Abstract
Targeting of leukemic stem cells with specific immunotherapy would be an ideal approach for the treatment of myeloid malignancies, but suitable epitopes are unknown. The comparative proteome-level characterization of hematopoietic stem and progenitor cells from healthy stem cell donors and patients with acute myeloid leukemia has the potential to reveal differentially expressed proteins which can be used as surface-markers or as proxies for affected molecular pathways. We employed mass spectrometry methods to analyze the proteome of the cytosolic and the membrane fraction of CD34 and CD123 co-expressing FACS-sorted leukemic progenitors from five patients with acute myeloid leukemia. As a reference, CD34+CD123+ normal hematopoietic progenitor cells from five healthy, granulocyte-colony stimulating factor (G-CSF) mobilized stem cell donors were analyzed. In this Tandem Mass Tag (TMT) 10-plex labelling–based approach, 2070 proteins were identified with 171 proteins differentially abundant in one or both cellular compartments. This proof-of-principle-study demonstrates the potential of mass spectrometry to detect differentially expressed proteins in two compartment fractions of the entire proteome of leukemic stem cells, compared to their non-malignant counterparts. This may contribute to future immunotherapeutic target discoveries and individualized AML patient characterization.
Highlights
During the past 15 years, mass spectrometry has undergone significant technological improvements allowing the rapid identification of large numbers of proteins today
Acute myeloid leukemia stem cells are the most important target cells in immunotherapeutic approaches aimed at the growth) are the most important target cells in immunotherapeutic approaches aimed at the eradication eradication of Acute Myeloid Leukemia (AML) cells
Limited data is available on the proteome of those cells, which is needed of AML cells
Summary
During the past 15 years, mass spectrometry has undergone significant technological improvements allowing the rapid identification of large numbers of proteins today. In the search for leukemia specific proteins, by applying two-dimensional electrophoresis combined with mass spectrometry methods, Ota et al identified a limited average total number of 168 proteins in 13 samples of CD133+ hematopoietic stem cell-like cells from patients with various hematologic malignancies, including five AML samples. They identified 11 differentially expressed protein forms in at least one of the 13 leukemic samples [1]. Luczak et al analyzed leukemic cells from 38 patients with AML subtypes M1 and M2 and 17 healthy volunteers using 2D gel electrophoresis and mass spectrometry In their comparative analysis, they identified 25 differentially regulated proteins, among which they found five potential AML biomarkers. The method described in this pilot study may contribute to the establishment of an attractive diagnostic tool which might be used to validate candidate target molecules expressed by LSCs for personalized therapeutic strategies
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