Abstract B4: Growth-factor-driven rescue to receptor tyrosine kinase inhibitors in pediatric low-grade astrocytoma and ependymoma
Authors: Sie Mariska , Dunnen Wilfred F.A. den , Lourens Harm Jan , Boer Tiny G.J. Meeuwsen-de , Scherpen Frank J.G. , Kampen Kim R. , Hoving Eelco W. , Bont Eveline S.J.M. de 
Published: May 2015

Introduction: Brain tumors are still the leading cause of cancer morbidity and mortality among children, despite different therapeutic options including neurosurgery, chemotherapy and radiation. Up to now, several clinical studies have been started investigating the importance of receptor tyrosine kinase (RTK) inhibitors upon progression free survival in various pediatric brain tumors. However, it has been shown that single targeted kinase inhibition failed due to tumor resistance mechanisms. The present study will extend our previous observations that vascular endothelial growth receptor (VEGFR) 2, platelet derived growth factor receptor (PDGFR) β, Src, the epidermal growth factor receptor (ErbB) family, and hepatocyte growth factor receptor (HGFR/cMet) are potential drugable targets in pediatric low grade astrocytoma and ependymoma with investigations concerning growth-factor-driven rescue.

Material and methods: Expression of various RTKs in pediatric low grade astrocytoma (Res-186, Res-259, UW-467) and ependymoma (Res-196) cell lines were determined using flow cytometry analyses. Growth-factor-driven rescue during RTK inhibition with agents including sorafenib, dasatinib, canertinib and crizotinib were analyzed on cell viability level measured with WST-1 cell viability assays and on phosphorylation level of the crucial downstream PI3K/Akt and MAPK/Erk survival signaling pathways using western blot analyses.

Results: VEGFR-1, fibroblast growth factor receptor (FGFR) 1, ErbB-1, HGFR and recepteur d'origine nantais (RON) were highly expressed (respectively 52-77%, 34-51%, 63-90%, 83-98%, 65-95%) and their respective inhibitors showed induced decrease of tumor cell viability. EGF, HGF and FGF, which are normally environmentally expressed in brain (tumor) tissue, showed to be effective rescue inducing growth factors resulting in increased cell survival especially during treatment with dasatinib (complete rescue) or sorafenib (partial rescue) in both low grade astrocytoma and ependymoma. Growth-factor-driven rescue was less prominent when canertinib or crizotinib were used. Rescue was underscored by activating downstream Akt and/or Erk phosphorylation. Combination treatment showed to be able to overcome the growth-factor-driven resistance.

Conclusions: In conclusion, our study highlights the extensive importance of environmentally present growth factors in developing resistance towards RTK inhibitors. It is of great interest to anticipate upon these results for the design of new therapeutic trials with RTK inhibitors in pediatric low grade astrocytoma and ependymoma.

Citation Format: Mariska Sie, Wilfred F.A. den Dunnen, Harm Jan Lourens, Tiny G.J. Meeuwsen-de Boer, Frank J.G. Scherpen, Kim R. Kampen, Eelco W. Hoving, Eveline S.J.M. de Bont. Growth-factor-driven rescue to receptor tyrosine kinase inhibitors in pediatric low-grade astrocytoma and ependymoma. [abstract]. In: Proceedings of the AACR Special Conference on Pediatric Cancer at the Crossroads: Translating Discovery into Improved Outcomes; Nov 3-6, 2013; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2013;74(20 Suppl):Abstract nr B4.