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Status |
Public on Nov 22, 2006 |
Title |
Anaplastic Lymphoma Kinase signature |
Organism |
Homo sapiens |
Experiment type |
Expression profiling by array
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Summary |
Anaplastic Large Cell Lymphomas (ALCL) represent a subset of lymphomas in which the Anaplastic Lymphoma Kinase (ALK) gene is frequently fused to the 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/EBPb and the anti-apoptotic 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. Keywords: other
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Overall design |
This series of microarray experiments contains the gene expression profiles of Anaplastic Large Cell Lymphoma (ALCL) cell lines (TS [a subclone of Sup-M2] and Su-DHL1) engineered to express ALK-A5 shRNA under a doxycycline-inducible promoter or treated with cell permeable pyrrolocarbazole-derived ALK inhibitors. A mutated ALK-A5M shRNA was used as control. Briefly, cells were transduced with pLV-DsRed-tTRKRAB, expanded, and used for transduction with pLVTH-GFP-shRNA lentiviral particles. Cells were induced with doxycycline (1 microg/ml) for 12 hours, double GFP+ DsRed+ cells selected by fluorescence-activated cell sorting. Cells expressing GFP in the absence of the inducer were removed by a second flow cytometry sorting, and expanded. shRNA expression was induced by doxycycline treatment for 72 or 84 hours. Drug treatments (300 nM), were performed in TS cells with ALK inhibitors (A2 or A3), mock compound (A1), or control diluent for 6 hours. 5 micrograms of total RNA was processed and hybridized to the Affymetrix HG-U133A chip following the manufacturer's instructions.
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Contributor(s) |
Piva R, Pellegrino E, Mattioli M, Agnelli L, Lombardi L, Boccalatte F, Costa G, Ruggeri BA, Cheng M, Chiarle R, Palestro G, Neri A, Inghirami G |
Citation(s) |
17111047, 25873174 |
Submission date |
Oct 27, 2006 |
Last update date |
May 22, 2019 |
Contact name |
Roberto Piva |
E-mail(s) |
roberto.piva@unito.it
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Phone |
39-011-6336860
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Fax |
39-011-6336887
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Organization name |
University of Torino
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Department |
Department of Pathology and CeRMS
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Lab |
Molecular Oncology
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Street address |
Via Santena 7
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City |
Torino |
ZIP/Postal code |
10126 |
Country |
Italy |
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Platforms (1) |
GPL96 |
[HG-U133A] Affymetrix Human Genome U133A Array |
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Samples (33)
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Relations |
BioProject |
PRJNA100655 |
Supplementary file |
Size |
Download |
File type/resource |
GSE6184_RAW.tar |
113.2 Mb |
(http)(custom) |
TAR (of CEL) |
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