title: The use of isobaric tag peptide labeling (iTRAQ) and mass spectrometry to examine rare, primitive hematopoietic cells from patients with chronic myeloid leukemia.
abstract: Chronic Myeloid Leukemia (CML) is a hematopoietic stem cell disease, associated with a t(9, 22) chromosomal translocation leading to formation of the BCR/ABL chimeric protein, which has an intrinsic tyrosine kinase activity. Recently, the BCR/ABL tyrosine kinase inhibitor imatinib mesylate (imatinib) has been successfully used clinically, although, disease relapse can still occur. The precise detail of the mechanism by which CML cells respond to imatinib is still unclear. We therefore systematically examined the effects of imatinib on the primitive CML cell proteome, having first established that the drug inhibits proliferation and induces increased apoptosis and differentiation. To define imatinib-induced effects on the CML proteome, we employed isobaric tag peptide labeling (iTRAQ) coupled to two-dimensional liquid chromatography/tandem mass spectrometry. Given the limited clinical material available, the isobaric tag approach identified a large population of proteins and provided relative quantification on four samples at once. Novel consequences of the action of imatinib were identified using this mass spectrometric approach. DEAD-box protein 3, heat shock protein 105 kDa, and peroxiredoxin-3 were identified as potential protein markers for response to imatinib.
authors.affiliation: Division of Cancer Studies, Faculty of Medical and Human Sciences, Christie Hospital, University of Manchester, Wilmslow Road, Manchester M20 9BX, UK.
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Griffiths
authors.forename: John
authors.initials: J
authors.lastname: Burthem
authors.forename: Richard D
authors.initials: RD
authors.lastname: Unwin
authors.forename: Tessa L
authors.initials: TL
authors.lastname: Holyoake
authors.forename: Junia V
authors.initials: JV
authors.lastname: Melo
authors.forename: Guy S
authors.initials: GS
authors.lastname: Lucas
authors.forename: Anthony D
authors.initials: AD
authors.lastname: Whetton
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D017209:Apoptosis; D001549:Benzamides; D002454:Cell Differentiation / Q000187:drug effects; D049109:Cell Proliferation / Q000187:drug effects; D002853:Chromatography, Liquid / Q000379:methods; D053487:DEAD-box RNA Helicases / Q000032:analysis; D050957:HSP110 Heat-Shock Proteins / Q000032:analysis; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D000068877:Imatinib Mesylate; D015464:Leukemia, Myelogenous, Chronic, BCR-ABL Positive / Q000378:metabolism*; D010455:Peptides / Q000737:chemistry; D010544:Peroxidases / Q000032:analysis; D061105:Peroxiredoxin III; D054464:Peroxiredoxins; D010879:Piperazines / Q000494:pharmacology*; D047428:Protein Kinase Inhibitors / Q000494:pharmacology*; D020543:Proteome / Q000032:analysis / Q000187:drug effects*; D040901:Proteomics / Q000379:methods*; D011743:Pyrimidines / Q000494:pharmacology*; D053719:Tandem Mass Spectrometry / Q000379:methods; D014407:Tumor Cells, Cultured
chemical_list: D000970:Antineoplastic Agents; D001549:Benzamides; D050957:HSP110 Heat-Shock Proteins; C097205:HSPH1 protein, human; D010455:Peptides; D010879:Piperazines; D047428:Protein Kinase Inhibitors; D020543:Proteome; D011743:Pyrimidines; D000068877:Imatinib Mesylate; D010544:Peroxidases; C516115:PRDX3 protein, human; D061105:Peroxiredoxin III; D054464:Peroxiredoxins; C119332:DDX3X protein, human; D053487:DEAD-box RNA Helicases