Bead-based profiling of tyrosine kinase phosphorylation identifies SRC as a potential target for glioblastoma therapy.
The aberrant activation of tyrosine kinases represents an important oncogenic mechanism, and yet the majority of such events remain undiscovered. Here we describe a bead-based method for detecting phosphorylation of both wild-type and mutant tyrosine kinases in a multiplexed, high-throughput and low-cost manner. With the aim of establishing a tyrosine kinase-activation catalog, we used this method to profile 130 human cancer lines. Follow-up experiments on the finding that SRC is frequently phosphorylated in glioblastoma cell lines showed that SRC is also activated in primary glioblastoma patient samples and that the SRC inhibitor dasatinib (Sprycel) inhibits viability and cell migration in vitro and tumor growth in vivo. Testing of dasatinib-resistant tyrosine kinase alleles confirmed that SRC is indeed the relevant target of dasatinib, which inhibits many tyrosine kinases. These studies establish the feasibility of tyrosine kinome-wide phosphorylation profiling and point to SRC as a possible therapeutic target in glioblastoma.
Du
Jinyan
J
The Broad Institute of Harvard University and Massachusetts Institute of Technology, 7 Cambridge Center, Cambridge, Massachusetts 02142, USA.
Bernasconi
Paula
P
Clauser
Karl R
KR
Mani
D R
DR
Finn
Stephen P
SP
Beroukhim
Rameen
R
Burns
Melissa
M
Julian
Bina
B
Peng
Xiao P
XP
Hieronymus
Haley
H
Maglathlin
Rebecca L
RL
Lewis
Timothy A
TA
Liau
Linda M
LM
Nghiemphu
Phioanh
P
Mellinghoff
Ingo K
IK
Louis
David N
DN
Loda
Massimo
M
Carr
Steven A
SA
Kung
Andrew L
AL
Golub
Todd R
TR
D000818:Animals; D001709:Biotechnology / Q000379:methods*; D045744:Cell Line, Tumor; D000069439:Dasatinib; D019008:Drug Resistance, Neoplasm; D005821:Genetic Techniques; D005909:Glioblastoma / Q000201:enzymology / Q000628:therapy*; D006801:Humans; D008297:Male; D051379:Mice; D008819:Mice, Nude; D009368:Neoplasm Transplantation; D010766:Phosphorylation; D011505:Protein-Tyrosine Kinases / Q000737:chemistry*; D011743:Pyrimidines / Q000494:pharmacology; D013844:Thiazoles / Q000494:pharmacology; D019061:src-Family Kinases / Q000378:metabolism
D011743:Pyrimidines; D013844:Thiazoles; D011505:Protein-Tyrosine Kinases; D019061:src-Family Kinases; D000069439:Dasatinib