The role of necroptosis in acute myeloid leukemia
Acute myeloid leukemia (AML) is the most common form of adult acute leukemia and comprises a phenotypically and genetically heterogeneous group of malignant myeloid neoplasms. In this project, upstream key proteins of the signaling pathways of necroptosis (TNFR1, TNFR2) will be knocked out and their influence on the initiation and persistence of FLT3-ITD-, AML-ETO- and MLL-ENL-induced AML in the mouse model will be investigated. We will use a murine bone marrow transplantation model, which is particularly suitable because the used mutations / translocations were isolated from AML patients and will thus serve as a good model for human leukemias. Furthermore, this model has already been used successfully in several studies In order to characterize the effects of TNFR1 and TNFR2 on initiation and progression of AML, wildtype (WT), TNFR1-, TNFR2-, and TNFR1/2-deficient mice will be treated with 5-fluorouracil (5-FU), which causes an enrichment of the stem cell fraction in the donor bone marrow. Harvested bone marrow cells will be retrovirally transduced with the FLT3-ITD-, AML-ETO- or MLL-ENL-expressing vectors and subsequently transplanted into lethally irradiated WT recipient mice. In a second step we will determine the effects of TNFR1 and TNFR2 on leukemic stem cells via FACS. Therefore, we will be able to establish the method of bone marrow transplantation in Ljubljana and to further evaluate the signaling pathways of necroptosis in AML.