title: Plerixafor+G-CSF-mobilized CD34+ cells represent an optimal graft source for thalassemia gene therapy.
abstract: Globin gene therapy requires abundant numbers of highly engraftable, autologous hematopoietic stem cells expressing curative levels of β-globin on differentiation. In this study, CD34+ cells from 31 thalassemic patients mobilized with hydroxyurea+granulocyte colony-stimulating factor (G-CSF), G-CSF, Plerixafor, or Plerixafor+G-CSF were transduced with the TNS9.3.55 β-globin lentivector and compared for transducibility and globin expression in vitro, as well as engraftment potential in a xenogeneic model after partial myeloablation. Transduction efficiency and vector copy number (VCN) averaged 48.4 ± 2.8% and 1.91 ± 0.04, respectively, whereas expression approximated the one-copy normal β-globin output. Plerixafor+G-CSF cells produced the highest β-globin expression/VCN. Long-term multilineage engraftment and persistent VCN and vector expression was encountered in all xenografted groups, with Plerixafor+G-CSF-mobilized cells achieving superior short-term engraftment rates, with similar numbers of CD34+ cells transplanted. Overall, Plerixafor+G-CSF not only allows high CD34+ cell yields but also provides increased β-globin expression/VCN and enhanced early human chimerism under nonmyeloablative conditions, thus representing an optimal graft for thalassemia gene therapy.
authors.affiliation: Gene and Cell Therapy Center, Hematology Department-Bone Marrow Transplantation Unit, George Papanicolaou Hospital, Thessaloniki, Greece; School of Medicine, Democritus University of Thrace, Alexandroupolis, Greece;
authors.forename: Garyfalia
authors.initials: G
authors.lastname: Karponi
authors.affiliation: Gene and Cell Therapy Center, Hematology Department-Bone Marrow Transplantation Unit, George Papanicolaou Hospital, Thessaloniki, Greece;
authors.forename: Nikoletta
authors.initials: N
authors.lastname: Psatha
authors.affiliation: Molecular Genetics Thalassaemia Department, The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus;
authors.forename: Carsten Werner
authors.initials: CW
authors.lastname: Lederer
authors.affiliation: Clinical Research Division, Fred Hutchinson Cancer Research Center, Seattle, WA; Department of Medicine and Markey Molecular Medicine Center, University of Washington, Seattle, WA;
authors.forename: Jennifer Eileen
authors.initials: JE
authors.lastname: Adair
authors.affiliation: Gene and Cell Therapy Center, Hematology Department-Bone Marrow Transplantation Unit, George Papanicolaou Hospital, Thessaloniki, Greece;
authors.forename: Fani
authors.initials: F
authors.lastname: Zervou
authors.affiliation: Gene and Cell Therapy Center, Hematology Department-Bone Marrow Transplantation Unit, George Papanicolaou Hospital, Thessaloniki, Greece;
authors.forename: Nikolaos
authors.initials: N
authors.lastname: Zogas
authors.affiliation: Molecular Genetics Thalassaemia Department, The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus;
authors.forename: Marina
authors.initials: M
authors.lastname: Kleanthous
authors.affiliation: School of Medicine, Democritus University of Thrace, Alexandroupolis, Greece;
authors.forename: Constantinos
authors.initials: C
authors.lastname: Tsatalas
authors.affiliation: Gene and Cell Therapy Center, Hematology Department-Bone Marrow Transplantation Unit, George Papanicolaou Hospital, Thessaloniki, Greece;
authors.forename: Achilles
authors.initials: A
authors.lastname: Anagnostopoulos
authors.affiliation: Center for Cell Engineering, Memorial Sloan-Kettering Cancer Center, New York, NY; and.
authors.forename: Michel
authors.initials: M
authors.lastname: Sadelain
authors.affiliation: Center for Cell Engineering, Memorial Sloan-Kettering Cancer Center, New York, NY; and Cell Therapy and Cell Engineering Facility, Memorial Sloan-Kettering Cancer Center, New York, NY.
authors.forename: Isabelle
authors.initials: I
authors.lastname: Rivière
authors.affiliation: Department of Medicine and Markey Molecular Medicine Center, University of Washington, Seattle, WA;
authors.forename: George
authors.initials: G
authors.lastname: Stamatoyannopoulos
authors.affiliation: Gene and Cell Therapy Center, Hematology Department-Bone Marrow Transplantation Unit, George Papanicolaou Hospital, Thessaloniki, Greece;
authors.forename: Evangelia
authors.initials: E
authors.lastname: Yannaki
mesh_terms: D000818:Animals; D018952:Antigens, CD34 / Q000378:metabolism; D001596:Benzylamines; D000080027:Cyclams; D018628:Gene Dosage; D015870:Gene Expression; D015316:Genetic Therapy / Q000379:methods*; D005822:Genetic Vectors; D016179:Granulocyte Colony-Stimulating Factor / Q000008:administration & dosage*; D019650:Hematopoietic Stem Cell Mobilization / Q000379:methods*; D018380:Hematopoietic Stem Cell Transplantation; D006412:Hematopoietic Stem Cells / Q000378:metabolism; D006571:Heterocyclic Compounds / Q000008:administration & dosage; D064593:Heterografts; D006801:Humans; D051379:Mice; D018345:Mice, Knockout; D014182:Transplantation, Autologous; D055544:beta-Globins / Q000235:genetics; D017086:beta-Thalassemia / Q000235:genetics / Q000628:therapy*
chemical_list: D018952:Antigens, CD34; D001596:Benzylamines; D000080027:Cyclams; D006571:Heterocyclic Compounds; D055544:beta-Globins; D016179:Granulocyte Colony-Stimulating Factor; C088327:plerixafor