| Summary: | Despite important advances in the therapy of acute myeloid leukemia
(AML) the majority of patients will die from their disease (Appelbaum,
Rowe, Radich, & Dick, 2001). Characterization of the aberrant molecular
pathways responsible for this malignancy provides a platform to discover
alternative treatments to help alter the fate of patients.
AML is characterized by a blockage in the differentiation of myeloid cells
resulting in the accumulation of highly proliferating immature
hematopoietic cells. Since treatments such as chemotherapy rarely destroy
the leukemic cells entirely, differentiation induction therapy has become a
very attractive treatment option. Interestingly, previous experiments have
shown that ligation of CD44, a cell surface glycoprotein strongly expressed
on all AML cells, with anti-CD44 monoclonal antibodies (mAbs) could
reverse this block in differentiation of leukemic blasts regardless of the AML subtype. To expand the understanding of the cellular regulation and circuitry involved, we aim to apply quantitative phosphoproteomics to monitor dynamic changes in phosphorylation state in response to anti-CD44 treatment.
Protein phosphorylation and dephosphorylation is a highly controlled biochemical process that responds to various intracellular and extracellular stimuli. As phosphorylation is a dynamic process, quantification of these
phosphorylation events would be vastly insightful.
The main objective of this project is to determine the differentiation-dependent
phosphoproteome of AML cells upon treatment of cells with the anti-CD44 mAb.In these experiments, optimization of protein extraction,
phosphopeptide enrichment and data processing and analysis has been achieved. The primary results show successful phosphoproteome extraction complemented with efficient phosphopeptide enrichment and informative data processing. Further quantification with stable isotope
labeling techniques is anticipated to provide candidates for targeted therapy.
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