PD-1+ TIM-3+ T cells in malignant ascites predict prognosis of gastrointestinal cancer

Michitaka Nakano, Mamoru Ito, Risa Tanaka, Kyoko Yamaguchi, Hiroshi Ariyama, Kenji Mitsugi, Tomoyasu Yoshihiro, Hirofumi Ohmura, Nobuhiro Tsuruta, Fumiyasu Hanamura, Kosuke Sagara, Yuta Okumura, Kenta Nio, Kenji Tsuchihashi, Shuji Arita, Hitoshi Kusaba, Koichi Akashi, Eishi Baba, Michitaka Nakano, Mamoru Ito, Risa Tanaka, Kyoko Yamaguchi, Hiroshi Ariyama, Kenji Mitsugi, Tomoyasu Yoshihiro, Hirofumi Ohmura, Nobuhiro Tsuruta, Fumiyasu Hanamura, Kosuke Sagara, Yuta Okumura, Kenta Nio, Kenji Tsuchihashi, Shuji Arita, Hitoshi Kusaba, Koichi Akashi, Eishi Baba

Abstract

The liquid biopsy of ascites fluid could be an excellent source of tumor and microenvironment for the study of prognostic biomarkers because of its accessibility. Tumor-infiltrating lymphocytes (TILs) can predict prognosis in multiple malignancies, including the response to immune checkpoint inhibitors, a breakthrough cancer therapy. However, TILs' profiles from malignant ascites have not been extensively studied. Using flow cytometric analysis, we quantified the proportion of exhausted T cells and memory/naive/effector T-cell subsets, among the CD4+ and CD8+ T-cell populations of paired TILs and peripheral blood T cell samples (n = 22). The correlation between CD4+ and CD8+ subset profiles suggested that the combined analysis of CD4+ and CD8+ cells in malignant ascites was clinically significant. We found that cells positive for the exhaustion markers programmed cell death-1 (PD-1), and T-cell immunoglobulin and mucin domain 3 (TIM-3), and cells coexpressing PD-1 and TIM-3 abundantly exist among malignant ascites TILs. Furthermore, patients with high frequency of PD-1+ TIM-3+ cells among the CD4+ and CD8+ T-cell population showed worse clinical outcome in multivariate analysis (n = 27). We propose that exhausted ascites TILs represent a clinically significant prognostic biomarker in advanced gastrointestinal cancer and represent an important target for immune checkpoint inhibitors.

Keywords: PD-1; ascites; exhaustion; immune checkpoint; tumor-infiltrating lymphocytes.

© 2018 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

Figures

Figure 1
Figure 1
Exhaustion marker‐positive T cells are enriched among CD4+ and CD8+ ascites tumor‐infiltrating lymphocytes (TILs). A, Representative FACS plots of ascites TILs and paired peripheral blood (PB) T cells showing programmed cell death‐1 (PD‐1)+ and T‐cell immunoglobulin and mucin domain 3 (TIM‐3)+ cells within the CD4+ and CD8+ cells (case 62). B, Summarized dot plots of the frequency of exhaustion marker‐positive cells among ascites TILs and paired PB T cells (n = 22). Median values are indicated with red lines. *P < .05; ***P < .001. C, Correlation analysis of each subset, revealing similar signatures between CD4+ and CD8+ ascites TILs (n = 22)
Figure 2
Figure 2
Exhaustion marker‐positive ascites tumor‐infiltrating lymphocytes (TILs) among CD4+ and CD8+ cells increases during disease progression. A, Schema of the experimental design. Arrows indicate time of biopsy and gray lines indicate the survival times of patients. B, Representative FACS plots of 1st and 2nd biopsy samples showing the proportion of programmed cell death‐1 (PD‐1)+ and T‐cell immunoglobulin and mucin domain 3 (TIM‐3)+ TILs among CD4+ and CD8+ T cells (case 56). C, Summarized dot plots of 1st and 2nd biopsy analyses in regard to the frequency of PD‐1+ and TIM‐3+ TILs among CD4+ and CD8+ T cells. Identical patients are indicated with colored lines (cases 49, 56, 63, and 94)
Figure 3
Figure 3
Programmed cell death‐1 (PD‐1)+ T‐cell immunoglobulin and mucin domain 3 (TIM‐3)+ exhausted ascites tumor‐infiltrating lymphocytes (TILs) within the CD4+ and CD8+ cell populations predict prognosis of gastrointestinal malignancies. A, Dot plot of PD‐1+ TIM‐3+ TILs among CD4+ and CD8+ T cells. The cut‐off value was defined by the median of the 27 cases (median CD4+, 3.4%; median CD8+, 5.9%). The correlation coefficient (r) and P value is indicated. B‐D, Kaplan‐Meier curves for overall survival of the indicated patient groups, as classified by the frequency of PD‐1+ TIM‐3+ cells among CD4+ and CD8+, CD4+, and CD8+ ascites TILs. Median overall survival (days) of each patient group is shown

References

    1. Runyon BA. Care of patients with ascites. N Engl J Med. 1994;330:337‐342.
    1. Parsons SL, Lang MW, Steele RJ, et al. Malignant ascites: a 2‐year review from a teaching hospital. Eur J Surg Oncol. 1996;22:237‐239.
    1. Maecker HT, McCoy JP, Nussenblatt R, et al. Standardizing immunophenotyping for the Human Immunology Project. Nat Rev Immunol. 2012;12:191‐200.
    1. Zarour HM. Reversing T‐cell dysfunction and exhaustion in cancer. Clin Cancer Res. 2016;22:1856‐1864.
    1. Wherry EJ, Kurachi M. Molecular and cellular insights into T cell exhaustion. Nat Rev Immunol. 2015;15:486‐499.
    1. Wherry EJ. T cell exhaustion. Nat Immunol. 2011;12:492‐499.
    1. Sakuishi K, Apetoh L, Sullivan JM, Blazar BR, Kuchroo VK, Anderson AC. Targeting Tim‐3 and PD‐1 pathways to reverse T cell exhaustion and restore anti‐tumor immunity. J Exp Med. 2010;207:2187‐2194.
    1. Zhou Q, Munger ME, Veenstra RG, et al. Coexpression of Tim‐3 and PD‐1 identifies a CD8+ T‐cell exhaustion phenotype in mice with disseminated acute myelogenous leukemia. Blood. 2011;117:4501‐4510.
    1. Jin HT, Anderson AC, Tan WG, et al. Cooperation of Tim‐3 and PD‐1 in CD8 T‐cell exhaustion during chronic viral infection. Proc Natl Acad Sci U S A. 2010;107:14733‐14738.
    1. Jia Q, Yang Y, Wan Y, et al. Tumor‐infiltrating memory T‐lymphocytes for prognostic prediction in cancer patients: a meta‐analysis. Int J Clin Exp Med. 2015;8:1803‐1813.
    1. Odorizzi PM, Pauken KE, Paley MA, Sharpe A, Wherry EJ. Genetic absence of PD‐1 promotes accumulation of terminally differentiated exhausted CD8+ T cells. J Exp Med. 2015;212:1125‐1137.
    1. Granier C, Dariane C, Combe P, et al. Tim‐3 Expression on Tumor‐Infiltrating PD‐1(+)CD8(+) T Cells Correlates with Poor Clinical Outcome in Renal Cell Carcinoma. Cancer Res. 2017;77:1075‐1082.
    1. Jiang Y, Li Y, Zhu B. T‐cell exhaustion in the tumor microenvironment. Cell Death Dis. 2015;6:e1792.

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