Tyrosine kinase pathways modulate tumor susceptibility to natural killer cells
Roberto Bellucci, Hong-Nam Nguyen, Allison Martin, Stefan Heinrichs, Anna C. Schinzel, William C. Hahn, Jerome Ritz
Roberto Bellucci, Hong-Nam Nguyen, Allison Martin, Stefan Heinrichs, Anna C. Schinzel, William C. Hahn, Jerome Ritz
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Research Article Oncology

Tyrosine kinase pathways modulate tumor susceptibility to natural killer cells

Abstract

Natural killer (NK) cells are primary effectors of innate immunity directed against transformed tumor cells. In response, tumor cells have developed mechanisms to evade NK cell–mediated lysis through molecular mechanisms that are not well understood. In the present study, we used a lentiviral shRNA library targeting more than 1,000 human genes to identify 83 genes that promote target cell resistance to human NK cell–mediated killing. Many of the genes identified in this genetic screen belong to common signaling pathways; however, none of them have previously been known to modulate susceptibility of human tumor cells to immunologic destruction. Gene silencing of two members of the JAK family (JAK1 and JAK2) increased the susceptibility of a variety of tumor cell types to NK-mediated lysis and induced increased secretion of IFN-γ by NK cells. Treatment of tumor cells with JAK inhibitors also increased susceptibility to NK cell activity. These findings may have important clinical implications and suggest that small molecule inhibitors of tyrosine kinases being developed as therapeutic antitumor agents may also have significant immunologic effects in vivo.

Authors

Roberto Bellucci, Hong-Nam Nguyen, Allison Martin, Stefan Heinrichs, Anna C. Schinzel, William C. Hahn, Jerome Ritz

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Figure 10

Top 34 differentially expressed genes in IM-9-JAK1-KO compared with control IM-9 cells.

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Top 34 differentially expressed genes in IM-9-JAK1-KO compared with cont...
(A) Gene expression of 2 independent experiments (#1 and #2) using the Gene 1.0 ST Array. Samples from IM-9 cell lines expressing 2 JAK1 shRNAs (Jak1-1 and Jak1-3) were compared with IM-9 cells expressing an irrelevant shRNA (shCTRL-2) and with IM-9 parental cells. Top-scoring genes were defined by a minimal fold-change of 1.5 and maximal q value of 0.4. (B) IM-9-JAK1, JAK2-KO, and control cells were cultured for 12 hours and CXCL10 in culture supernatant was measured using an ELISA assay. Data represent the mean values ± SEM of 3 independent experiments run in triplicate. *P < 0.05 compared with IM-9 cells transduced with an irrelevant shRNA. (C) IM-9-JAK1-KO, IM-9-JAK2-KO, IM-9, and IM-9 cells transduced with an irrelevant shRNA (shCTRL-2) were stained with an anti–TRAIL-R1–PE antibody and analyzed by flow cytometry. (D) IM-9-JAK1-KO, IM-9-JAK2-KO, and IM-9-shCTRL-2 cells were co-incubated with NKL at a 1:1 E/T ratio with or without CXCL10 blocking antibody for 12 hours. Data represent the mean values + SEM of IFN-γ secreted by NKL cells in 4 independent experiments. (E) IM-9-JAK1-KO, IM-9-JAK2-KO, and IM-9-shCTRL-2 cells were co-incubated with NKL at a 1:1 E/T ratio with or without TRAIL-R1–Fc chimera for 12 hours. Data represent the mean values + SEM of IFN-γ secreted by NKL cells in 3 independent experiments. *P < 0.05, **P < 0.01.