Kinome profiling in pediatric brain tumors as a new approach for target discovery.
Progression in pediatric brain tumor growth is thought to be the net result of signaling through various protein kinase-mediated networks driving cell proliferation. Defining new targets for treatment of human malignancies, without a priori knowledge on aberrant cell signaling activity, remains exceedingly complicated. Here, we introduce kinome profiling using flow-through peptide microarrays as a new concept for target discovery. Comprehensive tyrosine kinase activity profiles were identified in 29 pediatric brain tumors using the PamChip kinome profiling system. Previously reported activity of epidermal growth factor receptor, c-Met, and vascular endothelial growth factor receptor in pediatric brain tumors could be appreciated in our array results. Peptides corresponding with phosphorylation consensus sequences for Src family kinases showed remarkably high levels of phosphorylation compared with normal tissue types. Src activity was confirmed applying Phos-Tag SDS-PAGE. Furthermore, the Src family kinase inhibitors PP1 and dasatinib induced substantial tumor cell death in nine pediatric brain tumor cell lines but not in control cell lines. Thus, this study describes a new high-throughput technique to generate clinically relevant tyrosine kinase activity profiles as has been shown here for pediatric brain tumors. In the era of a rapidly increasing number of small-molecule inhibitors, this approach will enable us to rapidly identify new potential targets in a broad range of human malignancies.
Sikkema
Arend H
AH
Department of Pediatric Oncology, University Medical Center Groningen, University of Groningen, Groningen, The Netherlands.
Diks
Sander H
SH
den Dunnen
Wilfred F A
WF
ter Elst
Arja
A
Scherpen
Frank J G
FJ
Hoving
Eelco W
EW
Ruijtenbeek
Rob
R
Boender
Piet J
PJ
de Wijn
Rik
R
Kamps
Willem A
WA
Peppelenbosch
Maikel P
MP
de Bont
Eveline S J M
ES
D001932:Brain Neoplasms / Q000201:enzymology* / Q000473:pathology; D002460:Cell Line; D045744:Cell Line, Tumor; D049109:Cell Proliferation; D002470:Cell Survival; D002648:Child; D016000:Cluster Analysis; D018922:HL-60 Cells; D006801:Humans; D015151:Immunoblotting; D020014:K562 Cells; D046228:Microarray Analysis / Q000379:methods*; D010455:Peptides / Q000145:classification / Q000378:metabolism; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000378:metabolism*; D015203:Reproducibility of Results; D019061:src-Family Kinases / Q000378:metabolism
D010455:Peptides; D011505:Protein-Tyrosine Kinases; D019061:src-Family Kinases