title: Functional validation of the anaplastic lymphoma kinase signature identifies CEBPB and BCL2A1 as critical target genes.
abstract: Anaplastic large cell lymphomas (ALCLs) represent a subset of lymphomas in which the anaplastic lymphoma kinase (ALK) gene is frequently fused to the nucleophosmin (NPM) gene. We previously demonstrated that the constitutive phosphorylation of ALK chimeric proteins is sufficient to induce cellular transformation in vitro and in vivo and that ALK activity is strictly required for the survival of ALK-positive ALCL cells. To elucidate the signaling pathways required for ALK-mediated transformation and tumor maintenance, we analyzed the transcriptomes of multiple ALK-positive ALCL cell lines, abrogating their ALK-mediated signaling by inducible ALK RNA interference (RNAi) or with potent and cell-permeable ALK inhibitors. Transcripts derived from the gene expression profiling (GEP) analysis uncovered a reproducible signature, which included a novel group of ALK-regulated genes. Functional RNAi screening on a set of these ALK transcriptional targets revealed that the transcription factor C/EBPbeta and the antiapoptotic protein BCL2A1 are absolutely necessary to induce cell transformation and/or to sustain the growth and survival of ALK-positive ALCL cells. Thus, we proved that an experimentally controlled and functionally validated GEP analysis represents a powerful tool to identify novel pathogenetic networks and validate biologically suitable target genes for therapeutic interventions.
authors.affiliation: Department of Pathology and Center for Experimental Research and Medical Studies (CeRMS), University of Turin, Turin, Italy. roberto.piva@unito.it
authors.forename: Roberto
authors.initials: R
authors.lastname: Piva
authors.forename: Elisa
authors.initials: E
authors.lastname: Pellegrino
authors.forename: Michela
authors.initials: M
authors.lastname: Mattioli
authors.forename: Luca
authors.initials: L
authors.lastname: Agnelli
authors.forename: Luigia
authors.initials: L
authors.lastname: Lombardi
authors.forename: Francesco
authors.initials: F
authors.lastname: Boccalatte
authors.forename: Giulia
authors.initials: G
authors.lastname: Costa
authors.forename: Bruce A
authors.initials: BA
authors.lastname: Ruggeri
authors.forename: Mangeng
authors.initials: M
authors.lastname: Cheng
authors.forename: Roberto
authors.initials: R
authors.lastname: Chiarle
authors.forename: Giorgio
authors.initials: G
authors.lastname: Palestro
authors.forename: Antonino
authors.initials: A
authors.lastname: Neri
authors.forename: Giorgio
authors.initials: G
authors.lastname: Inghirami
mesh_terms: D000077548:Anaplastic Lymphoma Kinase; D000818:Animals; D017209:Apoptosis / Q000235:genetics; D022782:CCAAT-Enhancer-Binding Protein-beta / Q000235:genetics* / Q000378:metabolism; D002453:Cell Cycle / Q000235:genetics; D002460:Cell Line; D002470:Cell Survival / Q000235:genetics; D002471:Cell Transformation, Neoplastic / Q000235:genetics; D005434:Flow Cytometry; D020869:Gene Expression Profiling / Q000379:methods; D006801:Humans; D016403:Lymphoma, Large B-Cell, Diffuse / Q000235:genetics / Q000503:physiopathology; D051379:Mice; D015778:Minor Histocompatibility Antigens; D009369:Neoplasms / Q000235:genetics / Q000378:metabolism; D009687:Nuclear Proteins / Q000235:genetics / Q000378:metabolism; D000090243:Nucleophosmin; D020411:Oligonucleotide Array Sequence Analysis; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000037:antagonists & inhibitors / Q000235:genetics* / Q000378:metabolism; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics* / Q000378:metabolism; D034622:RNA Interference; D020794:Receptor Protein-Tyrosine Kinases; D020133:Reverse Transcriptase Polymerase Chain Reaction; D050796:STAT3 Transcription Factor / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction / Q000235:genetics / Q000502:physiology
chemical_list: C096816:BCL2-related protein A1; D022782:CCAAT-Enhancer-Binding Protein-beta; D015778:Minor Histocompatibility Antigens; C000716967:NPM1 protein, human; D009687:Nuclear Proteins; D019253:Proto-Oncogene Proteins c-bcl-2; D050796:STAT3 Transcription Factor; D000090243:Nucleophosmin; C000626173:ALK protein, human; C000626174:Alk protein, mouse; D000077548:Anaplastic Lymphoma Kinase; D011505:Protein-Tyrosine Kinases; D020794:Receptor Protein-Tyrosine Kinases