The novel kinase inhibitor EMD1214063 is effective against neuroblastoma
- 429 Downloads
Children with high-risk neuroblastoma have poor survival rates, and novel therapies are needed. Previous studies have identified a role for the HGF/c-Met pathway in neuroblastoma pathogenesis. We hypothesized that EMD1214063 would be effective against neuroblastoma tumor cells and tumors in preclinical models via inhibition of HGF/c-Met signaling. Methods We determined the expression of c-Met protein by Western blots in a panel of neuroblastoma tumor cell lines and neuroblastoma cell viability after treatment with EMD1214063 using MTT assays. TUNEL assays and assays for DNA ladder formation, were performed to measure the induction of apoptosis after EMD1214063 treatment. Inhibition of intracellular signaling was measured by Western blot analysis of treated and untreated cells. To investigate the efficacy of EMD1214063 against neuroblastoma tumors in vivo, neuroblastoma cells were injected orthotopically into immunocompromised mice, and mice were treated with oral EMD1214063. Tumors were evaluated for growth, histologic appearance, and induction of apoptosis by immunohistochemistry. Results All neuroblastoma cell lines were sensitive to EMD1214063, and IC50 values ranged from 2.4 to 8.5 μM. EMD1214063 treatment inhibited HGF-mediated c-Met phosphorylation and MEK phosphorylation in neuroblastoma cells. EMD1214063 induced apoptosis in all tested cell lines. In mice with neuroblastoma xenograft tumors, EMD1214063 treatment reduced tumor growth. Conclusions Treatment of neuroblastoma tumor cells with EMD1214063 inhibits HGF-induced c-Met phosphorylation and results in cell death. EMD1214063 treatment is also effective in reducing tumor growth in vivo. EMD1214063 therefore represents a novel therapeutic agent for neuroblastoma, and further preclinical studies of EMD1214063 are warranted.
KeywordsNeuroblastoma EMD1214063 c-Met MEK
Children’s Oncology Group
Terminal deoxynucleotidyl transferase dUTP nick end labeling
American Type Culture Collection
Sodium dodecyl sulfate-polyacrylamide gel electrophoresis
Fetal bovine serum
Hepatocyte growth factor
Nerve growth factor
Dulbecco’s modified Eagle’s medium
3-(4,5 dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide
This study was supported by an Independent Medical Grant from EMD Serono, Inc. to PEZ in the form of research funding and study drug.
Conflict of interest
This study was supported by an Independent Medical Grant from EMD Serono, Inc. to PEZ in the form of research funding and study drug. All other authors declare that they have no conflict of interest.
- 1.Ladenstein R, Philip T, Lasset C, Hartmann O, Zucker JM, Pinkerton R et al (1998) Multivariate analysis of risk factors in stage 4 neuroblastoma patients over the age of one year treated with megatherapy and stem-cell transplantation: a report from the European bone marrow transplantation solid tumor registry. J Clin Oncol 16:953–65PubMedGoogle Scholar
- 2.Matthay KK, Reynolds CP, Seeger RC, Shimada H, Adkins ES, Haas-Kogan D et al (2009) Long-term results for children with high-risk neuroblastoma treated on a randomized trial of myeloablative therapy followed by 13-cis-retinoic acid: a Children’s Oncology Group study. J Clin Oncol 27:1007–13PubMedCentralCrossRefPubMedGoogle Scholar
- 21.Falchook GS, Hong DS, Amin HM, Fu S, Piha-Paul SA, Janku F et al (2013) Phase I study of oral selective c-Met inhibitor EMD1214063 in Pts with advanced solid tumors. J Clin Oncol 31:2506Google Scholar
- 30.Patterson DM, Shohet JM, Kim ES (2011) Preclinical models of pediatric solid tumors (Neuroblastoma) and their use in drug discovery. Curr Protoc Pharmacol 52:14.17.1–14.17.18Google Scholar