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Neutrophil elastase–mediated degradation of IRS-1 accelerates lung tumor growth

Abstract

Lung cancer is the leading cause of cancer death worldwide1. Recent data suggest that tumor-associated inflammatory cells may modify lung tumor growth and invasiveness2,3. To determine the role of neutrophil elastase (encoded by Elane) on tumor progression, we used the loxP-Stop-loxP K-rasG12D (LSL–K-ras) model of mouse lung adenocarcinoma4 to generate LSL–K-ras-Elane−/− mice. Tumor burden was markedly reduced in LSL–K-ras-Elane−/− mice at all time points after induction of mutant K-ras expression. Kaplan-Meier survival analysis showed that whereas all LSL–K-ras-Elane+/+ mice died, none of the mice lacking neutrophil elastase died. Neutrophil elastase directly induced tumor cell proliferation in both human and mouse lung adenocarcinomas by gaining access to an endosomal compartment within tumor cells, where it degraded insulin receptor substrate-1 (IRS-1). Immunoprecipitation studies showed that, as neutrophil elastase degraded IRS-1, there was increased interaction between phosphatidylinositol 3-kinase (PI3K) and the potent mitogen platelet-derived growth factor receptor (PDGFR), thereby skewing the PI3K axis toward tumor cell proliferation. The inverse relationship identified between neutrophil elastase and IRS-1 in LSL–K-ras mice was also identified in human lung adenocarcinomas, thus translating these findings to human disease. This study identifies IRS-1 as a key regulator of PI3K within malignant cells. Additionally, to our knowledge, this is the first description of a secreted proteinase gaining access to the inside of a cell and altering intracellular signaling.

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Figure 1: Neutrophil elastase promotes lung tumor growth in vivo.
Figure 2: Neutrophil elastase induces tumor cell proliferation.
Figure 3: Neutrophil elastase–induced proliferation is dependent upon PDGFR-PI3K signaling.
Figure 4: Neutrophil elastase colocalizes with and degrades IRS-1.

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Acknowledgements

This work was supported by grants K08HL085286 and RO1HL054853 from the US National Institutes of Health National Heart, Lung, and Blood Institute (A.M.H. and S.D.S.) and by grant RO1DK058259 from the US National Institutes of Health National Institute of Diabetes and Digestive and Kidney Diseases (S.J.F.). ONO Pharmaceutical provided the ONO-5046. LSL–K-ras mice were provided by T. Jacks (Massachusetts Institute of Technology). AdenoCre was obtained from the University of Iowa Gene Transfer Vector Core. We would like to recognize members of the Shapiro lab for their assistance and comments regarding the preparation of this manuscript.

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A.M.H. and S.D.S. designed the study and interpreted all data. A.M.H., D.M.R., H.J., A.D.G., E.E.E., H.E.M., D.B.S., L.A.M., C.R.K., K.M.J., K.A.B. and M.M. performed the experiments. S.R.L. performed statistical analyses. S.J.F. provided the IRS-1 expression vector and assisted with data interpretation. K.K.W. participated in study design and data interpretation. A.M.H. wrote the manuscript.

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Correspondence to A McGarry Houghton.

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The authors declare no competing financial interests.

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Supplementary Figures 1–4, Supplementary Table 1 and Supplementary Methods (PDF 1104 kb)

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Houghton, A., Rzymkiewicz, D., Ji, H. et al. Neutrophil elastase–mediated degradation of IRS-1 accelerates lung tumor growth. Nat Med 16, 219–223 (2010). https://doi.org/10.1038/nm.2084

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