Silencing of Irf7 pathways in breast cancer cells promotes bone metastasis through immune escape

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Abstract

Breast cancer metastasis is a key determinant of long-term patient survival. By comparing the transcriptomes of primary and metastatic tumor cells in a mouse model of spontaneous bone metastasis, we found that a substantial number of genes suppressed in bone metastases are targets of the interferon regulatory factor Irf7. Restoration of Irf7 in tumor cells or administration of interferon led to reduced bone metastases and prolonged survival time. In mice deficient in the interferon (IFN) receptor or in natural killer (NK) and CD8+ T cell responses, metastasis was accelerated, indicating that Irf7-driven suppression of metastasis was reliant on IFN signaling to host immune cells. We confirmed the clinical relevance of these findings in over 800 patients in which high expression of Irf7-regulated genes in primary tumors was associated with prolonged bone metastasis–free survival. This gene signature may identify patients that could benefit from IFN-based therapies. Thus, we have identified an innate immune pathway intrinsic to breast cancer cells, the suppression of which restricts immunosurveillance to enable metastasis.

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Figure 1: Bone metastases derived from breast carcinomas downregulate interferon pathways and other immune-related genes.
Figure 2: Expression of the transcription factor Irf7 is suppressed in bone metastases.
Figure 3: Restoration of Irf7 expression enhances IFN signaling and inhibits metastasis.
Figure 4: Modulation of the immune system by metastatic tumor cells and reversion of these effects by enforced expression of the type I IFN pathway.
Figure 5: IRF7 expression in human breast cancer tissues.
Figure 6: The effect of type I IFN treatment on metastasis.

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Acknowledgements

This work was supported by the Cancer Council Victoria (B.S.P.), the Australian National Health and Medical Research Council (NHMRC) (P.J.H.), the Association of International Cancer Research (AICR) (A.M.), the Operational Infrastructure Scheme of the Victorian State government Department of Business and Innovation, the Australian Research Council (ARC) Centre for Structural and Functional Microbial Genomics and fellowship support for B.S.P. (NHMRC), R.L.A. (Australian National Breast Cancer Foundation, NBCF), A.M. (NBCF), C.Y.S. (NBCF, Cure Cancer Australia). We thank P. Hill (St Vincent's Pathology, St Vincent's Hospital, Fitzroy, Australia) for providing archived human breast cancer tissues assistance in the pathological assessment of immunohistochemistry staining and B. Haibe-Kains for assistance with codes in R. We also thank F. Miller (Karmanos Cancer Institute, Detroit, MI) for providing 66cl4 and 4T1 cell lines.

Author information

B.N.B. and C.Y.S. generated the majority of data in the manuscript. N.P.W. and A.M. contributed to mouse experiments, including intramammary fatpad injections and mouse tissue harvests. Y.C., D.A. and N.E.M. carried out the fluorescence-activated cell sorting (FACS) analysis on immune populations. T.M. did the methylation-sensitive high-resolution melting (MS-HRM), S.F. (PhD student and graduate of computer science and genetics graduate, Monash University) and S.A.S. (bioinformatician) carried out the bioinformatics analysis of the mouse microarray experiments and promoter analyses. S.L., a clinician scientist with expertise in bioinformatics and biostatistics, carried out the prognostic analyses. P.A. is a pathologist who provided and scored the matched primary tumor and metastases tissue arrays. N.A.d.W. produced, purified and tested IFN-α1 for in vivo experiments. J.G. performed some microarray experiments. P.J.H. and B.S.P. were responsible for project design, interpretation of data and drafting the manuscript, in collaboration with R.L.A. and M.J.S.

Correspondence to Paul J Hertzog or Belinda S Parker.

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Bidwell, B., Slaney, C., Withana, N. et al. Silencing of Irf7 pathways in breast cancer cells promotes bone metastasis through immune escape. Nat Med 18, 1224–1231 (2012) doi:10.1038/nm.2830

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