A kinome-wide RNAi screen identifies ALK as a target to sensitize neuroblastoma cells for HDAC8-inhibitor treatment
| Autor: | E Koeneke, Olaf Witt, Sara Najafi, Frank Westermann, Benjamin Meder, Manfred Jung, Fiona R. Kolbinger, Martin Distel, Johannes Fabian, Ina Oehme, Sina Stäble, Dominique Kranz, Tino Heimburg, Heike Peterziel, Wolfgang Sippl, Jagoda K. Wrobel, Michael Boutros, Jing Shen |
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| Rok vydání: | 2018 |
| Předmět: |
0301 basic medicine
Indoles Antineoplastic Agents Hydroxamic Acids Article Histone Deacetylases Receptor tyrosine kinase Neuroblastoma 03 medical and health sciences Crizotinib RNA interference Differentiation therapy hemic and lymphatic diseases Tumor Cells Cultured medicine Animals Humans Anaplastic Lymphoma Kinase Kinome Protein Kinase Inhibitors Molecular Biology Zebrafish Cell Proliferation biology Cell Differentiation HDAC8 Cell Cycle Checkpoints Cell Biology Cell cycle medicine.disease Repressor Proteins 030104 developmental biology biology.protein Cancer research RNA Interference Drug Screening Assays Antitumor medicine.drug |
| Zdroj: | Cell Death and Differentiation |
| ISSN: | 1476-5403 1350-9047 |
| Popis: | The prognosis of advanced stage neuroblastoma patients remains poor and, despite intensive therapy, the 5-year survival rate remains less than 50%. We previously identified histone deacetylase (HDAC) 8 as an indicator of poor clinical outcome and a selective drug target for differentiation therapy in vitro and in vivo. Here, we performed kinome-wide RNAi screening to identify genes that are synthetically lethal with HDAC8 inhibitors. These experiments identified the neuroblastoma predisposition gene ALK as a candidate gene. Accordingly, the combination of the ALK/MET inhibitor crizotinib and selective HDAC8 inhibitors (3-6 µM PCI-34051 or 10 µM 20a) efficiently killed neuroblastoma cell lines carrying wildtype ALK (SK-N-BE(2)-C, IMR5/75), amplified ALK (NB-1), and those carrying the activating ALK F1174L mutation (Kelly), and, in cells carrying the activating R1275Q mutation (LAN-5), combination treatment decreased viable cell count. The effective dose of crizotinib in neuroblastoma cell lines ranged from 0.05 µM (ALK-amplified) to 0.8 µM (wildtype ALK). The combinatorial inhibition of ALK and HDAC8 also decreased tumor growth in an in vivo zebrafish xenograft model. Bioinformatic analyses revealed that the mRNA expression level of HDAC8 was significantly correlated with that of ALK in two independent patient cohorts, the Academic Medical Center cohort (n = 88) and the German Neuroblastoma Trial cohort (n = 649), and co-expression of both target genes identified patients with very poor outcome. Mechanistically, HDAC8 and ALK converge at the level of receptor tyrosine kinase (RTK) signaling and their downstream survival pathways, such as ERK signaling. Combination treatment of HDAC8 inhibitor with crizotinib efficiently blocked the activation of growth receptor survival signaling and shifted the cell cycle arrest and differentiation phenotype toward effective cell death of neuroblastoma cell lines, including sensitization of resistant models, but not of normal cells. These findings reveal combined targeting of ALK and HDAC8 as a novel strategy for the treatment of neuroblastoma. |
| Databáze: | OpenAIRE |
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