShanghaiChina
  6. 6.Department of PathologyRuijin Hospital, Shanghai Jiao Tong University School of MedicineShanghaiChina
  7. 7.Department of Biochemistry and Molecular Biology, School of Basic Medical SciencesFudan UniversityShanghaiChina
  8. 8.Department of UrologyRuijin Hospital, Shanghai Jiao Tong University School of MedicineShanghaiChina
  9. 9.Department of Urology, Shanghai Ninth People’s HospitalShanghai Jiao Tong University School of MedicineShanghaiChina

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