-Catenin contributes to leukemogenesis induced by AML-associated translocation products by increasing the self-renewal of very primitive progenitor cells

The t(6;9) associated DEK/CAN fusion protein targets a population of long-term repopulating hematopoietic stem cells for leukemogenic transformation

“…The ectopic expression of DEK/CAN used a retroviral transduction system on the basis of the PINCO vector as previously described (Supplementary Figure 1a). 5 The efficient translation and integrity of the DEK/CAN open reading frame was confirmed by in vitro translation (data not shown) and by expression in several cell lines, as well as in Sca1 þ /lin À primary murine HSCs (Supplementary Figure 1b).…”

“…5 To generate the open reading frame of DEK/CAN, the DEK-and CAN-portions of the DEK/CAN complementary DNA were cloned from peripheral blood mononuclear cells using reverse transcriptase-PCR (RT-PCR) with the following primers containing the indicated restriction sites that are underlined: Dek-Sap-ATG fw 5 0 -GCT CTT CAA TGT CCG CCT CGG-3 0 ; Dek-Mlu1-rev 5 0 -CCA CGC GTC TTT TTG CAA ATC TG-3 0 ; Can-Mlu1 fw 5 0 -GAA GCT CAG ACG CGT GAA ATT CGG CG-3 0 ; Can-Nco-rev 5 0 -CTG AAG AAC CCA TGG CTG TAG A-3 0 ; Can-Nco fw 5 0 -GCC ATG GGT TCT TCA GTG CC-3 0 ; Can-Psh1 rev 5 0 -CTG TGA CAG CAG TCT GCC CAG G-3 0 ; Can-Psh1 fw 5 0 -CAG ACT GCT GTC ACA GCA GC-3 0 ; and Can-EcoR1-TGA rev 5 0 -GCG AAT TCT CAG CTT Figure 1 The t(6;9) fusion protein DEK/CAN and its leukemogenic potential in Sca1 þ /lin À cells (a) Modular organization of the translocation partners in t(6;9) and the resulting DEK/CAN fusion protein and schematic representation of the fusion proteins encoded by der6. The reciprocal CAN/DEK fusion transcript theoretically encoded by der9 is not expressed in t(6;9)-positive patients and, therefore, it is omitted here.…”

“…In fact, the ectopic expression of leukemia-associated fusion proteins, such as PML/RARa, PLZF/RARa or AML-1/ETO in hematopoietic progenitor cells recapitulates the leukemic phenotype. [3][4][5] The t(6;9)(p23;q34) translocation occurs in 1-5% of adult patients with AML. 6 It is associated with a poor prognosis and defines a high-risk group of AML patients.…”

Deacetylase inhibitors modulate proliferation and self-renewal properties of leukemic stem and progenitor cells

“…7,8 AML1/ETO as well as PLZF/RARα have been found to induce the transcription factor c-MYC, 9,10 and ectopic expression of c-MYC in murine bone marrow elicited an aggressive myeloid leukemia by conferring self-renewal capacity to committed myeloid progenitor cells. 11,12 The Polycomb group (PcG) protein BMI1 is essential for the leukemic transformation property of PLZF/RARα and a target gene of c-MYC.…”

“…Ectopic expression of PLZF/RARα or a truncated AML1/ ETO protein (AML1/ETO exon 9) in murine hematopoietic stem cells and transplantation into syngeneic mice recapitulates the leukemic phenotype characterized by a differentiation block by PLZF/RARα or a truncated form of AML1/ETO (AML1/ ETO exon 9), respectively. 7,8 Both leukemia-associated fusion proteins (LAFP) were cloned into proviral constructs and retrovirally transduced into 32D cells. Expression of the fusion proteins was confirmed by western blot (Fig.…”