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Differentiation

Lentiviral PU.1 overexpression restores differentiation in myeloid leukemic blasts

Leukemia volume 21, pages 1050–1059 (2007)Cite this article

Abstract

PU.1, a transcription factor of the ETS family, plays a pivotal role in normal hematopoiesis, and particularly in myeloid differentiation. Altered PU.1 function is possibly implicated in leukemogenesis, as PU.1 gene mutations were identified in some patients with acute myeloid leukemia (AML) and as several oncogenic products (AML1-ETO, promyelocytic leukemia-retinoic acid receptor alpha, FMS-like receptor tyrosine kinase 3 internal tandem duplication) are associated with PU.1 downregulation. To demonstrate directly a role of PU.1 in the blocked differentiation of leukemic blasts, we transduced cells from myeloid cell lines and primary blasts from AML patients with a lentivector encoding PU.1. In NB4 cells we obtained increases in PU.1 mRNA and protein, comparable to increases obtained with all-trans retinoic acid-stimulation. Transduced cells showed increased myelomonocytic surface antigen expression, decreased proliferation rates and increased apoptosis. Similar results were obtained in primary AML blasts from 12 patients. These phenotypic changes are characteristic of restored blast differentiation. PU.1 should therefore constitute an interesting target for therapeutic intervention in AML.

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Abbreviations

AML:

acute myeloid leukemia

LTR:

long terminal repeat

EF1:

elongation factor 1

cPPT:

central polypurine tract

IRES:

internal ribosome entry site

GFP:

green fluorescent protein

WPRE:

post-transcriptional regulatory element of woodchuck hepatitis virus

SIN:

self-inactivating modification

CMV:

cytomegalovirus

PML-RARA:

promyelocytic leukemia-retinoic acid receptor alpha

FLT3-ITD:

FMS-like receptor tyrosine kinase 3 internal tandem duplication

CAT:

Chloramphenicol acetyl transferase

ATRA:

all-trans retinoic acid

HSC:

hematopoietic stem cells

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Acknowledgements

We thank E Contassot for providing myeloid cell lines, M Barnet for lentivirus productions, D Trono and M Wiznerowicz for providing lentivectors, F Moreau-Gachelin for her gift of PU.1cDNA and PU.1RE, D Wohlwend and S Bissat for cell sorting. This work was supported by Swiss Cancer League Grant No. OCS-01343-2003 and OCS-01781-08-2005.

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Authors and Affiliations

  1. Division of Hematology, University Hospital Geneva, Geneva, Switzerland

    S Durual, A Rideau, S Ruault-Jungblut, P Beris & T Matthes

  2. Division of Clinical Pathology, University Hospital Geneva, Geneva, Switzerland

    D Cossali

  3. Department of Dermatology and Venerology, University Hospital Geneva, Geneva, Switzerland

    V Piguet

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  1. S Durual
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Correspondence to T Matthes.

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Supplementary Information accompanies the paper on the Leukemia website (http://www.nature.com/leu)

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Durual, S., Rideau, A., Ruault-Jungblut, S. et al. Lentiviral PU.1 overexpression restores differentiation in myeloid leukemic blasts. Leukemia 21, 1050–1059 (2007). https://doi.org/10.1038/sj.leu.2404645

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