Limits...
Checkpoint blockade cancer immunotherapy targets tumour-specific mutant antigens.

Gubin MM, Zhang X, Schuster H, Caron E, Ward JP, Noguchi T, Ivanova Y, Hundal J, Arthur CD, Krebber WJ, Mulder GE, Toebes M, Vesely MD, Lam SS, Korman AJ, Allison JP, Freeman GJ, Sharpe AH, Pearce EL, Schumacher TN, Aebersold R, Rammensee HG, Melief CJ, Mardis ER, Gillanders WE, Artyomov MN, Schreiber RD - Nature (2014)

Bottom Line: Yet, clinically apparent cancers still arise in immunocompetent individuals in part as a consequence of cancer-induced immunosuppression.Monoclonal-antibody-based therapies targeting CTLA-4 and/or PD-1 (checkpoint blockade) have yielded significant clinical benefits-including durable responses--to patients with different malignancies.These results reveal that tumour-specific mutant antigens are not only important targets of checkpoint blockade therapy, but they can also be used to develop personalized cancer-specific vaccines and to probe the mechanistic underpinnings of different checkpoint blockade treatments.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Immunology, Washington University School of Medicine, 660 South Euclid Avenue, St Louis, Missouri 63110, USA.

ABSTRACT
The immune system influences the fate of developing cancers by not only functioning as a tumour promoter that facilitates cellular transformation, promotes tumour growth and sculpts tumour cell immunogenicity, but also as an extrinsic tumour suppressor that either destroys developing tumours or restrains their expansion. Yet, clinically apparent cancers still arise in immunocompetent individuals in part as a consequence of cancer-induced immunosuppression. In many individuals, immunosuppression is mediated by cytotoxic T-lymphocyte associated antigen-4 (CTLA-4) and programmed death-1 (PD-1), two immunomodulatory receptors expressed on T cells. Monoclonal-antibody-based therapies targeting CTLA-4 and/or PD-1 (checkpoint blockade) have yielded significant clinical benefits-including durable responses--to patients with different malignancies. However, little is known about the identity of the tumour antigens that function as the targets of T cells activated by checkpoint blockade immunotherapy and whether these antigens can be used to generate vaccines that are highly tumour-specific. Here we use genomics and bioinformatics approaches to identify tumour-specific mutant proteins as a major class of T-cell rejection antigens following anti-PD-1 and/or anti-CTLA-4 therapy of mice bearing progressively growing sarcomas, and we show that therapeutic synthetic long-peptide vaccines incorporating these mutant epitopes induce tumour rejection comparably to checkpoint blockade immunotherapy. Although mutant tumour-antigen-specific T cells are present in progressively growing tumours, they are reactivated following treatment with anti-PD-1 and/or anti-CTLA-4 and display some overlapping but mostly treatment-specific transcriptional profiles, rendering them capable of mediating tumour rejection. These results reveal that tumour-specific mutant antigens are not only important targets of checkpoint blockade therapy, but they can also be used to develop personalized cancer-specific vaccines and to probe the mechanistic underpinnings of different checkpoint blockade treatments.

Show MeSH

Related in: MedlinePlus

Detection of TIM-3, LAG-3, IFN-γ and TNF-α expression in tumour infiltrating CD8+ T cellsa, Representative histogram of TIM-3 or LAG-3 expression on mLama4-specific CD8+ tumour infiltrating T cells from tumour bearing mice treated with αPD-1, αCTLA-4, both αPD-1 and αCTLA-4 or control mAbs. b, TIM-3 and LAG-3 are reduced in mAlg8-specific CD8+ TILs from tumour-bearing mice treated with αPD-1, αCTLA-4, or both αPD-1 and αCTLA-4 compared to mice treated with control mAb. N=5 mice per group pooled. Data are presented as mean ± s.e.m. of at least three independent experiments. Samples were compared using an unpaired, two-tailed Student’s t test (*p<0.05, **p<0.01). c, Representative dot plots of IFN-γ and TNF-α stained CD8+ tumour-infiltrating T cells from tumour-bearing mice following treatment with αPD-1, αCTLA-4, both αPD-1 and αCTLA-4 or control mAbs. Data presented are representative of at least three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
getmorefigures.php?uid=PMC4279952&req=5

Figure 13: Detection of TIM-3, LAG-3, IFN-γ and TNF-α expression in tumour infiltrating CD8+ T cellsa, Representative histogram of TIM-3 or LAG-3 expression on mLama4-specific CD8+ tumour infiltrating T cells from tumour bearing mice treated with αPD-1, αCTLA-4, both αPD-1 and αCTLA-4 or control mAbs. b, TIM-3 and LAG-3 are reduced in mAlg8-specific CD8+ TILs from tumour-bearing mice treated with αPD-1, αCTLA-4, or both αPD-1 and αCTLA-4 compared to mice treated with control mAb. N=5 mice per group pooled. Data are presented as mean ± s.e.m. of at least three independent experiments. Samples were compared using an unpaired, two-tailed Student’s t test (*p<0.05, **p<0.01). c, Representative dot plots of IFN-γ and TNF-α stained CD8+ tumour-infiltrating T cells from tumour-bearing mice following treatment with αPD-1, αCTLA-4, both αPD-1 and αCTLA-4 or control mAbs. Data presented are representative of at least three independent experiments.

Mentions: We also assessed changes in expression of functionally relevant proteins on/in CD8+ TILs in mice undergoing treatment with different checkpoint blocking mAbs. TILs specific for mLama4 or mAlg8 from mice treated with αPD-1 and/or αCTLA-4 displayed lower cell surface expression of Lymphocyte-Activation Gene 3 (LAG-3) and T Cell Immunoglobulin and Mucin Protein 3 (TIM-3) than those in progressively growing tumours in control mAb treated mice (Fig. 4a and Extended Data Fig. 9a,b). Elevated LAG-3 and TIM-3 expression is known to mark antigen-experienced, dysfunctional (i.e., exhausted) CD8+ T cells16,17 in chronic viral infection. Conversely, TILs specific for mLama4 or mAlg8 from αCTLA-4 or αCTLA-4 + αPD-1 treated mice displayed significantly higher levels of Granzyme B than antigen-specific TILs from mice treated with either αPD-1 alone or control mAb (Fig. 4b). Consistent with the RNA-Seq analysis, these changes were observed predominantly in antigen-specific TILs. In addition, whereas a low percentage of CD8+ TILs from mice treated with control mAb produced IFN-γ and TNF-α (Fig. 4c and Extended Data Fig. 9c), the percentage of IFN-γ producing TILs increased following treatment of the mice with αPD-1 and particularly with αCTLA-4 or the combination of αCTLA-4 + αPD-1. TILs expressing both cytokines, which likely represent the most potent anti-tumour effectors, were most highly represented following treatment of tumour bearing mice with the combination of αCTLA-4 + αPD-1.


Checkpoint blockade cancer immunotherapy targets tumour-specific mutant antigens.

Gubin MM, Zhang X, Schuster H, Caron E, Ward JP, Noguchi T, Ivanova Y, Hundal J, Arthur CD, Krebber WJ, Mulder GE, Toebes M, Vesely MD, Lam SS, Korman AJ, Allison JP, Freeman GJ, Sharpe AH, Pearce EL, Schumacher TN, Aebersold R, Rammensee HG, Melief CJ, Mardis ER, Gillanders WE, Artyomov MN, Schreiber RD - Nature (2014)

Detection of TIM-3, LAG-3, IFN-γ and TNF-α expression in tumour infiltrating CD8+ T cellsa, Representative histogram of TIM-3 or LAG-3 expression on mLama4-specific CD8+ tumour infiltrating T cells from tumour bearing mice treated with αPD-1, αCTLA-4, both αPD-1 and αCTLA-4 or control mAbs. b, TIM-3 and LAG-3 are reduced in mAlg8-specific CD8+ TILs from tumour-bearing mice treated with αPD-1, αCTLA-4, or both αPD-1 and αCTLA-4 compared to mice treated with control mAb. N=5 mice per group pooled. Data are presented as mean ± s.e.m. of at least three independent experiments. Samples were compared using an unpaired, two-tailed Student’s t test (*p<0.05, **p<0.01). c, Representative dot plots of IFN-γ and TNF-α stained CD8+ tumour-infiltrating T cells from tumour-bearing mice following treatment with αPD-1, αCTLA-4, both αPD-1 and αCTLA-4 or control mAbs. Data presented are representative of at least three independent experiments.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4279952&req=5

Figure 13: Detection of TIM-3, LAG-3, IFN-γ and TNF-α expression in tumour infiltrating CD8+ T cellsa, Representative histogram of TIM-3 or LAG-3 expression on mLama4-specific CD8+ tumour infiltrating T cells from tumour bearing mice treated with αPD-1, αCTLA-4, both αPD-1 and αCTLA-4 or control mAbs. b, TIM-3 and LAG-3 are reduced in mAlg8-specific CD8+ TILs from tumour-bearing mice treated with αPD-1, αCTLA-4, or both αPD-1 and αCTLA-4 compared to mice treated with control mAb. N=5 mice per group pooled. Data are presented as mean ± s.e.m. of at least three independent experiments. Samples were compared using an unpaired, two-tailed Student’s t test (*p<0.05, **p<0.01). c, Representative dot plots of IFN-γ and TNF-α stained CD8+ tumour-infiltrating T cells from tumour-bearing mice following treatment with αPD-1, αCTLA-4, both αPD-1 and αCTLA-4 or control mAbs. Data presented are representative of at least three independent experiments.
Mentions: We also assessed changes in expression of functionally relevant proteins on/in CD8+ TILs in mice undergoing treatment with different checkpoint blocking mAbs. TILs specific for mLama4 or mAlg8 from mice treated with αPD-1 and/or αCTLA-4 displayed lower cell surface expression of Lymphocyte-Activation Gene 3 (LAG-3) and T Cell Immunoglobulin and Mucin Protein 3 (TIM-3) than those in progressively growing tumours in control mAb treated mice (Fig. 4a and Extended Data Fig. 9a,b). Elevated LAG-3 and TIM-3 expression is known to mark antigen-experienced, dysfunctional (i.e., exhausted) CD8+ T cells16,17 in chronic viral infection. Conversely, TILs specific for mLama4 or mAlg8 from αCTLA-4 or αCTLA-4 + αPD-1 treated mice displayed significantly higher levels of Granzyme B than antigen-specific TILs from mice treated with either αPD-1 alone or control mAb (Fig. 4b). Consistent with the RNA-Seq analysis, these changes were observed predominantly in antigen-specific TILs. In addition, whereas a low percentage of CD8+ TILs from mice treated with control mAb produced IFN-γ and TNF-α (Fig. 4c and Extended Data Fig. 9c), the percentage of IFN-γ producing TILs increased following treatment of the mice with αPD-1 and particularly with αCTLA-4 or the combination of αCTLA-4 + αPD-1. TILs expressing both cytokines, which likely represent the most potent anti-tumour effectors, were most highly represented following treatment of tumour bearing mice with the combination of αCTLA-4 + αPD-1.

Bottom Line: Yet, clinically apparent cancers still arise in immunocompetent individuals in part as a consequence of cancer-induced immunosuppression.Monoclonal-antibody-based therapies targeting CTLA-4 and/or PD-1 (checkpoint blockade) have yielded significant clinical benefits-including durable responses--to patients with different malignancies.These results reveal that tumour-specific mutant antigens are not only important targets of checkpoint blockade therapy, but they can also be used to develop personalized cancer-specific vaccines and to probe the mechanistic underpinnings of different checkpoint blockade treatments.

View Article: PubMed Central - PubMed

Affiliation: Department of Pathology and Immunology, Washington University School of Medicine, 660 South Euclid Avenue, St Louis, Missouri 63110, USA.

ABSTRACT
The immune system influences the fate of developing cancers by not only functioning as a tumour promoter that facilitates cellular transformation, promotes tumour growth and sculpts tumour cell immunogenicity, but also as an extrinsic tumour suppressor that either destroys developing tumours or restrains their expansion. Yet, clinically apparent cancers still arise in immunocompetent individuals in part as a consequence of cancer-induced immunosuppression. In many individuals, immunosuppression is mediated by cytotoxic T-lymphocyte associated antigen-4 (CTLA-4) and programmed death-1 (PD-1), two immunomodulatory receptors expressed on T cells. Monoclonal-antibody-based therapies targeting CTLA-4 and/or PD-1 (checkpoint blockade) have yielded significant clinical benefits-including durable responses--to patients with different malignancies. However, little is known about the identity of the tumour antigens that function as the targets of T cells activated by checkpoint blockade immunotherapy and whether these antigens can be used to generate vaccines that are highly tumour-specific. Here we use genomics and bioinformatics approaches to identify tumour-specific mutant proteins as a major class of T-cell rejection antigens following anti-PD-1 and/or anti-CTLA-4 therapy of mice bearing progressively growing sarcomas, and we show that therapeutic synthetic long-peptide vaccines incorporating these mutant epitopes induce tumour rejection comparably to checkpoint blockade immunotherapy. Although mutant tumour-antigen-specific T cells are present in progressively growing tumours, they are reactivated following treatment with anti-PD-1 and/or anti-CTLA-4 and display some overlapping but mostly treatment-specific transcriptional profiles, rendering them capable of mediating tumour rejection. These results reveal that tumour-specific mutant antigens are not only important targets of checkpoint blockade therapy, but they can also be used to develop personalized cancer-specific vaccines and to probe the mechanistic underpinnings of different checkpoint blockade treatments.

Show MeSH
Related in: MedlinePlus