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The novel kinase inhibitor EMD1214063 is effective against neuroblastoma

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Abstract

Background

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.

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Abbreviations

COG:

Children’s Oncology Group

TUNEL:

Terminal deoxynucleotidyl transferase dUTP nick end labeling

ATCC:

American Type Culture Collection

SDS-PAGE:

Sodium dodecyl sulfate-polyacrylamide gel electrophoresis

7-AAD:

7-amino-actinomycin

FBS:

Fetal bovine serum

HGF:

Hepatocyte growth factor

NGF:

Nerve growth factor

DMEM:

Dulbecco’s modified Eagle’s medium

MTT:

3-(4,5 dimethylthiazolyl-2-yl)-2,5-diphenyltetrazolium bromide

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Grant support

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.

Author information

Authors and Affiliations

  1. Department of Pediatrics, Section of Hematology-Oncology, Baylor College of Medicine, Houston, TX, USA

    Kathy Scorsone, Linna Zhang, Sarah E. Woodfield & Peter E. Zage

  2. Department of Pathology, Baylor College of Medicine, Houston, TX, USA

    John Hicks

  3. Texas Children’s Cancer and Hematology Centers, Baylor College of Medicine, 1102 Bates, Suite 1220, Houston, TX, 77030, USA

    Peter E. Zage

Authors
  1. Kathy Scorsone
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  2. Linna Zhang
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  5. Peter E. Zage
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Corresponding author

Correspondence to Peter E. Zage.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Online Resource 1

Effects of EMD1214063 on neuroblastoma tumor cell morphology. Neuroblastoma tumor cells were treated with increasing concentrations of EMD1214063 for 24 hours, and photographs of cell appearance were obtained. (PDF 34 kb)

Online Resource 2

c-Met expression in neuroblastoma patient tumor samples. Protein lysates from neuroblastoma patient tumors and control cell lines were evaluated for expression of c-Met by Western blot. 3 samples were determined to be positive (#'s 3, 6, and 7). b-actin was used as a control for protein loading. (PDF 29212 kb)

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Scorsone, K., Zhang, L., Woodfield, S.E. et al. The novel kinase inhibitor EMD1214063 is effective against neuroblastoma. Invest New Drugs 32, 815–824 (2014). https://doi.org/10.1007/s10637-014-0107-4

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  • DOI: https://doi.org/10.1007/s10637-014-0107-4

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