The Src and c-Kit kinase inhibitor dasatinib enhances p53-mediated targeting of human acute myeloid leukemia stem cells by chemotherapeutic agents
Authors:
Dos Santos C, McDonald T, Ho YW, Liu H, Lin A, Forman SJ, Kuo YH, Bhatia R.
In:
Source:
Blood
Publication Date:
(
2013
)
Issue:
122(11)
:
1900-13
Research Area:
Immunotherapy / Hematology
Stem Cells
Gene Expression
Basic Research
Experiment
CD34 cells were transfected using the Amaxa 96-well shuttle (Amaxa). 5 3 104 cells were suspended in Nucleofector solution and transfected with siRNA in triplicate, using the human CD34 cell 96-well Nucleofector kit, program E0-100. For p53 siRNA experiments, AML samples transfected with control and p53 siRNA were cultured with or without dasatinib (200 nM) plus DNR (50 nM) for 2 days, and apoptosis was assessed
Abstract
The SRC family kinases (SFKs) and the receptor tyrosine kinase c-Kit are activated in human acute myeloid leukemia (AML) cells. We show here that the SFKs LYN, HCK, or FGR are overexpressed and activated in AML progenitor cells. Treatment with the SFK and c-KIT inhibitor dasatinib selectively inhibits human AML stem/progenitor cell growth in vitro. Importantly, dasatinib markedly increases the elimination of AML stem cells capable of engrafting immunodeficient mice by chemotherapeutic agents. In vivo dasatinib treatment enhances chemotherapy-induced targeting of primary murine AML stem cells capable of regenerating leukemia in secondary recipients. Our studies suggest that enhanced targeting of AML cells by the combination of dasatinib with daunorubicin may be related to inhibition of AKT-mediated human mouse double minute 2 homolog phosphorylation, resulting in enhanced p53 activity in AML cells. Combined treatment using dasatinib and chemotherapy provides a novel approach to increasing p53 activity and enhancing targeting of AML stem cells.
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