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Circulating extracellular vesicles carrying Firmicutes reflective of the local immune status may predict clinical response to pembrolizumab in urothelial carcinoma patients

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Abstract

Bacterial flora has clinical significance for the host. The metabolic environment created by this flora influences immunotherapy in urothelial carcinoma. However, there are no reports on the clinical significance of bacterial flora in the host bloodstream. We aimed to clarify the correlation between extracellular vesicle (EV)-derived blood microflora information and tumor immunological status in urothelial carcinoma (UC) patients. Serum samples were collected from 20 healthy donors, 50 patients with localized UC, and 31 patients with metastatic UC (mUC) who had undergone pembrolizumab treatment. Bacterial DNA in EVs was extracted from each sample. Metagenomic sequencing was performed after amplification of the V1–V2 region of the bacterial 16S rRNA gene. Using the matched tumor tissue and serum samples, we revealed that the smaller amount of peripheral EVs carrying Firmicutes DNA was significantly correlated with the higher number of infiltrating T cells within tumor tissues (CD3; p = 0.015, CD4; p = 0.039, CD8; p = 0.0084) and the higher expression of activation markers on their surface (ICOS on both CD4; p = 0.0013 and CD8 T cells; p = 0.016 and 4-1BB on CD4 T cells; p = 0.016). In terms of circulating metabolic information, l-Ser and l-Pro levels, which play important roles in T cell expansion and proliferation, were significantly higher in the Firmicutes-low group (p = 0.010). All of the patients with higher Firmicutes abundance had disease progression without any clinical response (p = 0.026) and significantly inferior prognosis for pembrolizumab therapy (p = 0.035). This is the first study on the importance of peripheral bacterial EVs in cancer patients treated with cancer immunotherapy.

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The data generated in this study are available within the article and its supplementary data files.

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Funding

Japan Society for the Promotion of Science Grants-in-Aid for Scientific Research (C) (19K09709 awarded to AK); and the Suzuki Urological Foundation in Japan (awarded to AK).

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Author notes

  1. Kentaro Jingushi, Atsunari Kawashima and Takuro Saito have contributed equally to this work.

Authors and Affiliations

  1. Laboratory of Molecular and Cellular Physiology, Graduate School of Pharmaceutical Sciences, Osaka University, Suita, Osaka, Japan

    Kentaro Jingushi & Kazutake Tsujikawa

  2. Department of Urology, Graduate School of Medicine, Osaka University, Suita, Osaka, Japan

    Atsunari Kawashima, Akinaru Yamamoto, Toshihiro Uemura, Gaku Yamamichi, Koichi Okada, Eisuke Tomiyama, Yoko Koh, Makoto Matsushita, Taigo Kato, Koji Hatano, Motohide Uemura & Norio Nonomura

  3. Department of Surgery, Graduate School of Medicine, Osaka University, Suita, Osaka, Japan

    Takuro Saito

  4. Department of Clinical Research in Tumor Immunology, Graduate School of Medicine, Osaka University, Suita, Osaka, Japan

    Takuro Saito, Takayuki Kanazawa & Hisashi Wada

  5. Drug Discovery & Disease Research Laboratory, Shionogi & Co. Ltd, Toyonaka, Osaka, Japan

    Takayuki Kanazawa

  6. Research Institute for Microbial Diseases, Osaka University, Suita, Osaka, Japan

    Daisuke Motooka

  7. Reverse Translational Project, Center for Rare Disease Research, National Institute of Biomedical Innovation, Health and Nutrition (NIBIOHN), Minoh, Osaka, Japan

    Tomonori Kimura

  8. KAGAMI Inc., Ibaraki, Osaka, Japan

    Masashi Mita

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  1. Kentaro Jingushi
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Contributions

Conceptualization was done by KJ, AK, and TS, Methodology was done by KJ, AK, TK, DM, TK, MM, AY, TU, GY, KO, YK, ET, and MU; Investigation was done by KJ, AK, and TS; Visualization was done by KJ and AK; Funding acquisition was don by KJ, AK, HW, and NN; Project administration was done by AK; Supervision was done by KT, HW, and NN; Writing—original draft were done by KJ and AK; Writing—review and editing were done by KJ, AK, TS, TK, MM, KT, HW, and NN. All authors read and approved the final manuscript.

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Correspondence to Atsunari Kawashima.

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The study was approved by the ethics review board of both the Osaka University Medical Hospital (#13397–2, #14069–3).

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Jingushi, K., Kawashima, A., Saito, T. et al. Circulating extracellular vesicles carrying Firmicutes reflective of the local immune status may predict clinical response to pembrolizumab in urothelial carcinoma patients. Cancer Immunol Immunother 71, 2999–3011 (2022). https://doi.org/10.1007/s00262-022-03213-5

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