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HEMATOPOIESIS
From The Terry Fox Laboratory, British Columbia Cancer
Agency, and the Departments of Medicine and Medical Genetics,
University of British Columbia, Vancouver, BC, Canada.
Several studies point to multiple members of the Hox transcription
factor family as playing key roles in normal hematopoietic development,
and they link the imbalanced expression of these transcription factors,
in particular of the Abd-like A cluster HOX
genes HOXA9 and HOXA10, to
leukemogenesis. To test directly the hypothesis that HOXA10
is involved in human hematopoietic development, the gene was
retrovirally overexpressed in human highly purified
CD34+/GFP+ hematopoietic progenitor
cells derived from cord blood or fetal liver sources, and the impact of
aberrant gene expression was analyzed on differentiation and
proliferation in vitro and in vivo. HOXA10 misexpression
profoundly impaired myeloid differentiation with a higher yield of
blast cells in liquid culture and a greater than 100-fold increased
generation of blast colonies after in vitro expansion or after
replating of primary colonies first plated in methylcellulose directly
after transduction (P < .01). Furthermore, aberrant
HOXA10 expression almost completely blocked erythroid differentiation in methylcellulose (P < .02).
HOXA10 deregulation also severely perturbed the
differentiation of human progenitors in vivo, reducing B-cell
development by 70% in repopulated NOD/SCID mice and enhancing
myelopoiesis in the transduced compartment. The data provide evidence
that the balanced expression of HOXA10 is pivotal for
normal human hematopoietic development and that aberrant expression of
the gene contributes to impaired differentiation and increased
proliferation of human hematopoietic progenitor cells. These results
also provide a framework to initiate more detailed analyses of
HOX regulatory domains and HOX cofactors in the
human system in vitro and in vivo.
(Blood. 2001;97:2286-2292) HOX homeobox genes were first
recognized as an evolutionarily conserved family of transcription
factors critical to the control of early embryonic development. More
recently, many of these genes have been implicated in the regulation of
both normal and leukemic hematopoiesis.1,2 Examples of the
latter include the frequent involvement of PBX1, a cofactor of
HOX function, or MLL, a putative upstream regulator of
HOX gene expression and of HOXA9 in human leukemia-associated fusion genes.3-7 More recently, we and
others have shown that the leukemic blasts from many patients with
acute myeloid leukemia (AML) show an aberrant pattern of
HOXA10 expression, further supporting the concept that
dysregulated HOX gene expression may be a general feature of
this malignancy.8,9 Notably, the expression of
HOXA9 has been identified as one of the most consistent
diagnostic markers of AML in humans and as the only single gene
expression marker of more than 6800 cDNAs tested by DNA micro-array
analysis that correlated with clinical outcome.10 Additional evidence of the leukemogenic potential of deregulated HOX gene expression and, in particular, of certain members
of the A cluster have come from analysis of mouse models of leukemia. In the BXH2 mouse, the development of AML is linked to the activation of HOXA9 or HOXA7 by retroviral insertion, often
in concert with activation of the Meis cofactor.11
Retrovirus-mediated overexpression of HOXA10 or
HOXA9 in murine hematopoietic cells also leads to AML in
mice transplanted with these cells after an initial "latency" period, suggesting the additional involvement of certain
co-operating oncogenes.12,13 The relevance of normal
HOX gene expression for hematopoietic development is
demonstrated by the HOXA9 Both HOXA9 and HOXA10 belong to the group of
so-called Abdominal B-like genes, which are homologous to
the most 5' gene Abdominal B (AbdB), initially
identified and characterized in the Drosophila homeotic
complex. The Abd-like genes share a number of distinguishing
features, such as a conserved tryptophan motif mediating the binding of
the Pbx protein, differences at various positions in the flexible
N-terminal arm of the DNA-binding homeodomain, preferential recognition
of a TTAT core in contrast to the Antennapedia (Antp) group
of homeodomains, and an insensitivity to activation by retinoic
acid.15-18 HOXA10 and HOXA9 are
expressed in the most primitive CD34+ cell compartment of
normal adult human bone marrow cells, and their expression is virtually
extinguished in the more prevalent differentiating CD34 To test this hypothesis directly in primitive human hematopoietic
cells, we initiated the present study. We focused on the analysis of
the effects of HOXA10 overexpression in the progeny of human
cord blood or fetal liver cells transduced in vitro with a retroviral
vector encoding the human HOXA10 cDNA linked by an internal
ribosomal entry site (IRES) element to enhanced green fluorescent
protein. Severe perturbations of the proliferation and differentiation
of the transduced cells, detectable both in vitro and in vivo in
repopulated NOD/SCID mice, were seen. These findings establish the
feasibility of using this approach to dissect the consequences of
altered expression of specific HOX genes on primitive human
hematopoietic cell behavior and underline the importance of normal
expression of the Abd-like HOXA10 gene for proper
human hematopoietic development.
Retroviral constructs
Human cells
Retroviral transduction of human hematopoietic progenitor cells Lin cord blood or fetal liver cells were
transduced as previously described.21 Briefly, cells at
2 × 105/mL were prestimulated for 48 hours in Iscove
minimum Dulbecco medium (IMDM) containing a serum substitute (BIT;
Stemcell Technologies), 104 M mercaptoethanol (Sigma
Chemical, St Louis, MO), and 40 µg/mL low-density lipoproteins
(Sigma) supplemented with the following recombinant human cytokines:
100 ng/mL Flt-3 ligand (Immunex, Seattle, WA) 100 ng/mL steel factor
(SF, prepared and purified in the Terry Fox Laboratory) 20 ng/mL
interleukin-3 (IL-3) (Novartis, Basel, Switzerland), 20 ng/mL IL-6
(Cangene, Mississauga, ON), and 20 ng/mL granulocyte-colony stimulating
factor (G-CSF; Stem Cell). After 48 hours, cells were harvested and
resuspended in filtered virus-containing medium supplemented with the
same cytokine combination and protamine sulfate (5 µg/mL) on Petri
dishes that had been precoated with 5 µg/cm2 fibronectin
(Sigma) and preloaded twice with virus-containing medium, each time for
30 minutes, as described.21 This procedure was repeated on
the next 2 consecutive days, for a total of 3 infections. For
subsequent in vitro studies, cells were transferred to fresh serum-free
medium plus cytokines and incubated for an additional 48 hours before
they were stained with Cy5-labeled anti-CD34 antibody (Becton
Dickinson, San Jose, CA). Transduced GFP+/CD34+
cells were then quantitated and isolated using a 3-laser Facstar Plus
(Becton Dickinson). Immunodeficient mice repopulated with transduced
cells were injected without preselection less than 24 hours after the
third day of exposure to virus.
In vitro progenitor assays Assays for in vitro colony-forming cells (CFCs) were carried out in methylcellulose cultures (GF H4434; Stemcell Technologies) supplemented with 50 ng/mL human SF, 20 ng/mL each of human IL-3, IL-6, GM-CSF (Novartis), and G-CSF, and 3 U/mL erythropoietin (Stemcell Technologies), as described previously.24 Secondary progenitor assays were performed by replating aliquots of cells obtained by harvesting all the cells in 14-day-old primary assays. Cell morphology was assessed by Wright-Giemsa-stained smears of individually plucked colonies. Long-term culture-initiating cell assays (LTC-ICs) were carried out as previously described using pre-established irradiated murine fibroblasts genetically engineered to produce human IL-3, G-CSF, and SF.24 For in vitro liquid expansion assays, cells were cultured in the same serum-free medium described above. The types of cells present in these cultures at various time points was determined by staining cytospin preparations with Wright-Giemsa and by plating cells in CFC assays.Mice NOD/LtSz-scid/scid (NOD/SCID) mice were bred and maintained in micro-isolators under sterile conditions in the animal facility of the BC Cancer Research Centre, as previously described.25 Eight- to 10-week-old mice were irradiated with 350 cGy from a cesium Cs 137 source less than 24 hours before human cells were injected. Engraftment and lineage differentiation were analyzed 8 weeks after transplantation by the aspiration of cells from the femurs under light anesthesia. Mice were humanely killed 12 weeks after transplantation, and the bone marrow was obtained from femurs and tibias. Cell suspensions were prepared in cold Hanks balanced salt solution, supplemented with 5% pooled normal human serum (Stemcell Technologies) for fluorescence-activated cell sorter (FACS) and progenitor analyses. The absolute number of human engrafted cells per mouse was calculated based on the recovered cells obtained from femurs and tibias and represented 25% of the total marrow.Human lymphomyeloid engraftment and lineage representation of cells
obtained from the mice 8 and 12 weeks after transplantation were
determined as previously described.21,26 Briefly, red cells were lysed with 7% ammonium chloride (Stemcell Technologies), washed, labeled with 1% propidium iodine (PI; Sigma), and incubated on
ice for 10 minutes with human serum supplemented with an anti-mouse IgG
Fc receptor antibody (2.4G2; Systemix, Palo Alto, CA) to block mouse Fc
receptors and to minimize nonspecific binding. Separate aliquots were
then incubated for 30 minutes on ice with a mouse isotype-matched
control antibody (Becton Dickinson) or the following antibodies against
human antigens: antihuman CD34-Cy5,27
CD19-phycoerythrin (PE; Becton Dickinson), CD15-PE (Pharmingen,
Ontario, Canada), and CD45-PE (Pharmingen). Viable
(PI Statistical analysis Data were statistically tested using the t test for dependent or independent samples (Statistica 5.1; Statsoft, Tulsa, OK). Differences with P < .05 were considered statistically significant.
Efficient retroviral transduction of HOXA10 in human hematopoietic progenitor cells To facilitate the selection and tracking of retrovirally transduced cells by FACS, a bicistronic vector for the long terminal repeat (LTR)-driven expression of HOXA10 and GFP was constructed in the MSCV viral backbone (A10-GFP virus, Figure 1A). The analogous vector containing GFP alone (GFP virus) was used as a control. Full-length proviral integration and transcription were confirmed by Southern (Figure 1B) and Northern blot (Figure 1C) analyses, respectively, of stable PG13 producer cells, transduced hematopoietic target cell lines, K562, MO7E, and transduced primary cord blood cells. Fifty-four percent ( ± 17%) and 70% ( ± 12%) of low-density Lin
cord blood cells exposed to the A10-GFP vector or the GFP vector, respectively, were found to be GFP+ when examined by FACS,
demonstrating the high efficiency of gene transfer by both
vectors.
HOXA10-transduced cord blood cells produce increased numbers of primitive cells in vitro An immediate effect of HOXA10 overexpression on transduced cord blood cells was revealed by comparing the numbers and morphology of cells generated from CD34+ GFP+ cells isolated by FACS 48 hours after their transduction with the A10-GFP or GFP vectors and placed in serum-free liquid cultures containing fetal liver, SF, IL-3, IL-6, and G-CSF. One week later, the total number of cells present in cultures initiated with HOXA10-transduced cord blood cells was approximately 1.7-fold higher than in parallel cultures initiated with GFP-transduced cord blood cells. Mean expansion (range) over input was 49 (17 to 109)-fold versus 33 (6.5 to 77)-fold, respectively; n = 4). This increased expansion was associated with an even greater (2.5-fold) increased production of blast cells. The mean proportion was 52% ± 18% vs 30% ± 11%, resulting in absolute numbers of 6.8 × 105 (0.5 × 105 to 18.6 × 105) versus 2.7 × 105 (0.06 × 105 to 7.1 × 105) blasts, respectively, per 1 × 104 CD34+GFP+ cells initially placed in culture. Analysis of the hematopoietic progenitor content of serum-free suspension cultures after 2 weeks showed that the mean absolute number of CFCs was 21-fold higher. The number of blast colonies was 125-fold higher in the cultures initiated with HOXA10-transduced cells than in the control cultures (n = 2; HOXA10, 29 and 1600 blast colonies, respectively; GFP, 0 and 13 blast colonies, respectively) (Figure 2). These results suggested that HOXA10 overexpression delayed myeloid differentiation.
HOXA10 induces alterations in cord blood progenitor cell differentiation in vitro To determine whether dysregulated HOXA10 expression would also affect the proliferation and differentiation activity of progenitors detectable in other assay systems, CD34+/GFP+ cord blood cells isolated by FACS 48 hours after transduction were either plated directly into methylcellulose cultures or first cultured for 6 weeks on murine stromal fibroblast feeder layers engineered to produce human SF, IL-3, and G-CSF. Colony numbers obtained from direct CFC assays of cells from 6 independent transduction experiments are summarized in Figure 3A. The total number of CFCs detected immediately after transduction was reduced for the HOXA10-transduced cells compared to the control GFP-transduced cells. This was because of an almost complete block of erythroid differentiation in the colonies generated by HOXA10-transduced cells (more than 85% reduction in the number of "pure" erythroid colonies; n = 6; P = .017; 54% reduction of mixed erythroid-"myeloid" colonies by comparison to the control GFP-transduced cells). Replating studies showed that, as expected, only small numbers of colonies were obtained when control primary CFC cultures were assayed in secondary cultures (median, 30 secondary colonies per 400 primary CD34+ GFP+ cells plated into the primary CFC assays) (Figure 3B). In contrast, more than 100-fold higher numbers of secondary colonies were generated on the replating of cells from primary CFC assays of HOXA10-transduced cells (median, 3700 colonies per 400 initially plated CD34+GFP+; P < .01). Moreover, Wright-Giemsa staining showed approximately 80% of these HOXA10-derived secondary colonies to consist morphologically of blasts (Figure 3C). In 2 of 4 experiments, HOXA10-transduced cells obtained from the secondary colonies were able to form tertiary colonies, though at somewhat reduced numbers, and all tertiary colonies consisted of blast cells as assessed by morphology.
When the initially transduced CD34+GFP+ cells were cultured under LTC-IC assay conditions, the number of CFCs present 6 weeks later was similar for both HOXA10 and control GFP+ input populations. However, most (68% ± 8%) of colonies generated by HOXA10-transduced cells maintained under these conditions again consisted of blasts or granulocyte precursors, whereas the CFCs obtained from LTC-IC assays of control cells produced mainly mature granulocyte-macrophage colonies (99.5% ± 0.25%). These studies indicate the ability of HOXA10 overexpression to significantly impair myeloid differentiation, particularly the terminal aspects of erythroid differentiation. HOXA10 perturbs the output of lymphoid cells by in vivo repopulating human progenitor cells To determine whether overexpression of HOXA10 would affect the proliferation and differentiation of cord blood cells able to engraft the marrow of NOD/SCID mice, cord blood or fetal liver cells were injected into mice immediately after infection, without any preselection of CD34+ GFP+ cells. Each mouse was injected with the progeny of an original input of Lin
cells containing 105 CD34+ cells. Engraftment
with human cells was first assessed by FACS analysis of bone marrow
cells obtained by femoral bone marrow aspirate 8 weeks after
transplantation and again 12 weeks after, when the mice were killed. At
8 weeks after transplantation, both human lymphoid
(CD34CD19+) and human myeloid
(CD15+) cells could be detected in 13 of 14 animals
injected with A10-GFP cells and in 6 of 14 mice injected with control
GFP cells. Moreover, in all cases, human lymphomyeloid engraftment was
evident among both the GFP+ and the GFP
cells. Four weeks later, 11 of 14 animals in the HOXA10
group and 7 of 14 in the GFP control group showed lymphomyeloid
engraftment, with a median frequency in human cells of 18.5%
(0.7%-66%) and 23% (5%-66%), respectively. Twelve percent
(2%-48%) of the engrafted human cells were transduced in the mice
transplanted with A10-GFP cells versus 12% (2%-89%) in the mice
transplanted with control GFP cells.
Although overexpression of HOXA10 in human NOD/SCID
repopulating cells was compatible with lymphomyeloid engraftment, the proportion and absolute yield of human lymphoid and myeloid cells produced in the recipients of HOXA10-transduced cells were
significantly altered. In both the GFP+ and the
GFP
Several lines of evidence have shown that the homeobox family plays a critical role in normal hematopoietic development and that their imbalanced expression is linked to leukemogenesis. In particular, the 5'-located A cluster HOX genes, such as HOXA9 and HOXA10, are associated with the development of leukemia by the molecular characterization of leukemia-specific fusion genes or by studies in murine transplantation models, using bone marrow cells engineered to overexpress the HOXA10 or HOXA9 gene.3,4,12,13 The precise regulatory role of these Abd-like A cluster HOX genes in human hematopoietic development has not yet been dissected. To test the hypothesis that HOXA10 is critically involved in early hematopoietic differentiation in humans, we aberrantly expressed this gene in human hematopoietic progenitor cells and studied the effect of misexpression on hematopoietic development in vitro and in vivo. For this, we exploited recent advances in the technique of retroviral gene transfer that allow the transduction of human hematopoietic progenitor cells, including those that repopulate NOD/SCID mice with high efficiency.21 Thus, it was possible to analyze the effect of deregulated HOX gene expression in vivo for the first time in the human system. In vitro studies revealed striking effects of HOXA10
overexpresssion on cell expansion in liquid culture or cell
proliferation of clonogenic progenitors in methylcellulose. In
parallel, HOXA10 profoundly altered the normal
differentiation program of early progenitors and impaired myeloid
differentiation: it increased the yield of blast cells in liquid
culture, induced a greater than 100-fold enhanced generation of blast
colonies after in vitro expansion or after replating of methylcellulose
colony assays, and led to an almost complete block of erythroid colony
formation. Furthermore, it perturbed the differentiation of LTC-IC such
that most of their clonogenic progeny detected after 6 weeks in culture were CFU-blast. These observations probably highlight key regulatory features of this gene In our study we also analyzed the impact of HOXA10 deregulation on the differentiation of human progenitors in vivo in sublethally irradiated NOD/SCID mice. These experiments demonstrated a substantial impairment in B-cell development in mice transplanted with cells overexpressing the gene. So far, there are no data on the effect of other HOX genes on B-cell differentiation in the human system. However, data from murine transplantation models demonstrated that both HOXA10 and HOXA9 are detrimental to B-cell development in vivo.12,13 There are no precise data concerning at which stage of B-cell development the block occurs, but pre-B colony formation assays suggest that the development of murine IL-7-sensitive pre-B cells is impaired by HOXA10 overexpression.12 The impact of HOXA10 on early B-cell progenitors was confirmed by recent results suggesting that HOXA10 overexpression critically impairs the transition from the pro-B to the pre-B-cell stage, as defined by Hardy in HOXA10 transgenic mice.33,34 In addition, the in vivo studies confirmed that HOXA10 overexpression perturbed normal myeloid development: misexpression of the gene had a myeloproliferative effect with an increased frequency of CFCs in the human CD34+ compartment, an impairment of terminal myeloid differentiation ex vivo, and a competitive growth advantage of transduced myeloid cells in a subgroup of mice. However, we did not observe frank signs of a myeloproliferative syndrome or of leukemia in the animals. It may be that other HOX cofactors are required for accelerated leukemogenesis. This was shown for HOXA9 by a shortened latency period to the development of leukemia in mice transplanted with cells overexpressing both HOXA9 and its cofactor, MEIS1, as compared with the control animals transplanted with cells overexpressing HOXA9 alone. In addition, structure-function analysis has demonstrated the importance of the MEIS- and PBX-interacting domains for HOXA9-mediated immortalization.13,35 However, it is likely that the limited proliferative potential demonstrated by most human hematopoietic cells in the NOD/SCID mouse model have made it difficult to detect a HOXA10-induced myeloproliferative process in these animals. With regard to the impact of HOXA10 on erythroid differentiation in vivo, we noticed a substantial decrease in the median proportion of erythroid glycophorin A+ cells in the HOXA10-transduced human cell population compared with the GFP+ cell population in the control mice (HOXA10, 0.5%; GFP, 12.9%; data not shown). However, the frequency of glycophorin A+ cells was low, and xenotransplant models, which allow a more efficient erythroid differentiation of human engrafted cells, will help to analyze more precisely the effect of HOXA10 on erythropoiesis in vivo. In summary, these data demonstrate that regulated HOXA10 expression may be pivotal for normal progenitor development and that aberrant expression of this gene results in increased proliferation and impaired differentiation of normal human hematopoietic cells. When combined with additional genetic events, these changes may be key factors in HOXA cluster-linked leukemogenesis. This in vitro and in vivo study now allows the study of these aspects in detail. It opens the door to assess more precisely the role of HOX cofactors, such as PBX1 or MEIS1, in HOXA10-induced perturbations of hematopoiesis and to perform structure-function analysis of putative HOXA10 regulatory domains in the experimental human systems described here.
Submitted October 6, 2000; accepted December 19, 2000.
Supported by the National Cancer Institute of Canada, with funds from the Canadian Cancer Society and the Terry Fox Run, and by the National Institutes of Health (grant DK48642). C.B. is supported by a grant from the Deutsche Forschungsgesellschaft (Bonn, Germany). M.F.-B. is supported by a grant from the Deutsche Krebshilfe (Bonn, Germany).
The publication costs of this article were defrayed in part by page charge payment. Therefore, and solely to indicate this fact, this article is hereby marked "advertisement" in accordance with 18 U.S.C. section 1734.
Reprints: R. Keith Humphries, The Terry Fox Laboratory, British Columbia Cancer Agency, 10th Ave West, Vancouver, BC, Canada V5ZIL3; e-mail: keith{at}terryfox.ubc.ca.
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 V. P. S. Rawat, S. Thoene, V. M. Naidu, N. Arseni, B. Heilmeier, K. Metzeler, K. Petropoulos, A. Deshpande, L. Quintanilla-Martinez, S. K. Bohlander, et al. Overexpression of CDX2 perturbs HOX gene expression in murine progenitors depending on its N-terminal domain and is closely correlated with deregulated HOX gene expression in human acute myeloid leukemia Blood, January 1, 2008; 111(1): 309 - 319. [Abstract] [Full Text] [PDF]
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 D. Caudell, Z. Zhang, Y. J. Chung, and P. D. Aplan Expression of a CALM-AF10 Fusion Gene Leads to Hoxa Cluster Overexpression and Acute Leukemia in Transgenic Mice Cancer Res., September 1, 2007; 67(17): 8022 - 8031. [Abstract] [Full Text] [PDF]
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 H. Wang, Y. Lu, W. Huang, E. T. Papoutsakis, P. Fuhrken, and E. A. Eklund HoxA10 Activates Transcription of the Gene Encoding Mitogen-activated Protein Kinase Phosphatase 2 (Mkp2) in Myeloid Cells J. Biol. Chem., June 1, 2007; 282(22): 16164 - 16176. [Abstract] [Full Text] [PDF]
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M. Magnusson, A. C. M. Brun, N. Miyake, J. Larsson, M. Ehinger, J. M. Bjornsson, A. Wutz, M. Sigvardsson, and S. Karlsson HOXA10 is a critical regulator for hematopoietic stem cells and erythroid/megakaryocyte development Blood, May 1, 2007; 109(9): 3687 - 3696. [Abstract] [Full Text] [PDF]
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 J. A. Kennedy, F. Barabe, B. J. Patterson, J. Bayani, J. A. Squire, D. L. Barber, and J. E. Dick Expression of TEL-JAK2 in primary human hematopoietic cells drives erythropoietin-independent erythropoiesis and induces myelofibrosis in vivo PNAS, November 7, 2006; 103(45): 16930 - 16935. [Abstract] [Full Text] [PDF]
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 L. Mar and P. A. Hoodless Embryonic fibroblasts from mice lacking tgif were defective in cell cycling. Mol. Cell. Biol., June 1, 2006; 26(11): 4302 - 4310. [Abstract] [Full Text] [PDF]
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H. B. Sieburg, R. H. Cho, B. Dykstra, N. Uchida, C. J. Eaves, and C. E. Muller-Sieburg The hematopoietic stem compartment consists of a limited number of discrete stem cell subsets Blood, March 15, 2006; 107(6): 2311 - 2316. [Abstract] [Full Text] [PDF]
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 H. Du, G. S. Daftary, S. I. Lalwani, and H. S. Taylor Direct Regulation of HOXA10 by 1,25-(OH)2D3 in Human Myelomonocytic Cells and Human Endometrial Stromal Cells Mol. Endocrinol., September 1, 2005; 19(9): 2222 - 2233. [Abstract] [Full Text] [PDF]
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K. Chadwick, F. Shojaei, L. Gallacher, and M. Bhatia Smad7 alters cell fate decisions of human hematopoietic repopulating cells Blood, March 1, 2005; 105(5): 1905 - 1915. [Abstract] [Full Text] [PDF]
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A. C. M. Brun, J. M. Bjornsson, M. Magnusson, N. Larsson, P. Leveen, M. Ehinger, E. Nilsson, and S. Karlsson Hoxb4-deficient mice undergo normal hematopoietic development but exhibit a mild proliferation defect in hematopoietic stem cells Blood, June 1, 2004; 103(11): 4126 - 4133. [Abstract] [Full Text] [PDF]
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N. Pineault, C. Abramovich, H. Ohta, and R. K. Humphries Differential and Common Leukemogenic Potentials of Multiple NUP98-Hox Fusion Proteins Alone or with Meis1 Mol. Cell. Biol., March 1, 2004; 24(5): 1907 - 1917. [Abstract] [Full Text] [PDF]
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V. P. S. Rawat, M. Cusan, A. Deshpande, W. Hiddemann, L. Quintanilla-Martinez, R. K. Humphries, S. K. Bohlander, M. Feuring-Buske, and C. Buske Ectopic expression of the homeobox gene Cdx2 is the transforming event in a mouse model of t(12;13)(p13;q12) acute myeloid leukemia PNAS, January 20, 2004; 101(3): 817 - 822. [Abstract] [Full Text] [PDF]
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J. M. Bjornsson, N. Larsson, A. C. M. Brun, M. Magnusson, E. Andersson, P. Lundstrom, J. Larsson, E. Repetowska, M. Ehinger, R. K. Humphries, et al. Reduced Proliferative Capacity of Hematopoietic Stem Cells Deficient in Hoxb3 and Hoxb4 Mol. Cell. Biol., June 1, 2003; 23(11): 3872 - 3883. [Abstract] [Full Text] [PDF]
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N. Pineault, C. Buske, M. Feuring-Buske, C. Abramovich, P. Rosten, D. E. Hogge, P. D. Aplan, and R. K. Humphries Induction of acute myeloid leukemia in mice by the human leukemia-specific fusion gene NUP98-HOXD13 in concert with Meis1 Blood, June 1, 2003; 101(11): 4529 - 4538. [Abstract] [Full Text] [PDF]
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B. Schiedlmeier, H. Klump, E. Will, G. Arman-Kalcek, Z. Li, Z. Wang, A. Rimek, J. Friel, C. Baum, and W. Ostertag High-level ectopic HOXB4 expression confers a profound in vivo competitive growth advantage on human cord blood CD34+ cells, but impairs lymphomyeloid differentiation Blood, March 1, 2003; 101(5): 1759 - 1768. [Abstract] [Full Text] [PDF]
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A. Thompson, M. F. Quinn, D. Grimwade, C. M. O'Neill, M. R. Ahmed, S. Grimes, M. F. McMullin, F. Cotter, and T. R. J. Lappin Global down-regulation of HOX gene expression in PML-RARalpha + acute promyelocytic leukemia identified by small-array real-time PCR Blood, February 15, 2003; 101(4): 1558 - 1565. [Abstract] [Full Text] [PDF]
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 N. B. Ivanova, J. T. Dimos, C. Schaniel, J. A. Hackney, K. A. Moore, and I. R. Lemischka A Stem Cell Molecular Signature Science, October 18, 2002; 298(5593): 601 - 604. [Abstract] [Full Text] [PDF]
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C. Buske, M. Feuring-Buske, C. Abramovich, K. Spiekermann, C. J. Eaves, L. Coulombel, G. Sauvageau, D. E. Hogge, and R. K. Humphries Deregulated expression of HOXB4 enhances the primitive growth activity of human hematopoietic cells Blood, July 18, 2002; 100(3): 862 - 868. [Abstract] [Full Text] [PDF]
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T. Taghon, F. Stolz, M. De Smedt, M. Cnockaert, B. Verhasselt, J. Plum, and G. Leclercq HOX-A10 regulates hematopoietic lineage commitment: evidence for a monocyte-specific transcription factor Blood, February 15, 2002; 99(4): 1197 - 1204. [Abstract] [Full Text] [PDF]
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Top
Abstract
Introduction
-actin gene.
3.7 × 106 CD19+ cells (±2.7)
versus 11.9 × 106 cells (±9.1) per mouse, respectively,
with a proportional predominance of human myeloid cells in this
compartment (P < .02) (Table
