title: Competitive engraftment of hematopoietic stem cells genetically modified with a truncated erythropoietin receptor.
abstract: Transplantation of genetically modified hematopoietic stem cells (HSCs) has therapeutic potential for a variety of blood genetic disorders. Engraftment of HSCs, however, requires toxic myeloablative treatments, which render this approach questionable for non-life-threatening disorders. A potential alternative is the use of transgenes, which allows positive selection of HSCs in vivo. We used retroviral vectors to express a truncated derivative of the erythropoietin receptor (tEpoR) in murine and human hematopoietic cells. Murine HSCs expressing tEpoR at different levels (1500 to 13,000 receptors/cell) acquire a competitive repopulation capacity in vivo upon transplantation into fully or partially myeloablated co-isogenic mouse recipients. Long-term analysis of transplanted mice showed that expression of tEpoR at paraphysiological levels (approximately 1500 receptors/cell) has no effect on steady-state hematopoiesis and induces no further expansion of transduced cells after the engraftment period. Human cord blood-derived CD34+ stem/progenitor cells transduced with a lentiviral vector expressing tEpoR expand their clonogenic capacity in vitro, and significantly increase their marrow repopulation capacity upon xenotransplantation into sublethally irradiated NOD-SCID mice, with no alteration in their phenotype, survival, and differentiation properties. These data indicate that expression of tEpoR is an effective strategy to promote selective engraftment of genetically modified HSCs upon transplantation in both myeloablative and nonmyeloablative conditions, without the use of toxic drugs for selection.
authors.affiliation: Department of Biomedical Sciences, University of Modena and Reggio Emilia, 41100 Modena, Italy.
authors.forename: Fabrizia
authors.initials: F
authors.lastname: Urbinati
authors.forename: Francesco
authors.initials: F
authors.lastname: Lotti
authors.forename: Giulia
authors.initials: G
authors.lastname: Facchini
authors.forename: Monica
authors.initials: M
authors.lastname: Montanari
authors.forename: Giuliana
authors.initials: G
authors.lastname: Ferrari
authors.forename: Fulvio
authors.initials: F
authors.lastname: Mavilio
authors.forename: Alexis
authors.initials: A
authors.lastname: Grande
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D002478:Cells, Cultured; D002999:Clone Cells; D036101:Cord Blood Stem Cell Transplantation; D005822:Genetic Vectors; D006085:Graft Survival; D006410:Hematopoiesis; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D006801:Humans; D051379:Mice; D016513:Mice, SCID; D041681:NIH 3T3 Cells; D017467:Receptors, Erythropoietin / Q000378:metabolism*; D012190:Retroviridae / Q000235:genetics; D014161:Transduction, Genetic; D019076:Transgenes; D014185:Transplantation, Isogeneic
chemical_list: D018952:Antigens, CD34; D017467:Receptors, Erythropoietin