title: A phosphoproteomics approach to identify candidate kinase inhibitor pathway targets in lymphoma-like primary cell lines.
abstract: Mass spectrometry-based technologies are increasingly utilized in drug discovery. Phosphoproteomics in particular has allowed for the efficient surveying of phosphotyrosine signaling pathways involved in various diseases states, most prominently in cancer. We describe a phosphotyrosine-based proteomics screening approach to identify signaling pathways and tyrosine kinase inhibitor targets in highly tumorigenic human lymphoma-like primary cells. We identified several receptor tyrosine kinase pathways and validated SRC family kinases (SFKs) as potential drug targets for targeted selection of small molecule inhibitors. BMS-354825 (dasatinib) and SKI-606 (bosutinib), second and third generation clinical SFK/ABL inhibitors, were found to be potent cytotoxic agents against tumorigenic cells with low toxicity to normal pediatric stem cells. Both SFK inhibitors reduced ERK1/2 and AKT phosphorylation and induced apoptosis. This study supports the adaptation of high-end mass spectrometry techniques for the efficient identification of candidate tyrosine kinases as novel therapeutic targets in primary cancer cell lines.
authors.affiliation: Cell Biology Program, The Hospital for Sick Children, TMDT East Tower #12-314, 101 College St., Toronto, Ontario, M5G 1L7; Canada. dkaplan@sickkids.ca.
authors.forename: Miliana
authors.initials: M
authors.lastname: Vojvodic
authors.forename: Loen M
authors.initials: LM
authors.lastname: Hansford
authors.forename: Olena
authors.initials: O
authors.lastname: Morozova
authors.forename: Kim M
authors.initials: KM
authors.lastname: Blakely
authors.forename: Paul
authors.initials: P
authors.lastname: Taylor
authors.forename: Kelly E
authors.initials: KE
authors.lastname: Fathers
authors.forename: Jason
authors.initials: J
authors.lastname: Moffat
authors.forename: Marco
authors.initials: M
authors.lastname: Marra
authors.forename: Kristen M
authors.initials: KM
authors.lastname: Smith
authors.forename: Michael F
authors.initials: MF
authors.lastname: Moran
authors.forename: David R
authors.initials: DR
authors.lastname: Kaplan
mesh_terms: D000814:Aniline Compounds / Q000494:pharmacology*; D017209:Apoptosis / Q000187:drug effects; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D000069439:Dasatinib; D020869:Gene Expression Profiling; D006801:Humans; D008223:Lymphoma; D019950:Mitogen-Activated Protein Kinase 1 / Q000378:metabolism; D048052:Mitogen-Activated Protein Kinase 3 / Q000378:metabolism; D009570:Nitriles / Q000494:pharmacology*; D010766:Phosphorylation; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D040901:Proteomics; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism; D011743:Pyrimidines / Q000494:pharmacology*; D011804:Quinolines / Q000494:pharmacology*; D013844:Thiazoles / Q000494:pharmacology*; D019061:src-Family Kinases / Q000235:genetics*
chemical_list: D000814:Aniline Compounds; D009570:Nitriles; D047428:Protein Kinase Inhibitors; D011743:Pyrimidines; D011804:Quinolines; D013844:Thiazoles; C471992:bosutinib; D019061:src-Family Kinases; D051057:Proto-Oncogene Proteins c-akt; D019950:Mitogen-Activated Protein Kinase 1; D048052:Mitogen-Activated Protein Kinase 3; D000069439:Dasatinib