Melanoma NOS1 expression promotes dysfunctional IFN signaling
Qiuzhen Liu, … , Ena Wang, Francesco M. Marincola
Qiuzhen Liu, … , Ena Wang, Francesco M. Marincola
Published April 1, 2014
Citation Information: J Clin Invest. 2014;124(5):2147-2159. https://doi.org/10.1172/JCI69611.
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Research Article Oncology

Melanoma NOS1 expression promotes dysfunctional IFN signaling

Abstract

In multiple forms of cancer, constitutive activation of type I IFN signaling is a critical consequence of immune surveillance against cancer; however, PBMCs isolated from cancer patients exhibit depressed STAT1 phosphorylation in response to IFN-α, suggesting IFN signaling dysfunction. Here, we demonstrated in a coculture system that melanoma cells differentially impairs the IFN-α response in PBMCs and that the inhibitory potential of a particular melanoma cell correlates with NOS1 expression. Comparison of gene transcription and array comparative genomic hybridization (aCGH) between melanoma cells from different patients indicated that suppression of IFN-α signaling correlates with an amplification of the NOS1 locus within segment 12q22-24. Evaluation of NOS1 levels in melanomas and IFN responsiveness of purified PBMCs from patients indicated a negative correlation between NOS1 expression in melanomas and the responsiveness of PBMCs to IFN-α. Furthermore, in an explorative study, NOS1 expression in melanoma metastases was negatively associated with patient response to adoptive T cell therapy. This study provides a link between cancer cell phenotype and IFN signal dysfunction in circulating immune cells.

Authors

Qiuzhen Liu, Sara Tomei, Maria Libera Ascierto, Valeria De Giorgi, Davide Bedognetti, Cuilian Dai, Lorenzo Uccellini, Tara Spivey, Zoltan Pos, Jaime Thomas, Jennifer Reinboth, Daniela Murtas, Qianbing Zhang, Lotfi Chouchane, Geoffrey R. Weiss, Craig L. Slingluff Jr., Peter P. Lee, Steven A. Rosenberg, Harvey Alter, Kaitai Yao, Ena Wang, Francesco M. Marincola

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Figure 1

Modulation of IFN-α-p-STAT1 in PBMCs by melanoma cell lines.

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Modulation of IFN-α-p-STAT1 in PBMCs by melanoma cell lines. (A) Top lef...
(A) Top left: Histograms of p-STAT1 levels in 25 melanoma cell lines. Isotype, basal, and IFN-α-p-STAT1 are displayed in the top, middle, and bottom panels to exemplify IFN-α-p-STAT1 variability. Bottom left: Transwell coculture of melanoma cells and PBMCs. Right: IFN-α-p-STAT1 (top) and basal p-STAT1 (bottom) in CD4+, CD8+, and monocyte subsets of PBMCs from 4 donors in triplicate experiments after a 7-day coculture with 12 melanoma cell lines (blue bar) or alone (Mono; red bar). (B) Top: Average IFN-α-p-STAT1 levels in CD4+, CD8+, and monocyte subsets from 4 donors cocultured with 12 melanoma cell lines or alone, as shown in A. Cocultured results were ranked according to IFN-α-p-STAT1 levels (*P < 0.05, **P < 0.005, and ***P < 0.0005, Wilcoxon test): 5 cell lines with strong inhibitory effects (reduction of IFN-α-p-STAT1 by 50% compared with PBMCs cultured alone) were determined to be L-mels, and the rest H-mels. Bottom: IFN-α-p-STAT1 in PBMC subsets cocultured with L-mels or H-mels (P = 0.0005, Wilcoxon test). (C) Top: Average IFN-α-p-STAT1 levels in L-mels before and after coculture were lower than those in H-mel cell lines (Mann-Whitney U test, P = 0.048 and 0.018) before and after coculture with PBMCs (shown for individual cell lines at the bottom). IFN-α-p-STAT1 was enhanced significantly after coculture with PBMCs only in H-mels (P = 0.047, Wilcoxon test). (D) IFN-α-p-STAT1 in melanoma cells correlated with the IFN-α-p-STAT1 in respective cocultures of CD4+, CD8+, and monocyte subsets (Spearman’s correlation).