LAG-3 expression on tumor-infiltrating T cells in soft tissue sarcoma correlates with poor survival
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摘要
Objective: To elucidate the role and prognostic significance of lymphocyte activation-gene-3(LAG-3) in soft tissue sarcoma(STS).Methods: The expression of LAG-3 in patient and matched normal blood samples was analyzed by flow cytometry. The localization and prognostic values of LAG-3~+ cells in 163 STS patients were analyzed by immunohistochemistry. In addition, the expression of tumor-infiltrating CD3~+ T, CD4~+ T, and CD8~+ T cells and their role in the prognosis of STS were evaluated by immunohistochemistry. The effect of LAG-3 blockade was evaluated in an immunocompetent MCA205 fibrosarcoma mouse model.Results: Peripheral CD8~+ and CD4~+ T cells from STS patients expressed higher levels of LAG-3 than those from healthy donors.LAG-3 expression in STS was significantly associated with a poor clinical outcome(P = 0.038) and was correlated with high pathological grade(P < 0.001), advanced tumor stage(P = 0.016). Additionally, LAG-3 expression was highly correlated with CD8~+ T-cell infiltration(r = 0.7034, P < 0.001). LAG-3 was expressed in murine tumor-infiltrating lymphocytes, and its blockade decreased tumor growth and enhanced secretion of interferon-gamma by CD8~+ and CD4~+ T cells.Conclusions: LAG-3 blockade may be a promising strategy to improve the effects of targeted therapy in STS.
Objective: To elucidate the role and prognostic significance of lymphocyte activation-gene-3(LAG-3) in soft tissue sarcoma(STS).Methods: The expression of LAG-3 in patient and matched normal blood samples was analyzed by flow cytometry. The localization and prognostic values of LAG-3~+ cells in 163 STS patients were analyzed by immunohistochemistry. In addition, the expression of tumor-infiltrating CD3~+ T, CD4~+ T, and CD8~+ T cells and their role in the prognosis of STS were evaluated by immunohistochemistry. The effect of LAG-3 blockade was evaluated in an immunocompetent MCA205 fibrosarcoma mouse model.Results: Peripheral CD8~+ and CD4~+ T cells from STS patients expressed higher levels of LAG-3 than those from healthy donors.LAG-3 expression in STS was significantly associated with a poor clinical outcome(P = 0.038) and was correlated with high pathological grade(P < 0.001), advanced tumor stage(P = 0.016). Additionally, LAG-3 expression was highly correlated with CD8~+ T-cell infiltration(r = 0.7034, P < 0.001). LAG-3 was expressed in murine tumor-infiltrating lymphocytes, and its blockade decreased tumor growth and enhanced secretion of interferon-gamma by CD8~+ and CD4~+ T cells.Conclusions: LAG-3 blockade may be a promising strategy to improve the effects of targeted therapy in STS.
Objective: To elucidate the role and prognostic significance of lymphocyte activation-gene-3(LAG-3) in soft tissue sarcoma(STS).Methods: The expression of LAG-3 in patient and matched normal blood samples was analyzed by flow cytometry. The localization and prognostic values of LAG-3~+ cells in 163 STS patients were analyzed by immunohistochemistry. In addition, the expression of tumor-infiltrating CD3~+ T, CD4~+ T, and CD8~+ T cells and their role in the prognosis of STS were evaluated by immunohistochemistry. The effect of LAG-3 blockade was evaluated in an immunocompetent MCA205 fibrosarcoma mouse model.Results: Peripheral CD8~+ and CD4~+ T cells from STS patients expressed higher levels of LAG-3 than those from healthy donors.LAG-3 expression in STS was significantly associated with a poor clinical outcome(P = 0.038) and was correlated with high pathological grade(P < 0.001), advanced tumor stage(P = 0.016). Additionally, LAG-3 expression was highly correlated with CD8~+ T-cell infiltration(r = 0.7034, P < 0.001). LAG-3 was expressed in murine tumor-infiltrating lymphocytes, and its blockade decreased tumor growth and enhanced secretion of interferon-gamma by CD8~+ and CD4~+ T cells.Conclusions: LAG-3 blockade may be a promising strategy to improve the effects of targeted therapy in STS.
引文
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5. Coley II WB.Contribution to the Knowledge of Sarcoma.Ann Surg.1891;14:199-220.
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8. Nowicki TS,Akiyama R,Huang RR,Shintaku IP,Wang XY,Tumeh PC,et al.Infiltration of CD8T cells and expression of PD-1and PD-Ll1in synovial sarcoma.Cancer Immunol Res.2017;5:118-26.
9. Baksh K,Weber J.Immune checkpoint protein inhibition for cancer:preclinical justification for CTLA-4 and PD-1 blockade and new combinations.Semin Oncol.2015;42:363-77.
10. Keenan BP,Jaffee EM,Armstrong TD.Tumor immunology:multidisciplinary science driving basic and clinical advances.Cancer Immunol Res.2013;1:16-23.
11. Robert C,Thomas L,Bondarenko I,O'Day S,Weber J,Garbe C,et al.Ipilimumab plus dacarbazine for previously untreated metastatic melanoma.N Engl J Med.2011;364:2517-26.
12. Couzin-Frankel J.Cancer immunotherapy.Science.2013;342:1432-33.
13. Brahmer JR,Tykodi SS,Chow LQ,Hwu WJ,Topalian SL,Hwu P,et al.Safety and activity of anti-PD-Ll antibody in patients with advanced cancer.N Engl J Med.2012;366:2455-65.
14. Hodi FS,O'Day SJ,McDermott DF,Weber RW,Sosman JA,Haanen JB,et al.Improved survival with ipilimumab in patients with metastatic melanoma.N Engl J Med.2010;363:711-23.
15. Tawbi HA,Burgess M,Bolejack V,Van Tine BA,Schuetze SM,Hu J,et al.Pembrolizumab in advanced soft-tissue sarcoma and bone sarcoma(SARC028):a multicentre,two-cohort,single-arm,openlabel,phase 2 trial.Lancet Oncol.2017;18:1493-501.
16. Butler NS,Moebius J,Pewe LL,Traore B,Doumbo OK,Tygrett LT,et al.Therapeutic blockade of PD-L1 and LAG-3 rapidly clears established blood-stage Plasmodium infection.Nat Immunol.2011;13:188-95.
17. Matsuzaki J,Gnjatic S,Mhawech-Fauceglia P,Beck A,Miller A,Tsuji T,et al.Tumor-infiltrating NY-ESO-1-specific CD8+T cells are negatively regulated by LAG-3 and PD-1 in human ovarian cancer.Proc Natl Acad Sci USA.2010;107:7875-80.
18. Huard B,Prigent P,Tournier M,Bruniquel D,Triebel F.CD4/maj or histocompatibility complex classⅡinteraction analyzec with CD4-and lymphocyte activation gene-3(LAG-3)-Ig fusion proteins.Eur J Immunol.1995;25:2718-21.
19. Xu F,Liu J,Liu D,Liu B,Wane M,Hu ZY,et al.LSECtin expressedon melanoma cells promotes tumor progression by inhibiting antitumor T-cell responses.Cancer Res.2014;74:3418-28.
20. Kotaskova J,Tichy B,Trbusek M,Francova HS,Kabathova J,Malcikova J,et al.High expression of Lymphocyte-Activation gene3(LA G3)in chronic lymphocytic leukemia cells is associated with unmutated Immunoglobulin variable heavy Chain region(IGHV)gene and reduced treatment-free survival.J Mol Diagn.2010;12:328-34.
21. Li FJ,Zhang Y,Jin GX,Yao L,Wu DQ.Expression of LAG-3 is coincident with the impaired effector function of HBV-specific CD8+T cell in HCC patients.Immunol Lett.2013;150:116-22.
22. Takaya S,Saito H,Ikeguchi M.Upregulation of immune checkpoint molecules,PD-1 and LAG-3,on CD4+and CD8+T cells after gastric cancer surgery.Yonago Acta Med.2015;58:39-44.
23. Gros A,Robbins PF,Yao X,Li YF,Turcotte S,Tran E,et al.PD-1identifies the patient-specific CD8+tumor-reactive repertoire infiltrating human tumors.J Clin Invest.2014;124:2246-59.
24. Woo SR,Turnis ME,Goldberg MV,Bankoti J,Selby M,Nirschl CJ,et al.Immune inhibitory molecules LAG-3 and PD-1 synergistically regulate T cell function to promote tumoral immune escape.Cancer Res.2012;72:917-27.
25. Toro JR,Travis LB,Wu HJ,Zhu KM,Fletcher CDM,Devesa SS.Incidence patterns of soft tissue sarcomas,regardless of primary site,in the surveillance,epidemiology and end results program,1978-2001:an analysis of 26,758 cases.Int J Cancer.2006;119:2922-30.
26. Yu GT,Bu LL,Huang CF,Zhang WF,Chen WJ,Gutkind JS,et al.PD-1 blockade attenuates immunosuppressive myeloid cells due to inhibition of CD47/SIRPαaxis in HPV negative head and neck squamous cell carcinoma.Oncotarget.2015;6:42067-80.
27. D'Angelo SP,Shoushtari AN,Agaram NP,Kuk D,Qin LX,Carvajal RD,et al.Prevalence of tumor-infiltrating lymphocytes and PD-L1expression in the soft tissue sarcoma microenvironment.Human Pathol.2015;46:357-65.
28. Hou JY,Yu Z,Xiang RY,Li CQ,Wang L,Chen SF,et al.Correlation between infiltration of FOXP3+regulatory T cells and expression of B7-H1 in the tumor tissues of gastric cancer.Exp Mol Pathol.2014;96:284-91.
29. Deng WW,Mao L,Yu GT,Bu LL,Ma SR,Liu B,et al.LAG-3confers poor prognosis and its blockade reshapes antitumor response in head and neck squamous cell carcinoma.Oncoimmunology.2016;5:e1239005.
30. Chen DS,Mellman I.Oncology meets immunology:the cancerimmunity cycle.Immunity.2013;39:1-10.
31. Topalian SL,Drake CG,Pardoll DM.Immune checkpoint blockade:a common denominator approach to cancer therapy.Cancer Cell.2015;27:450-61.
32. Liu JF,Wu L,Yang LL,Deng WW,Mao L,Wu H,et al.Blockade ofTIM3 relieves immunosuppression through reducing regulatory T cells in head and neck cancer.J Exp Clin Cancer Res.2018;37:44.
33. Fridman WH,Pages F,Sautes-Fridman C,Galon J.The immune contexture in human tumours:impact on clinical outcome.Nat Rev Cancer.2012;12:298-306.
34. Muris JJ,Meijer CJ,Cillessen SA,Vos W,Kummer JA,Bladergroen BA,et al.Prognostic significance of activated cytotoxic Tlymphocytes in primary nodal diffuse large B-cell lymphomas.Leukemia.2004;18:589-96.
35. Scott DW,Chan FC,Hong FX,Rogic S,Tan KL,Meissner B,et al.Gene expression-based model using formalin-fixed paraffinembedded biopsies predicts overall survival in advanced-stage classical Hodgkin lymphoma.J Clin Oncol.2013;31:692-700.
36. Nakano O,Sato M,Naito Y,Suzuki K,Orikasa S,Aizawa M,et al.Proliferative activity of intratumoral CD8+T-lymphocytes as a prognostic factor in human renal cell carcinoma:clinicopathologic demonstration of antitumor immunity.Cancer Res.2001;61:5132-36.
37. Sorbye SW,Kilvaer T,Valkov A,Donnem T,Smeland E,Al-Shibli K,et al.Prognostic impact of lymphocytes in soft tissue sarcomas.PLoS One.2011;6:e14611.
38. Grosso JF,Kelleher CC,Harris TJ,Maris CH,Hipkiss EL,De Marzo A,et al.LAG-3 regulates CD8+T cell accumulation and effector function in murine self-and tumor-tolerance systems.J Clin Invest.2007;117:3383-92.
39. Hannier S,Triebel F.The MHC class II ligand lymphocyte activation gene-3 is co-distributed with CD8 and CD3-TCR molecules after their engagement by mAb or peptide-MHC class I complexes.Int Immunol.1999;11:1745-52.
40. Toulmonde M,Penel N,Adam J,Chevreau C,Blay JY,Le Cesne A.et al.Use of PD-1 targeting,macrophage infiltration,and IDO pathway activation in sarcomas:a phase 2 clinical trial.JAMA Oncol.2018;4:93-7.
41. D'Angelo SP,Shoushtari AN,Keohan ML,Dickson MA,Gounder MM,Chi P,et al.Combined KIT and CTLA-4 Blockade in Patients with Refractory GIST and other advanced sarcomas:a phase ib study of dasatinib plus Ipilimumab.Clin Cancer Res.2017;23:2972-80.
42. Triebel F,Jitsukawa S,Baixeras E,Roman-Roman S,Genevee C,Viegas-Pequignot E,et al.LAG-3,a novel lymphocyte activation gene closely related to CD4.J Exp Med.1990;171:1393-405.
2. Sleijfer S,Ouali M,van Glabbeke M,Krarup-Hansen A,Rodenhuis S,Le Cesne A,et al.Prognostic and predictive factors for outcome to first-line ifosfamide-containing chemotherapy for adult patients with advanced soft tissue sarcomas:an exploratory,retrospective analysis on large series from the European Organization for Research and Treatment of Cancer-Soft Tissue and Bone Sarcoma Group(EORTC-STBSG).Eur J Cancer.2010;46:72-83.
3. Daigeler A,Zmarsly I,Hirsch T,Goertz O,Steinau HU,LehnhardtM,et al.Long-term outcome after local recurrence of soft tissue sarcoma:a retrospective analysis of factors predictive of survival in135 patients with locally recurrent soft tissue sarcoma.Br J Cancer.2014;110:1456-64.
4. Weitz J,Antonescu CR,Brennan MF. Localized extremity soft tissue sarcoma:improved knowledge with unchanged survival over time.J Clin Oncol.2003;21:2719-25.
5. Coley II WB.Contribution to the Knowledge of Sarcoma.Ann Surg.1891;14:199-220.
6. Johnston BJ,Novales ET.Clinical effect of Coley's toxin.II.A seven-year study.Cancer Chemother Rep.1962;21:43-68.
7. Robbins PF,Morgan RA,Feldman SA,Yang JC,Sherry RM,Dudley ME,et al.Tumor regression in patients with metastatic synovial cell sarcoma and melanoma using genetically engineered lymphocytes reactive with NY-ESO-1.J Clin Oncol.2011;29:917-24.
8. Nowicki TS,Akiyama R,Huang RR,Shintaku IP,Wang XY,Tumeh PC,et al.Infiltration of CD8T cells and expression of PD-1and PD-Ll1in synovial sarcoma.Cancer Immunol Res.2017;5:118-26.
9. Baksh K,Weber J.Immune checkpoint protein inhibition for cancer:preclinical justification for CTLA-4 and PD-1 blockade and new combinations.Semin Oncol.2015;42:363-77.
10. Keenan BP,Jaffee EM,Armstrong TD.Tumor immunology:multidisciplinary science driving basic and clinical advances.Cancer Immunol Res.2013;1:16-23.
11. Robert C,Thomas L,Bondarenko I,O'Day S,Weber J,Garbe C,et al.Ipilimumab plus dacarbazine for previously untreated metastatic melanoma.N Engl J Med.2011;364:2517-26.
12. Couzin-Frankel J.Cancer immunotherapy.Science.2013;342:1432-33.
13. Brahmer JR,Tykodi SS,Chow LQ,Hwu WJ,Topalian SL,Hwu P,et al.Safety and activity of anti-PD-Ll antibody in patients with advanced cancer.N Engl J Med.2012;366:2455-65.
14. Hodi FS,O'Day SJ,McDermott DF,Weber RW,Sosman JA,Haanen JB,et al.Improved survival with ipilimumab in patients with metastatic melanoma.N Engl J Med.2010;363:711-23.
15. Tawbi HA,Burgess M,Bolejack V,Van Tine BA,Schuetze SM,Hu J,et al.Pembrolizumab in advanced soft-tissue sarcoma and bone sarcoma(SARC028):a multicentre,two-cohort,single-arm,openlabel,phase 2 trial.Lancet Oncol.2017;18:1493-501.
16. Butler NS,Moebius J,Pewe LL,Traore B,Doumbo OK,Tygrett LT,et al.Therapeutic blockade of PD-L1 and LAG-3 rapidly clears established blood-stage Plasmodium infection.Nat Immunol.2011;13:188-95.
17. Matsuzaki J,Gnjatic S,Mhawech-Fauceglia P,Beck A,Miller A,Tsuji T,et al.Tumor-infiltrating NY-ESO-1-specific CD8+T cells are negatively regulated by LAG-3 and PD-1 in human ovarian cancer.Proc Natl Acad Sci USA.2010;107:7875-80.
18. Huard B,Prigent P,Tournier M,Bruniquel D,Triebel F.CD4/maj or histocompatibility complex classⅡinteraction analyzec with CD4-and lymphocyte activation gene-3(LAG-3)-Ig fusion proteins.Eur J Immunol.1995;25:2718-21.
19. Xu F,Liu J,Liu D,Liu B,Wane M,Hu ZY,et al.LSECtin expressedon melanoma cells promotes tumor progression by inhibiting antitumor T-cell responses.Cancer Res.2014;74:3418-28.
20. Kotaskova J,Tichy B,Trbusek M,Francova HS,Kabathova J,Malcikova J,et al.High expression of Lymphocyte-Activation gene3(LA G3)in chronic lymphocytic leukemia cells is associated with unmutated Immunoglobulin variable heavy Chain region(IGHV)gene and reduced treatment-free survival.J Mol Diagn.2010;12:328-34.
21. Li FJ,Zhang Y,Jin GX,Yao L,Wu DQ.Expression of LAG-3 is coincident with the impaired effector function of HBV-specific CD8+T cell in HCC patients.Immunol Lett.2013;150:116-22.
22. Takaya S,Saito H,Ikeguchi M.Upregulation of immune checkpoint molecules,PD-1 and LAG-3,on CD4+and CD8+T cells after gastric cancer surgery.Yonago Acta Med.2015;58:39-44.
23. Gros A,Robbins PF,Yao X,Li YF,Turcotte S,Tran E,et al.PD-1identifies the patient-specific CD8+tumor-reactive repertoire infiltrating human tumors.J Clin Invest.2014;124:2246-59.
24. Woo SR,Turnis ME,Goldberg MV,Bankoti J,Selby M,Nirschl CJ,et al.Immune inhibitory molecules LAG-3 and PD-1 synergistically regulate T cell function to promote tumoral immune escape.Cancer Res.2012;72:917-27.
25. Toro JR,Travis LB,Wu HJ,Zhu KM,Fletcher CDM,Devesa SS.Incidence patterns of soft tissue sarcomas,regardless of primary site,in the surveillance,epidemiology and end results program,1978-2001:an analysis of 26,758 cases.Int J Cancer.2006;119:2922-30.
26. Yu GT,Bu LL,Huang CF,Zhang WF,Chen WJ,Gutkind JS,et al.PD-1 blockade attenuates immunosuppressive myeloid cells due to inhibition of CD47/SIRPαaxis in HPV negative head and neck squamous cell carcinoma.Oncotarget.2015;6:42067-80.
27. D'Angelo SP,Shoushtari AN,Agaram NP,Kuk D,Qin LX,Carvajal RD,et al.Prevalence of tumor-infiltrating lymphocytes and PD-L1expression in the soft tissue sarcoma microenvironment.Human Pathol.2015;46:357-65.
28. Hou JY,Yu Z,Xiang RY,Li CQ,Wang L,Chen SF,et al.Correlation between infiltration of FOXP3+regulatory T cells and expression of B7-H1 in the tumor tissues of gastric cancer.Exp Mol Pathol.2014;96:284-91.
29. Deng WW,Mao L,Yu GT,Bu LL,Ma SR,Liu B,et al.LAG-3confers poor prognosis and its blockade reshapes antitumor response in head and neck squamous cell carcinoma.Oncoimmunology.2016;5:e1239005.
30. Chen DS,Mellman I.Oncology meets immunology:the cancerimmunity cycle.Immunity.2013;39:1-10.
31. Topalian SL,Drake CG,Pardoll DM.Immune checkpoint blockade:a common denominator approach to cancer therapy.Cancer Cell.2015;27:450-61.
32. Liu JF,Wu L,Yang LL,Deng WW,Mao L,Wu H,et al.Blockade ofTIM3 relieves immunosuppression through reducing regulatory T cells in head and neck cancer.J Exp Clin Cancer Res.2018;37:44.
33. Fridman WH,Pages F,Sautes-Fridman C,Galon J.The immune contexture in human tumours:impact on clinical outcome.Nat Rev Cancer.2012;12:298-306.
34. Muris JJ,Meijer CJ,Cillessen SA,Vos W,Kummer JA,Bladergroen BA,et al.Prognostic significance of activated cytotoxic Tlymphocytes in primary nodal diffuse large B-cell lymphomas.Leukemia.2004;18:589-96.
35. Scott DW,Chan FC,Hong FX,Rogic S,Tan KL,Meissner B,et al.Gene expression-based model using formalin-fixed paraffinembedded biopsies predicts overall survival in advanced-stage classical Hodgkin lymphoma.J Clin Oncol.2013;31:692-700.
36. Nakano O,Sato M,Naito Y,Suzuki K,Orikasa S,Aizawa M,et al.Proliferative activity of intratumoral CD8+T-lymphocytes as a prognostic factor in human renal cell carcinoma:clinicopathologic demonstration of antitumor immunity.Cancer Res.2001;61:5132-36.
37. Sorbye SW,Kilvaer T,Valkov A,Donnem T,Smeland E,Al-Shibli K,et al.Prognostic impact of lymphocytes in soft tissue sarcomas.PLoS One.2011;6:e14611.
38. Grosso JF,Kelleher CC,Harris TJ,Maris CH,Hipkiss EL,De Marzo A,et al.LAG-3 regulates CD8+T cell accumulation and effector function in murine self-and tumor-tolerance systems.J Clin Invest.2007;117:3383-92.
39. Hannier S,Triebel F.The MHC class II ligand lymphocyte activation gene-3 is co-distributed with CD8 and CD3-TCR molecules after their engagement by mAb or peptide-MHC class I complexes.Int Immunol.1999;11:1745-52.
40. Toulmonde M,Penel N,Adam J,Chevreau C,Blay JY,Le Cesne A.et al.Use of PD-1 targeting,macrophage infiltration,and IDO pathway activation in sarcomas:a phase 2 clinical trial.JAMA Oncol.2018;4:93-7.
41. D'Angelo SP,Shoushtari AN,Keohan ML,Dickson MA,Gounder MM,Chi P,et al.Combined KIT and CTLA-4 Blockade in Patients with Refractory GIST and other advanced sarcomas:a phase ib study of dasatinib plus Ipilimumab.Clin Cancer Res.2017;23:2972-80.
42. Triebel F,Jitsukawa S,Baixeras E,Roman-Roman S,Genevee C,Viegas-Pequignot E,et al.LAG-3,a novel lymphocyte activation gene closely related to CD4.J Exp Med.1990;171:1393-405.