Skip to main content

Thank you for visiting nature.com. You are using a browser version with limited support for CSS. To obtain the best experience, we recommend you use a more up to date browser (or turn off compatibility mode in Internet Explorer). In the meantime, to ensure continued support, we are displaying the site without styles and JavaScript.

  • Article
  • Published:

Molecular Diagnostics

Pretreatment tumour immune microenvironment predicts clinical response and prognosis of muscle-invasive bladder cancer in the neoadjuvant chemotherapy setting

British Journal of Cancer volume 126, pages 606–614 (2022)Cite this article

Subjects

Abstract

Background

We examined the relationship between the tumour microenvironment and the clinical efficacy of neoadjuvant chemotherapy in patients with cT2-4aN0M0 bladder cancer using multiplex fluorescence immunohistochemistry.

Methods

The study retrospectively evaluated 51 patients who underwent radical cystectomy following neoadjuvant chemotherapy for cT2-4aN0M0 muscle-invasive bladder cancer. Patients were divided into responders (<pT2) and non-responders (≥pT2). We assessed the density of each immune cell type in intratumoural and peritumoural areas in both groups via multiplex fluorescence immunohistochemical analysis.

Results

The median age was 69 years; 39 patients were male. Twelve (23.5%), 17 (33.3%), 10 (19.7%) and 12 (23.5%) patients were pT0, pT1, pT2 and ≥pT3, respectively. Responders had a significantly higher 5-year cancer-specific survival rate (96.6%) than non-responders (48.4%; p = 0.0018). CD8+ T cell (p = 0.0056) and CD204+ cell (p = 0.0394) densities were significantly higher in the intratumoural area in non-responders than in responders. Patients with higher CD204+ cell densities in cancerous areas had worse prognosis.

Conclusions

This comprehensive analysis of the immune microenvironment of a muscle-invasive bladder cancer specimen revealed that preexisting tumour-infiltrating proliferating CD8+ T cells and CD204+ cells are indicators of the response to neoadjuvant chemotherapy and that CD204+ cells can be considered an unfavourable prognostic factor in these patients.

This is a preview of subscription content, access via your institution

Access options

Buy this article

  • Purchase on Springer Link
  • Instant access to full article PDF
Buy now

Prices may be subject to local taxes which are calculated during checkout

Fig. 1: Assessment of tumor immune microenvironment.
Fig. 2: Correlation of intratumoral immune cells in pre-NAC tissues.
Fig. 3: Changes in intratumoral immune cells pre- and post-NAC treatment.
Fig. 4: Assessment of intratumoral CD8+ T cell function.
Fig. 5: Progression-free survival and cancer-specific survival in patients with high and low infiltration of CD8+ T cell and CD204+ cell.

Similar content being viewed by others

Data availability

All data supporting the results are presented with results and in the figures.

References

  1. Gore JL, Litwin MS, Lai J, Yano EM, Madison R, Setodji C., et al. Urologic Diseases in America Project. Use of radical cystectomy for patients with invasive bladder cancer. J Natl Cancer Inst. 2010;102:802–11.

    Article  PubMed  PubMed Central  Google Scholar 

  2. Witjes JA, Bruins HM, Cathomas R, Compérat EM, Cowan NC, Gakis G, et al. European Association of Urology Guidelines on Muscle-invasive and Metastatic Bladder Cancer: summary of the 2020 guidelines. Eur Urol. 2021;79:82–104.

    Article  CAS  PubMed  Google Scholar 

  3. Chang SS, Bochner BH, Chou R, Dreicer R, Kamat AM, Lerner SP, et al. Treatment of non-metastatic muscle-invasive bladder cancer: AUA/ASCO/ASTRO/SUO Guideline. J Urol. 2017;198:552–9.

    Article  PubMed  PubMed Central  Google Scholar 

  4. Flaig TW, Spiess PE, Agarwal N, Bangs R, Boorjian SA, Buyyounouski MK, et al. NCCN Guidelines Insights: bladder cancer, version 5.2018. J Natl Compr Cancer Netw. 2018;16:1041–53.

    Article  Google Scholar 

  5. Peyton CC, Tang D, Reich RR, Azizi M, Chipollini J, Pow-Sang JM, et al. Downstaging and survival outcomes associated with neoadjuvant chemotherapy regimens among patients treated with cystectomy for muscle-invasive bladder cancer. JAMA Oncol. 2018;4:1535–42.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Pfister C, Gravis G, Fléchon A, Soulié M, Guy L, Laguerre B, et al. VESPER Trial Investigators. Randomized phase III trial of dose-dense methotrexate, vinblastine, doxorubicin, and cisplatin, or gemcitabine and cisplatin as perioperative chemotherapy for patients with muscle-invasive bladder cancer. Analysis of the GETUG/AFU V05 VESPER Trial secondary endpoints: chemotherapy toxicity and pathological responses. Eur Urol. 2021;79:214–21.

    Article  CAS  PubMed  Google Scholar 

  7. Anan G, Hatakeyama S, Fujita N, Iwamura H, Tanaka T, Yamamoto H, et al. Trends in neoadjuvant chemotherapy use and oncological outcomes for muscle-invasive bladder cancer in Japan: a multicenter study. Oncotarget. 2017;8:86130–42.

    Article  PubMed  PubMed Central  Google Scholar 

  8. Canter D, Viterbo R, Kutikov A, Wong YN, Plimack E, Zhu F, et al. Baseline renal function status limits patient eligibility to receive perioperative chemotherapy for invasive bladder cancer and is minimally affected by radical cystectomy. Urology. 2011;77:160–5.

    Article  PubMed  Google Scholar 

  9. Koie T, Ohyama C, Hashimoto Y, Hatakeyama S, Yamamoto H, Yoneyama T, et al. Efficacies and safety of neoadjuvant gemcitabine plus carboplatin followed by immediate cystectomy in patients with muscle-invasive bladder cancer, including those unfit for cisplatin: a prospective single-arm study. Int J Clin Oncol. 2013;18:724–30.

    Article  CAS  PubMed  Google Scholar 

  10. Hanahan D, Coussens LM. Accessories to the crime: functions of cells recruited to the tumor microenvironment. Cancer Cell. 2012 ;21:309–22.

    Article  CAS  PubMed  Google Scholar 

  11. Chen DS, Mellman I. Oncology meets immunology: the cancer-immunity cycle. Immunity. 2013;39:1–10.

    Article  PubMed  Google Scholar 

  12. Rosenberg JE, Hoffman-Censits J, Powles T, van der Heijden MS, Balar AV, Necchi A, et al. Atezolizumab in patients with locally advanced and metastatic urothelial carcinoma who have progressed following treatment with platinum-based chemotherapy: a single-arm, multicentre, phase 2 trial. Lancet. 2016;387:1909–20.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  13. Zeng D, Li M, Zhou R, Zhang J, Sun H, Shi M, et al. Tumor microenvironment characterization in gastric cancer identifies prognostic and immunotherapeutically relevant gene signatures. Cancer Immunol Res. 2019;7:737–50.

    Article  CAS  PubMed  Google Scholar 

  14. Jiang Y, Zhang Q, Hu Y, Li T, Yu J, Zhao L, et al. ImmunoScore Signature: a prognostic and predictive tool in gastric cancer. Ann Surg. 2018;267:504–13.

    Article  PubMed  Google Scholar 

  15. Nishino M, Ramaiya NH, Hatabu H, Hodi FS. Monitoring immunecheckpoint blockade: response evaluation and biomarker development. Nat Rev Clin Oncol. 2017;14:655–68.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. Jiang Y, Zhang Q, Hu Y, Li T, Yu J, Zhao L, et al. ImmunoScore Signature: a prognostic and predictive tool in gastric cancer. Ann Surg. 2018;267:504–13.

    Article  PubMed  Google Scholar 

  17. Hendry S, Salgado R, Gevaert T, Russell PA, John T, Thapa B, et al. Assessing tumor-infiltrating lymphocytes in solid tumors: a practical review for pathologists and proposal for a standardized method from the International Immuno-Oncology Biomarkers Working Group: Part 2: TILs in melanoma, gastrointestinal tract carcinomas, non-small cell lung carcinoma and mesothelioma, endometrial and ovarian carcinomas, squamous cell carcinoma of the head and neck, genitourinary carcinomas, and primary brain tumors. Adv Anat Pathol. 2017;24:311–35.

    Article  PubMed  PubMed Central  Google Scholar 

  18. Wang B, Pan W, Yang M, Yang W, He W, Chen X, et al. Programmed death ligand-1 is associated with tumor infiltrating lymphocytes and poorer survival in urothelial cell carcinoma of the bladder. Cancer Sci. 2019;110:489–98.

    Article  CAS  PubMed  Google Scholar 

  19. Cassetta L, Pollard JW. Targeting macrophages: therapeutic approaches in cancer. Nat Rev Drug Discov. 2018;17:887–904.

    Article  CAS  PubMed  Google Scholar 

  20. DeNardo DG, Ruffell B. Macrophages as regulators of tumour immunity and immunotherapy. Nat Rev Immunol. 2019;2019:6.

    Google Scholar 

  21. Yamamoto K, Makino T, Sato E, Noma T, Urakawa S, Takeoka T, et al. Tumor-infiltrating M2 macrophage in pretreatment biopsy sample predicts response to chemotherapy and survival in esophageal cancer. Cancer Sci. 2020;111:1103–12.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Stack EC, Wang C, Roman KA, Hoyt CC. Multiplexed immunohistochemistry, imaging, and quantitation: a review, with an assessment of Tyramide signal amplification, multispectral imaging and multiplex analysis. Methods. 2014;70:46–58.

    Article  CAS  PubMed  Google Scholar 

  23. Igari F, Sato E, Horimoto Y, Takahashi Y, Isomura T, Arakawa A, et al. Diagnostic significance of intratumoral CD8+ tumor-infiltrating lymphocytes in medullary carcinoma. Hum Pathol. 2017;70:129–38.

    Article  CAS  PubMed  Google Scholar 

  24. Imaizumi K, Suzuki T, Kojima M, Shimomura M, Sakuyama N, Tsukada Y, et al. Ki67 expression and localization of T cells after neoadjuvant therapies as reliable predictive markers in rectal cancer. Cancer Sci. 2020;111:23–35.

    Article  CAS  PubMed  Google Scholar 

  25. Salgado R, Denkert C, Demaria S, Sirtaine N, Klauschen F, Pruneri G, et al. The evaluation of tumor-infiltrating lymphocytes (TILs) in breast cancer: recommendations by an International TILs Working Group 2014. Ann Oncol. 2015;26:259–71.

    Article  CAS  PubMed  Google Scholar 

  26. Hayashi T, Kohsaka S, Takamochi K, Kishikawa S, Ikarashi D, Sano K, et al. Histological characteristics of lung adenocarcinoma with uncommon actionable alterations: special emphasis on MET exon 14 skipping alterations. Histopathology. 2021;78:987–99.

    Article  PubMed  Google Scholar 

  27. Ikarashi D, Kitano S, Ishida K, Nakatsura T, Shimodate H, Tsuyukubo T, et al. Complete pathological response to neoadjuvant pembrolizumab in a patient with chemoresistant upper urinary tract urothelial carcinoma: a case report. Front Oncol. 2020;10:564714.

    Article  PubMed  PubMed Central  Google Scholar 

  28. Zhang QW, Liu L, Gong CY, Shi HS, Zeng YH, Wang XZ, et al. Prognostic significance of tumor-associated macrophages in solid tumor: a meta-analysis of the literature. PLoS ONE. 2012;7:e50946.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Takayama H, Nishimura K, Tsujimura A, Nakai Y, Nakayama M, Aozasa K, et al. Increased infiltration of tumor associated macrophages is associated with poor prognosis of bladder carcinoma in situ after intravesical bacillus Calmette-Guerin instillation. J Urol. 2009;181:1894–900.

    Article  CAS  PubMed  Google Scholar 

  30. Tervahartiala M, Taimen P, Mirtti T, Koskinen I, Ecke T, Jalkanen S, et al. Immunological tumor status may predict response to neoadjuvant chemotherapy and outcome after radical cystectomy in bladder cancer. Sci Rep. 2017;7:12682.

    Article  PubMed  PubMed Central  Google Scholar 

  31. Wang X, Ni S, Chen Q, Ma L, Jiao Z, Wang C, et al. Bladder cancer cells induce immunosuppression of T cells by supporting PD-L1 expression in tumour macrophages partially through interleukin 10. Cell Biol Int. 2017;41:177–86.

    Article  CAS  PubMed  Google Scholar 

  32. Sharifi L, Nowroozi MR, Amini E, Arami MK, Ayati M, Mohsenzadegan M. A review on the role of M2 macrophages in bladder cancer; pathophysiology and targeting. Int Immunopharmacol. 2019;76:105880.

    Article  CAS  PubMed  Google Scholar 

  33. Aljabery F, Olsson H, Gimm O, Jahnson S, Shabo I. M2-macrophage infiltration and macrophage traits of tumor cells in urinary bladder cancer. Urol Oncol. 2018;36:159.e19–159.e26.

    Article  CAS  Google Scholar 

  34. Sharifi L, Nowroozi MR, Amini E, Arami MK, Ayati M, Mohsenzadegan M. A review on the role of M2 macrophages in bladder cancer; pathophysiology and targeting. Int Immunopharmacol. 2019;76:105880.

    Article  CAS  PubMed  Google Scholar 

  35. DeNardo DG, Ruffell B. Macrophages as regulators of tumour immunity and immunotherapy. Nat Rev Immunol. 2019;19:369–82.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  36. Yang L, Li A, Liu F, Zhao Q, Ji S, Zhu W, et al. Immune profiling reveals molecular classification and characteristic in urothelial bladder cancer. Front Cell Dev Biol. 2021;9:596484.

    Article  PubMed  PubMed Central  Google Scholar 

  37. Choi W, Porten S, Kim S, Willis D, Plimack ER, Hoffman-Censits J, et al. Identification of distinct basal and luminal subtypes of muscle-invasive bladder cancer with different sensitivities to frontline chemotherapy. Cancer Cell. 2014;25:152–65.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  38. Saito T, Nishikawa H, Wada H, Nagano Y, Sugiyama D, Atarashi K, et al. Two FOXP3(+)CD4(+) T cell subpopulations distinctly control the prognosis of colorectal cancers. Nat Med. 2016;22:679–84.

    Article  CAS  PubMed  Google Scholar 

  39. Takeuchi Y, Nishikawa H. Roles of regulatory T cells in cancer immunity. Int Immunol. 2016;28:401–9.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  40. Tan WCC, Nerurkar SN, Cai HY, Ng HHM, Wu D, Wee YTF, et al. Overview of multiplex immunohistochemistry/immunofluorescence techniques in the era of cancer immunotherapy. Cancer Commun. 2020;40:135–53.

    Article  Google Scholar 

  41. Necchi A, Anichini A, Raggi D, Briganti A, Massa S, Lucianò R, et al. Pembrolizumab as neoadjuvant therapy before radical cystectomy in Patients With Muscle-Invasive Urothelial Bladder Carcinoma (PURE-01): an open-label, single-arm, phase II study. J Clin Oncol. 2018;36:3353–60.

    Article  CAS  PubMed  Google Scholar 

  42. Topalian SL, Taube JM, Pardoll DM. Neoadjuvant checkpoint blockade for cancer immunotherapy. Science. 2020;367:eaax0182.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  43. Wang L, Sfakianos JP, Beaumont KG, Akturk G, Horowitz A, Sebra RP, et al. Myeloid cell-associated resistance to PD-1/PD-L1 blockade in urothelial cancer revealed through bulk and single-cell RNA sequencing. Clin Cancer Res. 2021;27:4287–300.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Nywening TM, Wang-Gillam A, Sanford DE, Belt BA, Panni RZ, Cusworth BM, et al. Targeting tumour-associated macrophages with CCR2 inhibition in combination with FOLFIRINOX in patients with borderline resectable and locally advanced pancreatic cancer: a single-centre, open-label, dose-finding, non-randomised, phase 1b trial. Lancet Oncol. 2016;17:651–62.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

Download references

Acknowledgements

The authors would like to thank all patients and their families. They also would like to thank Enago (www.enago.jp) for the English language review.

Author information

Authors and Affiliations

  1. Department of Urology, Iwate Medical University School of Medicine, Iwate, 028-3695, Japan

    Daiki Ikarashi, Takashi Tsuyukubo, Shigekatsu Maekawa, Renpei Kato, Yoichiro Kato & Wataru Obara

  2. Division of Cancer Immunotherapy Development, Advanced Medical Development Center, The Cancer Institute Hospital of Japanese Foundation for Cancer Research, Tokyo, 135-8550, Japan

    Daiki Ikarashi, Shigehisa Kitano, Makiko Yamashita & Hidenori Mizugaki

  3. Division of Cancer Immunotherapy, Exploratory Oncology Research and Clinical Trial Center, National Cancer Center, Chiba, 277-0872, Japan

    Daiki Ikarashi, Kazumasa Takenouchi, Takayuki Nakayama & Tetsuya Nakatsura

  4. Department of Human Pathology, Juntendo University School of Medicine, Tokyo, 113-0033, Japan

    Hiroko Onagi & Asumi Sakaguchi

  5. Department of Pathology, Iwate Medical University School of Medicine, Iwate, 028-3695, Japan

    Tamotsu Sugai

Authors
  1. Daiki Ikarashi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Shigehisa Kitano
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Takashi Tsuyukubo
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kazumasa Takenouchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Takayuki Nakayama
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hiroko Onagi
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Asumi Sakaguchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Makiko Yamashita
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Hidenori Mizugaki
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Shigekatsu Maekawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Renpei Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Yoichiro Kato
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Tamotsu Sugai
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Tetsuya Nakatsura
    View author publications

    You can also search for this author in PubMed Google Scholar

  15. Wataru Obara
    View author publications

    You can also search for this author in PubMed Google Scholar

Contributions

Study conception and design: DI, SK and WO; data acquisition and analysis: DI, TT, KT, MY, HM, SM, RK and YK; drafting the manuscript and figures: DI, HO and AS; reviewing the manuscript: SK, TN, TS, TT and WO. All authors have read and approved the final draft for submission.

Corresponding author

Correspondence to Shigehisa Kitano.

Ethics declarations

Competing interests

SK reports personal fees from Astra Zeneca, grants and personal fees from Chugai, personal fees from Pfizer, grants and personal fees from Boehringer Ingelheim, personal fees from Taiho, personal fees from Novartis, grants and personal fees from Daiichi-Sankyo, personal fees from MSD, personal fees from Sumitomo Dainippon Pharma, grants and personal fees from Eisai, grants from Astellas, grants from Gilead Sciences, grants and personal fees from Ono Pharmaceutical Co., Ltd, personal fees from Bristol-Myers Squibb, grants and personal fees from REGENERON, personal fees from AYUMI Pharmaceutical Corporation, personal fees from Rakuten Medical, grants from PACT Pharma, grants from Takara Bio Inc., personal fees from GSK, personal fees from ImmuniT Research Inc., grants and personal fees from Ono Pharmaceutical Co., Ltd, personal fees from PMDA (Pharmaceuticals and Medical Devices Agency), grants from AMED (Japan Agency for Medical Research and Development) and grants from JSPS (Japan Society for the Promotion of Science), outside the submitted work. The remaining authors declare no competing interests.

Ethics approval and consent to participate

The study was conducted in accordance with the principles of the Declaration of Helsinki. The human ethics board of each institution approved this study and written informed consent was obtained from all patients prior to enrolment (Iwate Medical University; Protocol No. 2019-083, National Cancer Center Hospital East; Protocol No. 2019-194).

Additional information

Publisher’s note Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Supplementary information

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Ikarashi, D., Kitano, S., Tsuyukubo, T. et al. Pretreatment tumour immune microenvironment predicts clinical response and prognosis of muscle-invasive bladder cancer in the neoadjuvant chemotherapy setting. Br J Cancer 126, 606–614 (2022). https://doi.org/10.1038/s41416-021-01628-y

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1038/s41416-021-01628-y

This article is cited by

Search

Advanced search

Quick links