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Selective inhibition of the p38 alternative activation pathway in infiltrating T cells inhibits pancreatic cancer progression

Abstract

Pancreatic ductal adenocarcinoma (PDAC) is a highly aggressive neoplasm characterized by a marked fibro-inflammatory microenvironment1, the presence of which can promote both cancer induction and growth2,3,4. Therefore, selective manipulation of local cytokines is an attractive, although unrealized, therapeutic approach. T cells possess a unique mechanism of p38 mitogen-activated protein kinase (MAPK) activation downstream of T cell receptor (TCR) engagement through the phosphorylation of Tyr323 (pY323). This alternative p38 activation pathway is required for pro-inflammatory cytokine production5,6. Here we show in human PDAC that a high percentage of infiltrating pY323+ T cells was associated with large numbers of tumor necrosis factor (TNF)-α− and interleukin (IL)-17–producing CD4+ tumor-infiltrating lymphocytes (TILs) and aggressive disease. The growth of mouse pancreatic tumors was inhibited by genetic ablation of the alternative p38 pathway, and transfer of wild-type CD4+ T cells, but not those lacking the alternative pathway, enhanced tumor growth in T cell–deficient mice. Notably, a plasma membrane–permeable peptide derived from GADD45-α, the naturally occurring inhibitor of p38 pY323+ (ref. 7), reduced CD4+ TIL production of TNF-α, IL-17A, IL-10 and secondary cytokines, halted growth of implanted tumors and inhibited progression of spontaneous KRAS-driven adenocarcinoma in mice. Thus, TCR-mediated activation of CD4+ TILs results in alternative p38 activation and production of protumorigenic factors and can be targeted for therapeutic benefit.

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Figure 1: A high percentage of p38 pY323+ TILs is associated with enhanced pro-inflammatory cytokine production and shorter survival in human PDAC.
Figure 2: CD4+ T cells activated by the p38 alternative pathway promote the growth of pancreatic cancer cells.
Figure 3: GADD45-α residues 71–85 bind p38 and inhibit T cell proliferation and effector function.
Figure 4: (11R)–GADD45-α71–85 treatment inhibits tumor growth, angiogenesis, PanIN progression and invasive cancer in mice.

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Acknowledgements

We thank J. Scheuerer, M. Clauter, B. Walter and E. Castro for expert technical assistance, J. Greiner for Panc02 cells, the Center for Advanced Preclinical Research (CAPR) program at the Frederick National Laboratory for Cancer Research for providing KPC mice for some studies, T. Longerich and B. Lahrmann for technical advice in computer-based tissue analysis and P. Schirmacher for critical review of the manuscript. This work was supported by the Intramural Research Program of the Center for Cancer Research, National Cancer Institute, National Institutes of Health. M.M.G. was supported by a fellowship from the German Research Foundation (DFG; Ga-1818/1-1). The work of F.L. was in part funded by the German Research Foundation (grant no. SFB 938/TP Z2).

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M.S.A. and M.M.G. contributed equally to this manuscript and are listed in alphabetical order. M.S.A., M.M.G. and J.D.A. designed the experiments, analyzed the data and wrote the manuscript. M.S.A. and J.D.A. developed the cell-permeable GADD45-α fragment. M.S.A. and M.M.G. performed all mouse and in vitro experiments. M.M.G. did the tissue evaluation and the computer-based tissue analysis. F.B. provided the human paraffin tissues, created the human tissue microarray, performed the corresponding immunohistochemistry, and evaluated tissue samples together with M.M.G. F.L. performed and analyzed multicolor-immunofluorescence staining of human tissues. T.G., N.A.G. and T.H. collected fresh-frozen tissues, performed qPCR and provided patient clinical data. S.P.H. and S.V.K. screened and provided KPC mice. S.V.K. and U.H. performed biostatistics and multivariate analysis of human data.

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Correspondence to Jonathan D Ashwell.

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Alam, M., Gaida, M., Bergmann, F. et al. Selective inhibition of the p38 alternative activation pathway in infiltrating T cells inhibits pancreatic cancer progression. Nat Med 21, 1337–1343 (2015). https://doi.org/10.1038/nm.3957

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