PUBLICATION
A kinome-wide RNAi screen identifies ALK as a target to sensitize neuroblastoma cells for HDAC8-inhibitor treatment
- Authors
- Shen, J., Najafi, S., Stäble, S., Fabian, J., Koeneke, E., Kolbinger, F.R., Wrobel, J.K., Meder, B., Distel, M., Heimburg, T., Sippl, W., Jung, M., Peterziel, H., Kranz, D., Boutros, M., Westermann, F., Witt, O., Oehme, I.
- ID
- ZDB-PUB-180309-4
- Date
- 2018
- Source
- Cell death and differentiation 25(12): 2053-2070 (Journal)
- Registered Authors
- Distel, Martin, Meder, Benjamin
- Keywords
- none
- MeSH Terms
-
- Animals
- Neuroblastoma/drug therapy*
- Neuroblastoma/metabolism
- Neuroblastoma/pathology
- Indoles/pharmacology
- Anaplastic Lymphoma Kinase/antagonists & inhibitors
- Anaplastic Lymphoma Kinase/genetics*
- Anaplastic Lymphoma Kinase/metabolism
- Drug Screening Assays, Antitumor
- Cell Differentiation/drug effects
- Cell Proliferation/drug effects
- Histone Deacetylases/genetics
- Histone Deacetylases/metabolism
- Zebrafish
- Crizotinib/pharmacology
- Protein Kinase Inhibitors/pharmacology*
- Hydroxamic Acids/pharmacology
- Cell Cycle Checkpoints/drug effects
- Humans
- Tumor Cells, Cultured
- RNA Interference*
- Antineoplastic Agents/pharmacology*
- Repressor Proteins/antagonists & inhibitors*
- Repressor Proteins/genetics
- Repressor Proteins/metabolism
- PubMed
- 29515255 Full text @ Cell Death Differ.
Citation
Shen, J., Najafi, S., Stäble, S., Fabian, J., Koeneke, E., Kolbinger, F.R., Wrobel, J.K., Meder, B., Distel, M., Heimburg, T., Sippl, W., Jung, M., Peterziel, H., Kranz, D., Boutros, M., Westermann, F., Witt, O., Oehme, I. (2018) A kinome-wide RNAi screen identifies ALK as a target to sensitize neuroblastoma cells for HDAC8-inhibitor treatment. Cell death and differentiation. 25(12):2053-2070.
Abstract
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.
Genes / Markers
Expression
Phenotype
Mutations / Transgenics
Human Disease / Model
Sequence Targeting Reagents
Fish
Orthology
Engineered Foreign Genes
Mapping