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Investigational New Drugs

, Volume 32, Issue 5, pp 815–824 | Cite as

The novel kinase inhibitor EMD1214063 is effective against neuroblastoma

  • Kathy Scorsone
  • Linna Zhang
  • Sarah E. Woodfield
  • John Hicks
  • Peter E. ZageEmail author
PRECLINICAL STUDIES

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.

Keywords

Neuroblastoma EMD1214063 c-Met MEK 

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

Notes

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.

Supplementary material

10637_2014_107_MOESM1_ESM.pdf (35 kb)
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)
10637_2014_107_MOESM2_ESM.pdf (28.5 mb)
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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Copyright information

© Springer Science+Business Media New York 2014

Authors and Affiliations

  • Kathy Scorsone
    • 1
  • Linna Zhang
    • 1
  • Sarah E. Woodfield
    • 1
  • John Hicks
    • 2
  • Peter E. Zage
    • 1
    • 3
    Email author
  1. 1.Department of Pediatrics, Section of Hematology-OncologyBaylor College of MedicineHoustonUSA
  2. 2.Department of PathologyBaylor College of MedicineHoustonUSA
  3. 3.Texas Children’s Cancer and Hematology CentersBaylor College of MedicineHoustonUSA

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