Melanomas resist T-cell therapy through inflammation-induced reversible dedifferentiation

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Abstract

Adoptive cell transfer therapies (ACTs) with cytotoxic T cells that target melanocytic antigens can achieve remissions in patients with metastatic melanomas, but tumours frequently relapse1,2. Hypotheses explaining the acquired resistance to ACTs include the selection of antigen-deficient tumour cell variants3,4,5 and the induction of T-cell tolerance6. However, the lack of appropriate experimental melanoma models has so far impeded clear insights into the underlying mechanisms. Here we establish an effective ACT protocol in a genetically engineered mouse melanoma model that recapitulates tumour regression, remission and relapse as seen in patients. We report the unexpected observation that melanomas acquire ACT resistance through an inflammation-induced reversible loss of melanocytic antigens. In serial transplantation experiments, melanoma cells switch between a differentiated and a dedifferentiated phenotype in response to T-cell-driven inflammatory stimuli. We identified the proinflammatory cytokine tumour necrosis factor (TNF)-α as a crucial factor that directly caused reversible dedifferentiation of mouse and human melanoma cells. Tumour cells exposed to TNF-α were poorly recognized by T cells specific for melanocytic antigens, whereas recognition by T cells specific for non-melanocytic antigens was unaffected or even increased. Our results demonstrate that the phenotypic plasticity of melanoma cells in an inflammatory microenvironment contributes to tumour relapse after initially successful T-cell immunotherapy. On the basis of our work, we propose that future ACT protocols should simultaneously target melanocytic and non-melanocytic antigens to ensure broad recognition of both differentiated and dedifferentiated melanoma cells, and include strategies to sustain T-cell effector functions by blocking immune-inhibitory mechanisms in the tumour microenvironment.

Figure 1: HGF–CDK4(R24C) melanomas resisting ACT with gp100-specific CTLs show partial loss of antigen expression in an inflammatory tumour microenvironment.
Figure 2: Loss of gp100 antigen expression in relapsed HGF–CDK4(R24C) melanoma lines is reversible after re-transplantation.
Figure 3: Proinflammatory mediators directly promote HGF–CDK4(R24C) melanoma cell dedifferentiation leading to impaired recognition by gp100-specific CTLs.
Figure 4: TNF-α induces human melanoma cell dedifferentiation leading to selectively impaired recognition by autologous CTLs specific for melanocytic antigens.

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Primary accessions

Gene Expression Omnibus

Data deposits

The microarray data have been deposited in the Gene Expression Omnibus (GEO) database under the accession number GSE40213.

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Acknowledgements

We would like to thank G. Merlino and M. Barbacid for providing genetically engineered mice; E. Endl for help with FACS sorting; S. Herms and P. Hoffmann for the cDNA microarrays; S. Mikus, A. Sporleder and C. Lemke for managing the mouse colony and performing histopathology, immunohistopatholgoy, immunoblotting and real-time PCR. This research was supported in part by grants from the DFG (A12 in the SFB832 and A22 in the SFB704) and the Deutsche Krebshilfe (P9 in the Melanoma Research Network) to T.T., by grants from BONFOR to J.L. and J.K., and by a DFG grant (A1 in the SFB 432) to T.W. T.T. and M.H. are members of the Excellence Cluster ImmunoSensation.

Author information

J.L., J.K., M.Re., T.B. and M.C. performed all experiments with mouse melanomas, analysed the data and reviewed the manuscript. J.K., M.Ro. and M.F. performed experiments with human melanoma cells and analysed the data. V.L. and T.W. designed and supervised experiments with human melanoma cells, interpreted the data and reviewed the manuscript. M.H. designed experiments, performed the bioinformatic analyses, interpreted data and helped to write the manuscript. T.T. conceived and supervised all aspects of the project, designed experiments, interpreted the data and wrote the manuscript.

Correspondence to Thomas Tüting.

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Landsberg, J., Kohlmeyer, J., Renn, M. et al. Melanomas resist T-cell therapy through inflammation-induced reversible dedifferentiation. Nature 490, 412–416 (2012) doi:10.1038/nature11538

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