title: Therapeutic levels of fetal hemoglobin in erythroid progeny of β-thalassemic CD34+ cells after lentiviral vector-mediated gene transfer.
abstract: β-Thalassemia major results from severely reduced or absent expression of the β-chain of adult hemoglobin (α₂β₂;HbA). Increased levels of fetal hemoglobin (α₂γ₂;HbF), such as occurs with hereditary persistence of HbF, ameliorate the severity of β-thalassemia, raising the potential for genetic therapy directed at enhancing HbF. We used an in vitro model of human erythropoiesis to assay for enhanced production of HbF after gene delivery into CD34(+) cells obtained from mobilized peripheral blood of normal adults or steady-state bone marrow from patients with β-thalassemia major. Lentiviral vectors encoding (1) a human γ-globin gene with or without an insulator, (2) a synthetic zinc-finger transcription factor designed to interact with the γ-globin gene promoters, or (3) a short-hairpin RNA targeting the γ-globin gene repressor, BCL11A, were tested. Erythroid progeny of normal CD34(+) cells demonstrated levels of HbF up to 21% per vector copy. For β-thalassemic CD34(+) cells, similar gene transfer efficiencies achieved HbF production ranging from 45% to 60%, resulting in up to a 3-fold increase in the total cellular Hb content. These observations suggest that both lentiviral-mediated γ-globin gene addition and genetic reactivation of endogenous γ-globin genes have potential to provide therapeutic HbF levels to patients with β-globin deficiency.
authors.affiliation: Department of Surgery, Southern Illinois University School of Medicine, Springfield, IL, USA.
authors.forename: Andrew
authors.initials: A
authors.lastname: Wilber
authors.forename: Phillip W
authors.initials: PW
authors.lastname: Hargrove
authors.forename: Yoon-Sang
authors.initials: YS
authors.lastname: Kim
authors.forename: Janice M
authors.initials: JM
authors.lastname: Riberdy
authors.forename: Vijay G
authors.initials: VG
authors.lastname: Sankaran
authors.forename: Eleni
authors.initials: E
authors.lastname: Papanikolaou
authors.forename: Maria
authors.initials: M
authors.lastname: Georgomanoli
authors.forename: Nicholas P
authors.initials: NP
authors.lastname: Anagnou
authors.forename: Stuart H
authors.initials: SH
authors.lastname: Orkin
authors.forename: Arthur W
authors.initials: AW
authors.lastname: Nienhuis
authors.forename: Derek A
authors.initials: DA
authors.lastname: Persons
mesh_terms: D018952:Antigens, CD34 / Q000378:metabolism; D015139:Blotting, Southern; D015153:Blotting, Western; D002469:Cell Separation; D015672:Erythroid Precursor Cells / Q000378:metabolism*; D004920:Erythropoiesis / Q000502:physiology; D005319:Fetal Hemoglobin / Q000096:biosynthesis* / Q000235:genetics; D005434:Flow Cytometry; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D016133:Polymerase Chain Reaction; D017086:beta-Thalassemia / Q000628:therapy*; D055545:gamma-Globins / Q000235:genetics*
chemical_list: D018952:Antigens, CD34; D055545:gamma-Globins; D005319:Fetal Hemoglobin