The small molecule inhibitor YK-4-279 disrupts mitotic progression of neuroblastoma cells, overcomes drug resistance and synergizes with inhibitors of mitosis

• Published in: Cancer letters 2017-09, Vol.403, p.74-85
• Main Authors: Kollareddy, Madhu, Sherrard, Alice, Park, Ji Hyun, Szemes, Marianna, Gallacher, Kelli, Melegh, Zsombor, Oltean, Sebastian, Michaelis, Martin, Cinatl, Jindrich, Kaidi, Abderrahmane, Malik, Karim
• Format: Article
• Language: English
• Subjects: Acetylation; Antimitotic Agents - pharmacology; Antineoplastic Combined Chemotherapy Protocols - pharmacology; Apoptosis; Apoptosis - drug effects; Aurora kinase; Aurora Kinase A - antagonists & inhibitors; Aurora Kinase A - metabolism; Azepines - pharmacology; Cancer therapies; Cell culture; Cell cycle; Cell Cycle - drug effects; Cell death; Cell division; Cell Line, Tumor; Chemical compounds; Chemotherapy; Dose-Response Relationship, Drug; Drug dosages; Drug resistance; Drug Resistance, Multiple; Drug Resistance, Neoplasm; Drug resistance/synergy; Drug Synergism; Enzyme inhibitors; Extracellular signal-regulated kinase; Flow cytometry; Gene amplification; Gene expression; Genomes; Hematology, Oncology and Palliative Medicine; Humans; Indoles - pharmacology; Inhibitors; Inhibitory Concentration 50; Kinases; Kinetochores - drug effects; Kinetochores - pathology; Malignancy; Metabolic pathways; Microtubules; Mitosis; Mitosis - drug effects; Molecules; Mutation; Neuroblastoma; Neuroblastoma - drug therapy; Neuroblastoma - genetics; Neuroblastoma - metabolism; Neuroblastoma - pathology; Neuroblasts; Original; Paclitaxel; Paclitaxel - pharmacology; Poisons; Prometaphase - drug effects; Protein Kinase Inhibitors - pharmacology; Proteins; Pyrimidines - pharmacology; RNA Interference; Signal Transduction - drug effects; Spindle Apparatus - drug effects; Spindle Apparatus - pathology; Spindles; Time Factors; Transcription factors; Transfection; Tumor Suppressor Protein p53 - genetics; Tumor Suppressor Protein p53 - metabolism; Vincristine; Vincristine - pharmacology; YK-4-279
• ISSN: 0304-3835; 1872-7980
• DOI: 10.1016/j.canlet.2017.05.027

Abstract Neuroblastoma is a biologically and clinically heterogeneous pediatric malignancy that includes a high-risk subset for which new therapeutic agents are urgently required. As well as MYCN amplification, activating point mutations of ALK and NRAS are associated with high-risk and relapsing neuroblastoma. As both ALK and RAS signal through the MEK/ERK pathway, we sought to evaluate two previously reported inhibitors of ETS-related transcription factors, which are transcriptional mediators of the Ras-MEK/ERK pathway in other cancers. Here we show that YK-4-279 suppressed growth and triggered apoptosis in nine neuroblastoma cell lines, while BRD42048, another ETV1 inhibitor, was ineffective. These results suggest that YK-4-279 acts independently of ETS-related transcription factors. Further analysis reveals that YK-4-279 induces mitotic arrest in prometaphase, resulting in subsequent cell death. Mechanistically, we show that YK-4-279 inhibits the formation of kinetochore microtubules, with treated cells showing a broad range of abnormalities including multipolar, fragmented and unseparated spindles, together leading to disrupted progression through mitosis. Notably, YK-4-279 does not affect microtubule acetylation, unlike the conventional mitotic poisons paclitaxel and vincristine. Consistent with this, we demonstrate that YK-4-279 overcomes vincristine-induced resistance in two neuroblastoma cell-line models. Furthermore, combinations of YK-4-279 with vincristine, paclitaxel or the Aurora kinase A inhibitor MLN8237/Alisertib show strong synergy, particularly at low doses. Thus, YK-4-279 could potentially be used as a single-agent or in combination therapies for the treatment of high-risk and relapsing neuroblastoma, as well as other cancers.