title: Ex vivo expansion of normal and chronic myeloid leukemic stem cells without functional alteration using a NUP98HOXA10homeodomain fusion gene.
abstract: HOX genes have been implicated as regulators of normal and leukemic stem cell functionality, but the extent to which these activities are linked is poorly understood. Previous studies revealed that transduction of primitive mouse hematopoietic cells with a NUP98HOXA10homeodomain (NA10HD) fusion gene enables a subsequent rapid and marked expansion in vitro of hematopoietic stem cell numbers without causing their transformation or deregulated expansion in vivo. To determine whether forced expression of NA10HD in primitive human cells would have a similar effect, we compared the number of long-term culture-initiating cells (LTC-ICs) present in cultures of lenti-NA10HD versus control virus-transduced CD34(+) cells originally isolated from human cord blood and chronic phase (CP) chronic myeloid leukemia (CML) patients. We found that NA10HD greatly increases outputs of both normal and Ph(+)/BCR-ABL(+) LTC-ICs, and this effect is particularly pronounced in cultures containing growth factor-producing feeders. Interestingly, NA10HD did not affect the initial cell cycle kinetics of the transduced cells nor their subsequent differentiation. Moreover, immunodeficient mice repopulated with NA10HD-transduced CP-CML cells for more than 8 months showed no evidence of altered behavior. Thus, NA10HD provides a novel tool to enhance both normal and CP-CML stem cell expansion in vitro, without apparently altering other properties.
authors.affiliation: Terry Fox Laboratory, British Columbia Cancer Agency, Vancouver, British Columbia, Canada.
authors.forename: I
authors.initials: I
authors.lastname: Sloma
authors.forename: S
authors.initials: S
authors.lastname: Imren
authors.forename: P A
authors.initials: PA
authors.lastname: Beer
authors.forename: Y
authors.initials: Y
authors.lastname: Zhao
authors.forename: V
authors.initials: V
authors.lastname: Lecault
authors.forename: D
authors.initials: D
authors.lastname: Leung
authors.forename: K
authors.initials: K
authors.lastname: Raghuram
authors.forename: C
authors.initials: C
authors.lastname: Brimacombe
authors.forename: K
authors.initials: K
authors.lastname: Lambie
authors.forename: J
authors.initials: J
authors.lastname: Piret
authors.forename: C
authors.initials: C
authors.lastname: Hansen
authors.forename: R K
authors.initials: RK
authors.lastname: Humphries
authors.forename: C J
authors.initials: CJ
authors.lastname: Eaves
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D015153:Blotting, Western; D002453:Cell Cycle; D002454:Cell Differentiation; D049109:Cell Proliferation; D002478:Cells, Cultured; D003114:Colony-Forming Units Assay; D005312:Fetal Blood / Q000166:cytology / Q000378:metabolism; D016044:Fusion Proteins, bcr-abl / Q000235:genetics; D006412:Hematopoietic Stem Cells / Q000166:cytology* / Q000378:metabolism; D000080905:Homeobox A10 Proteins; D018398:Homeodomain Proteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D015466:Leukemia, Myeloid, Chronic-Phase / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D051379:Mice; D016688:Mice, Inbred NOD; D016513:Mice, SCID; D014411:Neoplastic Stem Cells / Q000166:cytology* / Q000378:metabolism; D028861:Nuclear Pore Complex Proteins / Q000235:genetics* / Q000378:metabolism; D015514:Oncogene Proteins, Fusion; D012333:RNA, Messenger / Q000235:genetics; D060888:Real-Time Polymerase Chain Reaction; D015375:Receptors, Interleukin-2; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014183:Transplantation, Heterologous
chemical_list: D018952:Antigens, CD34; D000080905:Homeobox A10 Proteins; D018398:Homeodomain Proteins; D028861:Nuclear Pore Complex Proteins; C465451:Nup98 protein, human; D015514:Oncogene Proteins, Fusion; D012333:RNA, Messenger; D015375:Receptors, Interleukin-2; C069634:HOXA10 protein, human; D016044:Fusion Proteins, bcr-abl