title: Modulation of p210(BCR-ABL) activity in transduced primary human hematopoietic cells controls lineage programming.
abstract: Retroviral transduction of primary hematopoietic cells with human oncogenes provides a powerful approach to investigating the molecular mechanisms controlling the normal proliferation and differentiation of these cells. Here we show that primitive human CD34(+) cord blood cells, including multipotent as well as granulopoietic- and erythroid-restricted progenitors, can be efficiently transduced with a MSCV-BCR-ABL-IRES-GFP retrovirus, resulting in the sustained expression by their progeny of very high levels of tyrosine phosphorylated p210(BCR-ABL). Interestingly, even in the presence of growth factors that supported the exclusive production of granulopoietic cells from green fluorescent protein (GFP)-transduced control cells, BCR-ABL-transduced progenitor subpopulations generated large numbers of erythropoietin-independent terminally differentiating erythroid cells and reduced numbers of granulopoietic cells. Analyses of individual clones generated by single transduced cells in both semisolid and liquid cultures showed this BCR-ABL-induced erythroid differentiation response to be elicited at a high frequency from all types of transduced CD34(+) cells independent of their apparent prior lineage commitment status. Additional experiments showed that this erythroid differentiation response was largely prevented when the cells were transduced and maintained in the presence of the BCR-ABL-specific tyrosine kinase inhibitor, STI-571. These findings indicate that overexpression of BCR-ABL in primary human hematopoietic cells can activate an erythroid differentiation program in apparently granulopoietic-restricted cells through a BCR-ABL kinase-dependent mechanism, thus providing a new molecular tool for elucidating mechanisms underlying lineage fate determination in human hematopoietic cells and infidelity in human leukemia.
authors.affiliation: Terry Fox Laboratory, British Columbia Cancer Agency, Department of Medicine, University of British Columbia, Vancouver, British Columbia, Canada.
authors.forename: Yves
authors.initials: Y
authors.lastname: Chalandon
authors.forename: Xiaoyan
authors.initials: X
authors.lastname: Jiang
authors.forename: Glen
authors.initials: G
authors.lastname: Hazlewood
authors.forename: Slade
authors.initials: S
authors.lastname: Loutet
authors.forename: Eibhlin
authors.initials: E
authors.lastname: Conneally
authors.forename: Allen
authors.initials: A
authors.lastname: Eaves
authors.forename: Connie
authors.initials: C
authors.lastname: Eaves
mesh_terms: D018952:Antigens, CD34; D001549:Benzamides; D018929:Cell Culture Techniques / Q000379:methods; D002454:Cell Differentiation / Q000187:drug effects; D019070:Cell Lineage / Q000187:drug effects*; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology; D016207:Cytokines / Q000494:pharmacology; D015672:Erythroid Precursor Cells; D016044:Fusion Proteins, bcr-abl / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D006412:Hematopoietic Stem Cells / Q000166:cytology / Q000378:metabolism*; D006801:Humans; D000068877:Imatinib Mesylate; D010879:Piperazines / Q000494:pharmacology; D011743:Pyrimidines / Q000494:pharmacology; D014161:Transduction, Genetic / Q000379:methods
chemical_list: D018952:Antigens, CD34; D001549:Benzamides; D003470:Culture Media; D016207:Cytokines; D010879:Piperazines; D011743:Pyrimidines; D000068877:Imatinib Mesylate; D016044:Fusion Proteins, bcr-abl