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A gene signature of bone metastatic colonization sensitizes for tumor-induced osteolysis and predicts survival in lung cancer

Oncogene volume 33, pages 5090–5099 (2014)Cite this article

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Abstract

Bone metastasis of lung adenocarcinoma (AC) is a frequent complication of advanced disease. The purpose of this study was to identify key mediators conferring robust prometastatic activity with clinical significance. We isolated highly metastatic subpopulations (HMS) using a previously described in vivo model of lung AC bone metastasis. We performed transcriptomic profiling of HMS and stringent bioinformatics filtering. Functional validation was assessed by overexpression and lentiviral silencing of single, double and triple combination in vivo and in vitro. We identified HDAC4, PITX1 and ROBO1 that decreased bone metastatic ability after their simultaneous abrogation. These effects were solely linked to defects in osseous colonization. The molecular mechanisms related to bone colonization were mediated by non-cell autonomous effects that include the following: (1) a marked decrease in osteoclastogenic activity in vitro and in vivo, an effect associated with reduced pro-osteoclastogenic cytokines IL-11 and PTHrP expression levels, as well as decreased in vitro expression of stromal rankl in conditions mimicking tumor–stromal interactions; (2) an abrogated response to TGF-β signaling by decreased phosphorylation and levels of Smad2/3 in tumor cells and (3) an impaired metalloproteolytic activity in vitro. Interestingly, coexpression of HDAC4 and PITX1 conferred high prometastatic activity in vivo. Further, levels of both genes correlated with patients at higher risk of metastasis in a clinical lung AC data set and with a poorer clinical outcome. These findings provide functional and clinical evidence that this metastatic subset is an important determinant of osseous colonization. These data suggest novel therapeutic targets to effectively block lung AC bone metastasis.

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Abbreviations

AC:

adenocarcinoma

BLI:

bioluminescence imaging

CM:

conditioned medium

i.c.:

intracardiac

i.t.:

intratibial

KD:

knockdown

MMP:

matrix metalloprotease

SCDC:

single-cell-derived colonies

TRAP:

Tartrate-resistant acid phosphatase

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Acknowledgements

We are grateful to the Core Histology Unit and Imaging Group facilities for their outstanding technical assistance. We also thank the Proteomics, Genomics and Bioinformatics Unit at the CIMA. This work was supported by ‘UTE project FIMA’ agreement, The Cancer Research Thematic Network of the Health Institute Carlos III (RTICC RD06/0020/0066), PI042282, SAF-2009–11280, SAF2012-40056, grants 67/2005 and 09/2009 from the Government of Navarra, and ‘La Caixa Foundation’ to FL. DL-R was supported by the FIMA and FPU. IA was funded by the Basque Government. JDLR was supported by FIS-ISCIII grant PI12/00624. SV is an investigator of the Ramon y Cajal Program (MICINN, RYC-2011-09042).

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Authors and Affiliations

  1. Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain

    D Luis-Ravelo, I Antón, C Zandueta, K Valencia, C Ormazábal, S Martínez-Canarias, N Perurena, S Vicent & F Lecanda

  2. Bioinformatics and Proteomics Unit, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain

    E Guruceaga

  3. Department of Histology and Pathology, School of Medicine, University of Navarra, Pamplona, Spain

    S Vicent

  4. Bioinformatics and Functional Genomics Research Group, Cancer Research Center, University of Salamanca (CSIC/USAL), Salamanca, Spain

    J De Las Rivas

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Correspondence to F Lecanda.

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Luis-Ravelo, D., Antón, I., Zandueta, C. et al. A gene signature of bone metastatic colonization sensitizes for tumor-induced osteolysis and predicts survival in lung cancer. Oncogene 33, 5090–5099 (2014). https://doi.org/10.1038/onc.2013.440

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