title: A gene signature of bone metastatic colonization sensitizes for tumor-induced osteolysis and predicts survival in lung cancer.
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.
authors.affiliation: Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain.
authors.forename: D
authors.initials: D
authors.lastname: Luis-Ravelo
authors.affiliation: Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain.
authors.forename: I
authors.initials: I
authors.lastname: Antón
authors.affiliation: Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Zandueta
authors.affiliation: Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain.
authors.forename: K
authors.initials: K
authors.lastname: Valencia
authors.affiliation: Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain.
authors.forename: C
authors.initials: C
authors.lastname: Ormazábal
authors.affiliation: Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Martínez-Canarias
authors.affiliation: Bioinformatics and Proteomics Unit, Center for Applied Medical Research (CIMA), University of Navarra, Pamplona, Spain.
authors.forename: E
authors.initials: E
authors.lastname: Guruceaga
authors.affiliation: Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain.
authors.forename: N
authors.initials: N
authors.lastname: Perurena
authors.affiliation: 1] Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain [2] Department of Histology and Pathology, School of Medicine, University of Navarra, Pamplona, Spain.
authors.forename: S
authors.initials: S
authors.lastname: Vicent
authors.affiliation: Bioinformatics and Functional Genomics Research Group, Cancer Research Center, University of Salamanca (CSIC/USAL), Salamanca, Spain.
authors.forename: J
authors.initials: J
authors.lastname: De Las Rivas
authors.affiliation: Division of Oncology, Adhesion and Metastasis Laboratory, University of Navarra, Pamplona, Spain.
authors.forename: F
authors.initials: F
authors.lastname: Lecanda
mesh_terms: D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology; D000077192:Adenocarcinoma of Lung; D000818:Animals; D001859:Bone Neoplasms / Q000235:genetics* / Q000473:pathology / Q000556:secondary*; D045744:Cell Line, Tumor; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D006655:Histone Deacetylases / Q000235:genetics / Q000378:metabolism; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics* / Q000473:pathology; D051379:Mice; D008819:Mice, Nude; D009374:Neoplasms, Experimental; D009419:Nerve Tissue Proteins / Q000235:genetics / Q000378:metabolism*; D010010:Osteoclasts / Q000378:metabolism; D010014:Osteolysis / Q000235:genetics / Q000473:pathology; D051761:Paired Box Transcription Factors / Q000235:genetics / Q000378:metabolism*; D016019:Survival Analysis
chemical_list: D009419:Nerve Tissue Proteins; D051761:Paired Box Transcription Factors; D006655:Histone Deacetylases