American Association for Cancer Research
00085472can210164-sup-259087_3_supp_7371156_qybmzw.xlsx (806.58 kB)

Supplementary Files 1-11 from Protein Signatures of NK Cell–Mediated Melanoma Killing Predict Response to Immunotherapies

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posted on 2023-03-31, 04:24 authored by Sabrina Cappello, Hsu-Min Sung, Christian Ickes, Christine S. Gibhardt, Adina Vultur, Hilal Bhat, Zhongwen Hu, Patricia Brafford, Andreas Denger, Ioana Stejerean-Todoran, Rixa-Mareike Köhn, Verena Lorenz, Nicolas Künzel, Gabriela Salinas, Hedwig Stanisz, Tobias Legler, Peter Rehling, Michael P. Schön, Karl S. Lang, Volkhard Helms, Meenhard Herlyn, Markus Hoth, Carsten Kummerow, Ivan Bogeski

A file containing RPPA and RNAseq datasets, pathway analyses and prediction calculations presented in the main figures.


German Research Foundation



Despite impressive advances in melanoma-directed immunotherapies, resistance is common and many patients still succumb to metastatic disease. In this context, harnessing natural killer (NK) cells, which have thus far been sidelined in the development of melanoma immunotherapy, could provide therapeutic benefits for cancer treatment. To identify molecular determinants of NK cell–mediated melanoma killing (NKmK), we quantified NK-cell cytotoxicity against a panel of genetically diverse melanoma cell lines and observed highly heterogeneous susceptibility. Melanoma protein microarrays revealed a correlation between NKmK and the abundance and activity of a subset of proteins, including several metabolic factors. Oxidative phoshorylation, measured by oxygen consumption rate, negatively correlated with melanoma cell sensitivity toward NKmK, and proteins involved in mitochondrial metabolism and epithelial–mesenchymal transition were confirmed to regulate NKmK. Two- and three-dimensional killing assays and melanoma xenografts established that the PI3K/AKT/mTOR signaling axis controls NKmK via regulation of NK cell–relevant surface proteins. A “protein-killing-signature” based on the protein analysis predicted NKmK of additional melanoma cell lines and the response of patients with melanoma to anti-PD-1 checkpoint therapy. Collectively, these findings identify novel NK cell–related prognostic biomarkers and may contribute to improved and personalized melanoma-directed immunotherapies. NK-cell cytotoxicity assays and protein microarrays reveal novel biomarkers of NK cell–mediated melanoma killing and enable development of signatures to predict melanoma patient responsiveness to immunotherapies.