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The use of isobaric tag peptide labeling (iTRAQ) and mass spectrometry to examine rare, primitive hematopoietic cells from patients with chronic myeloid leukemia

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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.

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Authors and Affiliations

  1. Division of Cancer Studies, Faculty of Medical and Human Sciences, Christie Hospital, University of Manchester, Wilmslow Road, Manchester, M20 9BX, UK

    Stephen D. Griffiths, Richard D. Unwin & Anthony D. Whetton

  2. Department of Clinical Haematology, Central Manchester and Manchester Children’s University Hospitals NHS Trust, Manchester Royal Infirmary, Oxford Road, Manchester, M13 9WL, UK

    John Burthem & Guy S. Lucas

  3. University Department of Medicine, Royal Infirmary, 10 Alexandra Parade, Glasgow, G31 2ER, Scotland

    Tessa L. Holyoake

  4. Department of Haematology, Imperial College & Hammersmith Hospital, London, W12 0NN, UK

    Junia V. Melo

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  1. Stephen D. Griffiths
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  2. John Burthem
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  3. Richard D. Unwin
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  4. Tessa L. Holyoake
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  5. Junia V. Melo
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Correspondence to Anthony D. Whetton.

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Stephen D. Griffiths and John Burthem contributed equally to this publication. This work is supported by The Leukaemia Research Fund (UK).

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Griffiths, S.D., Burthem, J., Unwin, R.D. et al. The use of isobaric tag peptide labeling (iTRAQ) and mass spectrometry to examine rare, primitive hematopoietic cells from patients with chronic myeloid leukemia. Mol Biotechnol 36, 81–89 (2007). https://doi.org/10.1007/s12033-007-0005-5

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