Autophagy in normal hematopoiesis and leukemia

Here we have explored whether inhibition of autophagy can be used as treatment strategy for acute myeloid leukemia (AML). Steady-state autophagy was measured in leukemic cell lines and primary human CD34 + AML cells with a large variability in basal autophagy between AMLs observed. The autophagy-flux was higher in AMLs classified as poor-risk, which are frequently associated with TP53 mutations (TP53 mut ), compared to favorable- and intermediate-risk AMLs. In addition, the higher flux was associated with a higher expression level of several autophagy genes, but was not affected by alterations in p53 expression by knocking down p53 or overexpression of wild type p53 or p53 R273H . AML CD34 + cells were more sensitive to the autophagy inhibitor hydroxychloroquine (HCQ) than normal bone marrow CD34 + cells. Similar, inhibition of autophagy by knockdown of ATG5 or ATG7 triggered apoptosis, which coincided with increased expression of p53. In contrast to wild type p53 AML (TP53 wt ), HCQ treatment did not trigger a BAX and PUMA-dependent apoptotic response in AMLs harboring TP53 mut . To further characterize autophagy in the leukemic stem cell (LSC)-enriched cell fraction AML CD34 + cells were separated into ROS low and ROS high subfractions. The immature AML-CD34 + -enriched ROS low cells maintained higher basal autophagy and showed reduced survival upon HCQ treatment compared to ROS high cells. Finally, knockdown of ATG5 inhibits in vivo maintenance of AML CD34 + cells in NSG mice. These results indicate that targeting autophagy might provide new therapeutic options for treatment of AML since it affects the immature AML subfraction. cells on these effector pathways. The aim of our study was to determine whether inhibiting autophagy can provide an additional means to impair LSC functionality. We demonstrated that AML CD34 + cells are more susceptible for autophagy inhibition than normal CD34 + cells. P53 is an important effector pathway in the observed apoptotic responds, triggered by inhibition of autophagy.

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