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• W2029043405 abstract "Intense activity has been devoted to developing methods for the ex vivo expansion of pluripotent hematopoietic stem cells (HSC) or precursor cells in order to improve clinical transplantation [1.Delaney C, Andrews RG, Bernstein ID. Expansion of hematopoietic stem cells. In: Blume KG, Forman SJ, Appelbaum FR, eds. Thomas' Hematopoietic Cell Transplantation. Blackwell Publishing: In press.Google Scholar]. Such methods for growing cells may prove useful in overcoming the delayed myeloid as well as lymphoid engraftment and the increased early mortality from infectious complications seen with cord blood transplants secondary to inadequate progenitor cell numbers. During the past decade, efforts to expand HSCs have primarily focused on the use of hematopoietic cytokines, and a few-fold increase in HSC number after culture in the presence of particular cytokines and serum-free medium has been demonstrated in murine studies [2.Miller C.L. Eaves C.J. Expansion in vitro of adult murine hematopoietic stem cells with transplantable lympho-myeloid reconstituting ability.Proc Natl Acad Sci U S A. 1997; 94: 13648-13653Crossref PubMed Scopus (210) Google Scholar, 3.Bhatia M. Bonnet D. Kapp U. Wang J.C. Murdoch B. Dick J.E. Quantitative analysis reveals expansion of human hematopoietic repopulating cells after short-term ex vivo culture.J Exp Med. 1997; 186: 619-624Crossref PubMed Scopus (334) Google Scholar]. Using human cord blood cells, Piacibello et al. [4.Piacibello W. Sanavio F. Severino A. et al.Engraftment in nonobese diabetic severe combined immunodeficient mice of human CD34+ cord blood cells after ex vivo expansion: evidence for the amplification and self-renewal of repopulating stem cells.Blood. 1999; 93: 3736-3749Crossref PubMed Google Scholar] obtained expanded numbers of SCID mouse repopulating cells after culture for up to several months under carefully controlled conditions. Ultimately, however, reproducible generation of stem cells and progenitors in numbers sufficient to impact clinical transplantation will require determination of the factors that govern cell-fate decisions of HSC to self-renew or give rise to particular precursor types. Developmental biology studies in systems ranging from D. melanogaster and C. elegans to vertebrates have determined that cell fate is instructed by the integration of several environmental signals that activate a limited number of highly conserved pathways. In addition to the well-studied cytokine-activated receptor tyrosine kinases, such signaling pathways include Notch, wnt (wingless), Hedgehog, and TGF-/BMP. Recent studies have shown that each of these evolutionarily conserved pathways is expressed by hematopoietic precursors, and numerous studies have suggested the potential usefulness of exploiting these pathways in improving stem cell ex vivo culture. The most well studied of these pathways in the context of ex vivo culture for improved stem cell transplantation is the Notch pathway. Notch receptors regulate cell-fate decisions in a wide variety of cell types, often inhibiting particular differentiation programs while permitting either self-renewal or differentiation along an alternate pathway [5.Ohishi K. Varnum-Finney B. Bernstein I.D. The notch pathway: modulation of cell fate decisions in hematopoiesis.Int J Hematol. 2002; 75: 449-566Crossref PubMed Scopus (48) Google Scholar, 6.Artavanis-Tsakonas S. Rand M.D. Lake R.J. Notch signaling: Cell fate control and signal integration in development.Science. 1999; 284: 770-776Crossref PubMed Scopus (4820) Google Scholar]. Initial studies using retrovirus-mediated expression of activated Notch1 demonstrated enhanced self-renewal and immortalized HSC in the context of appropriate cytokines, indicating Notch's potential for enhancing self-renewal in nonmutant precursor cells [7.Varnum-Finney B. Xu L. Brashem-Stein C. et al.Pluripotent, cytokine-dependent, hematopoietic stem cells are immortalized by constitutive Notch1 signaling.Nat Med. 2000; 6: 1278-1281Crossref PubMed Scopus (523) Google Scholar]. Subsequent studies with cell-bound or soluble Notch ligand forms also showed expansion of hematopoietic precursor cell numbers [8.Ohishi K. Varnum-Finney B. Bernstein I.D. Δ-1 enhances marrow and thymus repopulating ability of human CD34+CD38− cord blood cells.J Clin Invest. 2002; 110: 1165-1174Crossref PubMed Scopus (189) Google Scholar, 9.Varnum-Finney B. Brashem-Stein C. Bernstein I.D. Combined effects of Notch signaling and cytokines induce a multiple log increase in precursors with lymphoid and myeloid reconstituting ability.Blood. 2003; 101: 1784-1789Crossref PubMed Scopus (215) Google Scholar, 10.Varnum-Finney B. Purton L.E. Yu M. et al.The notch ligand, jagged-1, influences the development of primitive hematopoietic precursor cells.Blood. 1998; 91: 4084-4091Crossref PubMed Google Scholar, 11.Karanu F. Murdoch B. Gallacher L. et al.The Notch ligand Jagged-1 represents a novel growth factor of human hematopoietic stem cells.J Exp Med. 2000; 192: 1365-1372Crossref PubMed Scopus (361) Google Scholar]. Use of an immobilized Δ1 construct in murine studies led to a multi-log expansion of precursors capable of short-term reconstitution, suggesting the utility of this construct in hastening engraftment [9.Varnum-Finney B. Brashem-Stein C. Bernstein I.D. Combined effects of Notch signaling and cytokines induce a multiple log increase in precursors with lymphoid and myeloid reconstituting ability.Blood. 2003; 101: 1784-1789Crossref PubMed Scopus (215) Google Scholar]. In studies of human cord blood CD34+38− cells, immobilized Δ1 led to a greater than one-log increase in the number of longer-term (>6 months) NOD/SCID repopulating cells, an increase that may be clinically useful for improving both short- and longer-term engraftment ([8.Ohishi K. Varnum-Finney B. Bernstein I.D. Δ-1 enhances marrow and thymus repopulating ability of human CD34+CD38− cord blood cells.J Clin Invest. 2002; 110: 1165-1174Crossref PubMed Scopus (189) Google Scholar], Ohishi et al., unpublished observations). Additional findings with potential clinical significance include the promotion of early T-lymphoid differentiation after culture of hematopoietic precursors with Notch ligand, and the enhanced repopulation of the T cell lineage in addition to the myeloid lineages by HSC cultured with Notch ligand. The ability to generate T-repopulating cells may prove valuable in hastening T-cell engraftment, thereby decreasing the infectious complications of the posttransplant period in patients undergoing HSC transplantation, or in rapidly engrafting SCID patients in the absence of conditioning. Other cell-fate instructive pathways have also been considered as avenues to improving stem cell transplantation. For example, studies of the Wnt pathway showed that overexpression of β-catenin in murine HSC led to enhanced generation of precursors, and purified Wnt 3a together with limiting amounts of SCF expanded the number of repopulating cells detected at 6 weeks [12.Reya T. Duncan A.W. Ailles L. et al.A role for Wnt signalling in self-renewal of haematopoietic stem cells.Nature. 2003; 423: 409-414Crossref PubMed Scopus (1766) Google Scholar, 13.Willert K. Brown J. Danenberg E. et al.Wnt proteins are lipid-modified and can act as stem cell growth factors.Nature. 2003; 423: 448-452Crossref PubMed Scopus (1771) Google Scholar]. Since Wnt was shown to upregulate components of the Notch pathway, the interaction of Notch and Wnt-induced signaling will be of interest. Sonic Hedgehog, another mediator of cell-fate decisions, was found to have modest effects on HSC expansion, while BMP4 conferred a survival effect on HSC [14.Bhatia M. Bonnet D. Wu D. et al.Bone morphogenetic proteins regulate the developmental program of human hematopoietic stem cells.J Exp Med. 1999; 189: 1139-1148Crossref PubMed Scopus (330) Google Scholar, 15.Bhardwaj G. Murdoch B. Wu D. et al.Sonic hedgehog induces the proliferation of primitive human hematopoietic cells via BMP regulation.Nat Immunol. 2001; 2: 172-180Crossref PubMed Scopus (552) Google Scholar]. In summary, multiple signaling pathways known to affect stem cell–fate decisions have been implicated in guiding hematopoietic development, suggesting the potential value of exploiting these pathways in enhancing ex vivo expansion. Moreover, as cell-fate decisions are determined by the integration of multiple signal inputs, it is likely that quantitative approaches will be required to activate multiple pathways simultaneously or sequentially to achieve desired outcomes, possibly including even unlimited HSC ex vivo expansion." @default.
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• W2029043405 title "Hematopoietic stem cell expansion: role of cell-fate instructive pathways" @default.
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