Klin Padiatr 2017; 229(06): 361-366
DOI: 10.1055/s-0037-1607404
Abstracts
Georg Thieme Verlag KG Stuttgart · New York

Development of a novel assay suitable for pre-clinical testing of MAPK inhibitors in low-grade glioma

D Usta
1   Clinical Cooperation Unit Pediatric Oncology, Pre-Clinical Program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
2   German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), Heidelberg, Germany
,
F Selt
1   Clinical Cooperation Unit Pediatric Oncology, Pre-Clinical Program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
2   German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), Heidelberg, Germany
3   Center for Individualized Pediatric Oncology (ZIPO) and Brain Tumors, Translational program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
,
J Hohloch
1   Clinical Cooperation Unit Pediatric Oncology, Pre-Clinical Program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
2   German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), Heidelberg, Germany
,
S Pusch
4   Clinical Cooperation Unit Neuropathology (G380), German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), Heidelberg, Germany
,
SM Pfister
2   German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), Heidelberg, Germany
3   Center for Individualized Pediatric Oncology (ZIPO) and Brain Tumors, Translational program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
5   Division of Pediatric Neurooncology, Pre-Clinical program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
,
DTW Jones
2   German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), Heidelberg, Germany
5   Division of Pediatric Neurooncology, Pre-Clinical program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
,
T Brummer
6   Institute of Molecular Medicine and Cell Research, Albert-Ludwigs-University and University Medical Center, and German Cancer Consortium (DKTK), Freiburg, and German Cancer Research Center (DKFZ), Heidelberg, Germany
,
O Witt
1   Clinical Cooperation Unit Pediatric Oncology, Pre-Clinical Program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
2   German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), Heidelberg, Germany
3   Center for Individualized Pediatric Oncology (ZIPO) and Brain Tumors, Translational program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
,
T Milde
1   Clinical Cooperation Unit Pediatric Oncology, Pre-Clinical Program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
2   German Cancer Research Center (DKFZ) and German Cancer Consortium (DKTK), Heidelberg, Germany
3   Center for Individualized Pediatric Oncology (ZIPO) and Brain Tumors, Translational program, Hopp Children's Cancer Center at the NCT (KiTZ), Heidelberg, Germany
› Author Affiliations
Further Information

Publication History

Publication Date:
25 October 2017 (online)

 

Introduction:

Pilocytic astrocytoma (PA), the most frequent pediatric brain tumor, exhibits an activation of MAPK-signaling by genetic aberration in nearly 100% of the cases, with activating KIAA1549:BRAF-fusions being the most frequent alteration of this pathway (60 – 80%). Surgery, chemotherapy and irradiation can cause significant sequelae without being able to control the disease with certainty. As a result, new therapeutic options with better anti-tumor and fewer side effects are urgently required for the treatment of patients with PA. The aim of this project is to develop an assay suitable for pre-clinical testing of MAPK inhibitors in PA with the ultimate goal to generate pre-clinical data for translational purposes.

Material and methods:

A reporter construct (pDIPZ) was cloned for the use in medium- to high-throughput screening of MAPKi libraries, with a MAPK-responsive ELK1-binding element driving the expression of destabilized GFP (desGFP) and destabilized luciferase (desLuc). Experiments are ongoing to transduce pDIPZ into LGG models (e.g. DKFZ-BT66 with a KIAA1549:BRAF fusion activating the MAPK signaling) using a lentiviral vector. GFP and luciferase activity will be determined by flow cytometry and a luminescence plate reader. MAPKi are used to modulate MAPK activity.

Results:

The pDIPZ reporter construct was successfully packaged in the lentiviral vector. Experiments are ongoing evaluating the dynamic range of the reporter construct's signal and modulation thereof by MAPKi.

Discussion:

After successful generation of a MAPK reporter construct for the use in PA model cells, a full characterization of functional properties will be performed, followed by application of the assay in a MAPKi compound library screening.