Flow cytometric determination of Src phosphorylation in pediatric patients treated with dasatinib.
Tyrosine kinase inhibitors, such as imatinib, have dramatically improved the outcomes for patients with selected cancers. For imatinib, western blotting of phospho-CrkL was an insensitive, indirect, and descriptive method to determine drug efficacy. Greater use of targeted therapies should involve more quantitative evaluation of the target's dose-inhibition. The Src/Abl kinase inhibitor dasatinib has recently been approved for use in Ph+ leukemias after failure with imatinib. Src family kinases (SFK) also play a critical role in nonhematologic cancers. We have developed a flow cytometric assay to measure SFK autophosphorylation levels in blood mononuclear cells and observed a direct correlation between its inhibition and patient dosage. This method provides a sensitive, quick, and quantitative tool to assess drug efficacy.
Guerrouahen
Bella S
BS
Division of Pediatrics, MD Anderson Cancer Center, Houston, Texas, USA.
Wieder
Eric
E
Blanchard
Elisabeth G
EG
Lee
Francis Y
FY
Aplenc
Richard
R
Corey
Seth J
SJ
D002648:Child; D000069439:Dasatinib; D005434:Flow Cytometry; D006801:Humans; D008722:Methods; D010766:Phosphorylation; D011743:Pyrimidines / Q000627:therapeutic use*; D013844:Thiazoles / Q000627:therapeutic use*; D019061:src-Family Kinases / Q000378:metabolism*
D011743:Pyrimidines; D013844:Thiazoles; D019061:src-Family Kinases; D000069439:Dasatinib