title: Upregulation of MAPK Negative Feedback Regulators and RET in Mutant ALK Neuroblastoma: Implications for Targeted Treatment.
abstract: OBJECTIVE
  Activating ALK mutations are present in almost 10% of primary neuroblastomas and mark patients for treatment with small-molecule ALK inhibitors in clinical trials. However, recent studies have shown that multiple mechanisms drive resistance to these molecular therapies. We anticipated that detailed mapping of the oncogenic ALK-driven signaling in neuroblastoma can aid to identify potential fragile nodes as additional targets for combination therapies.
  
  
  METHODS
  To achieve this goal, transcriptome profiling was performed in neuroblastoma cell lines with the ALK(F1174L) or ALK(R1275Q) hotspot mutations, ALK amplification, or wild-type ALK following pharmacologic inhibition of ALK using four different compounds. Next, we performed cross-species genomic analyses to identify commonly transcriptionally perturbed genes in MYCN/ALK(F1174L) double transgenic versus MYCN transgenic mouse tumors as compared with the mutant ALK-driven transcriptome in human neuroblastomas.
  
  
  RESULTS
  A 77-gene ALK signature was established and successfully validated in primary neuroblastoma samples, in a neuroblastoma cell line with ALK(F1174L) and ALK(R1275Q) regulable overexpression constructs and in other ALKomas. In addition to the previously established PI3K/AKT/mTOR, MAPK/ERK, and MYC/MYCN signaling branches, we identified that mutant ALK drives a strong upregulation of MAPK negative feedback regulators and upregulates RET and RET-driven sympathetic neuronal markers of the cholinergic lineage.
  
  
  CONCLUSIONS
  We provide important novel insights into the transcriptional consequences and the complexity of mutant ALK signaling in this aggressive pediatric tumor. The negative feedback loop of MAPK pathway inhibitors may affect novel ALK inhibition therapies, whereas mutant ALK induced RET signaling can offer novel opportunities for testing ALK-RET oriented molecular combination therapies.
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Irina
authors: I
authors: Lambertz
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Candy
authors: C
authors: Kumps
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Shana
authors: S
authors: Claeys
authors: Department of Pediatric Oncology and Haematology, University Children's Hospital Essen, Germany.
authors: Sven
authors: S
authors: Lindner
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Anneleen
authors: A
authors: Beckers
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Els
authors: E
authors: Janssens
authors: Kids Cancer Center, Sydneys Children's Hospital, Children's Cancer Institute, Lowy Cancer Centre, University of New South Wales, Sydney, Australia.
authors: Daniel R
authors: DR
authors: Carter
authors: Unité Inserm U830, Centre de Recherche, Institut Curie, Paris, France.
authors: Alex
authors: A
authors: Cazes
authors: Kids Cancer Center, Sydneys Children's Hospital, Children's Cancer Institute, Lowy Cancer Centre, University of New South Wales, Sydney, Australia.
authors: Belamy B
authors: BB
authors: Cheung
authors: Biotherapy Unit, IRCCS AOU San Martino-IST, Istituto Nazionale per la Ricerca sul Cancro, Genoa, Italy.
authors: Marilena
authors: M
authors: De Mariano
authors: Oncology Discovery Research and Early Development, Johnson & Johnson, Beerse, Belgium.
authors: An
authors: A
authors: De Bondt
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Sara
authors: S
authors: De Brouwer
authors: Unité Inserm U830, Centre de Recherche, Institut Curie, Paris, France.
authors: Olivier
authors: O
authors: Delattre
authors: VP Oncology, Xcovery LLC, West Palm Beach, Florida.
authors: Jay
authors: J
authors: Gibbons
authors: Unité Inserm U830, Centre de Recherche, Institut Curie, Paris, France.
authors: Isabelle
authors: I
authors: Janoueix-Lerosey
authors: Department of Pediatric Oncology and Haematology, Ghent University Hospital, Ghent, Belgium.
authors: Geneviève
authors: G
authors: Laureys
authors: VP Oncology, Xcovery LLC, West Palm Beach, Florida.
authors: Chris
authors: C
authors: Liang
authors: Kids Cancer Center, Sydneys Children's Hospital, Children's Cancer Institute, Lowy Cancer Centre, University of New South Wales, Sydney, Australia.
authors: Glenn M
authors: GM
authors: Marchall
authors: Center for Human Genetics, K.U. Leuven-VIB, Leuven, Belgium.
authors: Michael
authors: M
authors: Porcu
authors: Department of Pediatrics, Graduate School of Medicine, the University of Tokyo, Tokyo, Japan.
authors: Junko
authors: J
authors: Takita
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: David Camacho
authors: DC
authors: Trujillo
authors: Oncology Discovery Research and Early Development, Johnson & Johnson, Beerse, Belgium.
authors: Ilse
authors: I
authors: Van Den Wyngaert
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Nadine
authors: N
authors: Van Roy
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Alan
authors: A
authors: Van Goethem
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Tom
authors: T
authors: Van Maerken
authors: Center for Human Genetics, K.U. Leuven-VIB, Leuven, Belgium.
authors: Piotr
authors: P
authors: Zabrocki
authors: Center for Human Genetics, K.U. Leuven-VIB, Leuven, Belgium.
authors: Jan
authors: J
authors: Cools
authors: Department of Pediatric Oncology and Haematology, University Children's Hospital Essen, Germany. German Cancer Consortium (DKTK), Germany. Translational Neuro-Oncology, West German Cancer Center, University Hospital Essen, University Duisburg-Essen, Essen, Germany. German Cancer Research Center (DKFZ), Heidelberg, Germany. Centre for Medical Biotechnology, University Duisburg-Essen, Essen, Germany.
authors: Johannes H
authors: JH
authors: Schulte
authors: Oncology Discovery Biology, Janssen Research & Development, a division of Janssen Pharmaceutica NV, Beerse, Belgium.
authors: Jorge
authors: J
authors: Vialard
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium. franki.speleman@ugent.be.
authors: Frank
authors: F
authors: Speleman
authors: Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium.
authors: Katleen
authors: K
authors: De Preter
mesh_terms: D000469:Alkaline Phosphatase / Q000235:genetics*; D000818:Animals; D015153:Blotting, Western; D045744:Cell Line, Tumor; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D025461:Feedback, Physiological; D006801:Humans; D051379:Mice; D008822:Mice, Transgenic; D020929:Mitogen-Activated Protein Kinase Kinases / Q000378:metabolism*; D058990:Molecular Targeted Therapy / Q000379:methods*; D009447:Neuroblastoma / Q000235:genetics* / Q000378:metabolism; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D051096:Proto-Oncogene Proteins c-ret / Q000378:metabolism*; D059467:Transcriptome; D015854:Up-Regulation
chemical_list: D051096:Proto-Oncogene Proteins c-ret; D020929:Mitogen-Activated Protein Kinase Kinases; D000469:Alkaline Phosphatase