Letter | Published:

Melanoma-intrinsic β-catenin signalling prevents anti-tumour immunity

Nature volume 523, pages 231235 (09 July 2015) | Download Citation

Abstract

Melanoma treatment is being revolutionized by the development of effective immunotherapeutic approaches1,2. These strategies include blockade of immune-inhibitory receptors on activated T cells; for example, using monoclonal antibodies against CTLA-4, PD-1, and PD-L1 (refs 3, 4, 5). However, only a subset of patients responds to these treatments, and data suggest that therapeutic benefit is preferentially achieved in patients with a pre-existing T-cell response against their tumour, as evidenced by a baseline CD8+ T-cell infiltration within the tumour microenvironment6,7. Understanding the molecular mechanisms that underlie the presence or absence of a spontaneous anti-tumour T-cell response in subsets of cases, therefore, should enable the development of therapeutic solutions for patients lacking a T-cell infiltrate. Here we identify a melanoma-cell-intrinsic oncogenic pathway that contributes to a lack of T-cell infiltration in melanoma. Molecular analysis of human metastatic melanoma samples revealed a correlation between activation of the WNT/β-catenin signalling pathway and absence of a T-cell gene expression signature. Using autochthonous mouse melanoma models8,9 we identified the mechanism by which tumour-intrinsic active β-catenin signalling results in T-cell exclusion and resistance to anti-PD-L1/anti-CTLA-4 monoclonal antibody therapy. Specific oncogenic signals, therefore, can mediate cancer immune evasion and resistance to immunotherapies, pointing to new candidate targets for immune potentiation.

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Primary accessions

Gene Expression Omnibus

Data deposits

Gene array data have been deposited in the Gene Expression Omnibus under accession number GSE63543.

References

  1. 1.

    et al. The Society for Immunotherapy of Cancer consensus statement on tumour immunotherapy for the treatment of cutaneous melanoma. Nature Rev. Clin. Oncol. 10, 588–598 (2013)

  2. 2.

    , & Cancer immunotherapy comes of age. Nature 480, 480–489 (2011)

  3. 3.

    et al. Nivolumab plus ipilimumab in advanced melanoma. N. Engl. J. Med. 369, 122–133 (2013)

  4. 4.

    et al. Survival, durable tumor remission, and long-term safety in patients with advanced melanoma receiving nivolumab. J. Clin. Oncol. 32, 1020–1030 (2014)

  5. 5.

    et al. Improved survival with ipilimumab in patients with metastatic melanoma. N. Engl. J. Med. 363, 711–723 (2010)

  6. 6.

    et al. Chemokine expression in melanoma metastases associated with CD8+ T-cell recruitment. Cancer Res. 69, 3077–3085 (2009)

  7. 7.

    et al. An immune-active tumor microenvironment favors clinical response to ipilimumab. Cancer Immunol. Immunother. 61, 1019–1031 (2012)

  8. 8.

    et al. Braf(V600E) cooperates with Pten loss to induce metastatic melanoma. Nature Genet. 41, 544–552 (2009)

  9. 9.

    et al. β-Catenin signaling controls metastasis in Braf-activated Pten-deficient melanomas. Cancer Cell 20, 741–754 (2011)

  10. 10.

    et al. Type, density, and location of immune cells within human colorectal tumors predict clinical outcome. Science 313, 1960–1964 (2006)

  11. 11.

    , , , & Frequent nuclear/cytoplasmic localization of β-catenin without exon 3 mutations in malignant melanoma. Am. J. Pathol. 154, 325–329 (1999)

  12. 12.

    et al. Up-regulation of PD-L1, IDO, and Tregs in the melanoma tumor microenvironment is driven by CD8+ T cells. Sci. Transl. Med. 5, 200ra116 (2013)

  13. 13.

    et al. Immune inhibitory molecules LAG-3 and PD-1 synergistically regulate T-cell function to promote tumoral immune escape. Cancer Res. 72, 917–927 (2012)

  14. 14.

    et al. Melanomas resist T-cell therapy through inflammation-induced reversible dedifferentiation. Nature 490, 412–416 (2012)

  15. 15.

    et al. Cancer exome analysis reveals a T-cell-dependent mechanism of cancer immunoediting. Nature 482, 400–404 (2012)

  16. 16.

    , , , & Regulated expression of a tumor-associated antigen reveals multiple levels of T-cell tolerance in a mouse model of lung cancer. Cancer Res. 68, 9459–9468 (2008)

  17. 17.

    et al. Host type I IFN signals are required for antitumor CD8+ T cell responses through CD8α+ dendritic cells. J. Exp. Med. 208, 2005–2016 (2011)

  18. 18.

    et al. Batf3 deficiency reveals a critical role for CD8α+ dendritic cells in cytotoxic T cell immunity. Science 322, 1097–1100 (2008)

  19. 19.

    et al. Cross-presentation of viral and self antigens by skin-derived CD103+ dendritic cells. Nature Immunol. 10, 488–495 (2009)

  20. 20.

    et al. Flt3L dependence helps define an uncharacterized subset of murine cutaneous dendritic cells. J. Invest. Dermatol. 134, 1265–1275 (2014)

  21. 21.

    et al. CCR5 provides a signal for microbial induced production of IL-12 by CD8α+ dendritic cells. Nature Immunol. 1, 83–87 (2000)

  22. 22.

    et al. PD-1 blockade enhances T-cell migration to tumors by elevating IFN-γ inducible chemokines. Cancer Res. 72, 5209–5218 (2012)

  23. 23.

    et al. N-myc downstream-regulated gene 2, a novel estrogen-targeted gene, is involved in the regulation of Na+/K+-ATPase. J. Biol. Chem. 286, 32289–32299 (2011)

  24. 24.

    , , & Activating transcription factor 3 (ATF3) represses the expression of CCL4 in murine macrophages. Mol. Immunol. 44, 1598–1605 (2007)

  25. 25.

    et al. Human CD141+ (BDCA-3)+ dendritic cells (DCs) represent a unique myeloid DC subset that cross-presents necrotic cell antigens. J. Exp. Med. 207, 1247–1260 (2010)

  26. 26.

    et al. Mechanism of tumor rejection with doublets of CTLA-4, PD-1/PD-L1, or IDO blockade involves restored IL-2 production and proliferation of CD8+ T cells directly within the tumor microenvironment. J. Immunother. Cancer (2014)

  27. 27.

    et al. Immune suppression and resistance mediated by constitutive activation of Wnt/β-catenin signaling in human melanoma cells. J. Immunol. 189, 2110–2117 (2012)

  28. 28.

    et al. β-Catenin inhibits T cell activation by selective interference with linker for activation of T cells–phospholipase C-γ1 phosphorylation. J. Immunol. 186, 784–790 (2011)

  29. 29.

    , , & Tumor-infiltrating lymphocytes: apparently good for melanoma patients. But why? Cancer Immunol. Immunother. 60, 1153–1160 (2011)

  30. 30.

    & RSEM: accurate transcript quantification from RNA-Seq data with or without a reference genome. BMC Bioinformatics 12, 323 (2011)

  31. 31.

    & ConsensusClusterPlus: a class discovery tool with confidence assessments and item tracking. Bioinformatics 26, 1572–1573 (2010)

  32. 32.

    , & ANNOVAR: functional annotation of genetic variants from high-throughput sequencing data. Nucleic Acids Res. 38, e164 (2010)

  33. 33.

    The 1000 Genomes Project Consortium. An integrated map of genetic variation from 1,092 human genomes. Nature 491, 56–65 (2012)

  34. 34.

    et al. STRING 8—a global view on proteins and their functional interactions in 630 organisms. Nucleic Acids Res. 37, D412–D416 (2009)

  35. 35.

    et al. High cancer susceptibility and embryonic lethality associated with mutation of the PTEN tumor suppressor gene in mice. Curr. Biol. 8, 1169–1178 (1998)

  36. 36.

    et al. Stabilization of β-catenin induces lesions reminiscent of prostatic intraepithelial neoplasia, but terminal squamous transdifferentiation of other secretory epithelia. Oncogene 21, 4099–4107 (2002)

  37. 37.

    et al. A new mouse model to explore the initiation, progression, and therapy of BRAFV600E-induced lung tumors. Genes Dev. 21, 379–384 (2007)

  38. 38.

    et al. Characterization of melanocyte-specific inducible Cre recombinase transgenic mice. Genesis 44, 262–267 (2006)

  39. 39.

    , , , & Hedgehog signaling in the neural crest cells regulates the patterning and growth of facial primordia. Genes Dev. 18, 937–951 (2004)

  40. 40.

    et al. Antigen recognition and allogeneic tumor rejection in CD8+ TCR transgenic/RAG−/− mice. J. Immunol. 159, 4665–4675 (1997)

  41. 41.

    et al. Immunotype and immunohistologic characteristics of tumor-infiltrating immune cells are associated with clinical outcome in metastatic melanoma. Cancer Res. 72, 1070–1080 (2012)

  42. 42.

    & Analyzing real-time PCR data by the comparative CT method. Nature Protocols 3, 1101–1108 (2008)

  43. 43.

    et al. Generation of Th1-polarizing dendritic cells using the TLR7/8 agonist CL075. J. Immunol. 185, 738–747 (2010)

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Acknowledgements

The authors would like to thank A. Sailer and J. Turner for their assistance on mouse tissue immunofluorescent staining, M. Leung and Y. Zha for technical support, and the Special Services Animal Resources Center for assistance with mouse husbandry. We also acknowledge the Fitch Monoclonal Antibody Facility, the Human Tissue Research Core and the Integrated Microscopy core of The University of Chicago Comprehensive Cancer Center. We would like to thank A. O. Emmanuel and F. Gounari for assistance with the ChIP assay as well as for conditional β-catenin knock-in mice; C. Slingluff, D. Deacon, J. Schaefer, G. Erdag and the University of Virginia Biorepository and Tissue Research Facility for melanoma biopsy specimens, and P. Savage for critical comments. Funding for this study was provided by a Team Science Award from the Melanoma Research Alliance and a Translational Research Grant from the Cancer Research Institute. S.S. was supported by the German Research Foundation and is currently a fellow of the Cancer Research Institute.

Author information

Affiliations

  1. Department of Pathology, The University of Chicago, Chicago, Illinois 60637, USA

    • Stefani Spranger
    •  & Thomas F. Gajewski
  2. Center for Research Informatics, The University of Chicago, Chicago, Illinois 60637, USA

    • Riyue Bao
  3. Department of Medicine, The University of Chicago, Chicago, Illinois 60637, USA

    • Thomas F. Gajewski

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Contributions

S.S. contributed to the overall project design, planned and performed experiments, and performed data analysis. R.B. performed analysis of the TCGA data set. T.F.G. designed the overall project. S.S. and T.F.G. wrote the manuscript.

Corresponding author

Correspondence to Thomas F. Gajewski.

Extended data

Supplementary information

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  1. 1.

    Supplementary Information

    This file contains Supplementary Tables 1-5 comprising:1 (a) Expression of T cell genes in segregated groups (b) Gene clusters after supervised hierarchical clustering (c) List of differentially expressed genes; 2 - Pearson correlation of β-catenin target genes and CD8a transcripts; 3 - Mutation analysis summary with (a) T cell signature low patients and (b) T cell signature high patients (c) Table summarizing potential pathway activators in patients with an active β-catenin signature; 4 - Mouse gene array data of differentially expressed genes (a) and a summary table focusing on chemokine expression (b); 5 - Detailed primer and antibody information (a) Genotyping primers (b) antibodies (c) qPCR primer/ probes (d) ChIP assay primer and (e) siRNA oligos.

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DOI

https://doi.org/10.1038/nature14404

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