title: Potentially therapeutic levels of anti-sickling globin gene expression following lentivirus-mediated gene transfer in sickle cell disease bone marrow CD34+ cells.
abstract: Sickle cell disease (SCD) can be cured by allogeneic hematopoietic stem cell transplant. However, this is only possible when a matched donor is available, making the development of gene therapy using autologous hematopoietic stem cells a highly desirable alternative. We used a culture model of human erythropoiesis to directly compare two insulated, self-inactivating, and erythroid-specific lentiviral vectors, encoding for γ-globin (V5m3-400) or a modified β-globin (βAS3-FB) for production of antisickling hemoglobin (Hb) and correction of red cell deformability after deoxygenation. Bone marrow CD34+ cells from three SCD patients were transduced using V5m3-400 or βAS3-FB and compared with mock-transduced SCD or healthy donor CD34+ cells. Lentiviral transduction did not impair cell growth or differentiation, as gauged by proliferation and acquisition of erythroid markers. Vector copy number averaged approximately one copy per cell, and corrective globin mRNA levels were increased more than sevenfold over mock-transduced controls. Erythroblasts derived from healthy donor and mock-transduced SCD cells produced a low level of fetal Hb that was increased to 23.6 ± 4.1% per vector copy for cells transduced with V5m3-400. Equivalent levels of modified normal adult Hb of 17.6 ± 3.8% per vector copy were detected for SCD cells transduced with βAS3-FB. These levels of antisickling Hb production were sufficient to reduce sickling of terminal-stage red blood cells upon deoxygenation. We concluded that the achieved levels of fetal Hb and modified normal adult Hb would likely prove therapeutic to SCD patients who lack matched donors.
authors: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA.
authors: Fabrizia
authors: F
authors: Urbinati
authors: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN, USA.
authors: Phillip W
authors: PW
authors: Hargrove
authors: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA.
authors: Sabine
authors: S
authors: Geiger
authors: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA.
authors: Zulema
authors: Z
authors: Romero
authors: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA.
authors: Jennifer
authors: J
authors: Wherley
authors: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA.
authors: Michael L
authors: ML
authors: Kaufman
authors: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA.
authors: Roger P
authors: RP
authors: Hollis
authors: Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, Springfield, IL, USA.
authors: Christopher B
authors: CB
authors: Chambers
authors: Department of Hematology, St. Jude Children's Research Hospital, Memphis, TN, USA.
authors: Derek A
authors: DA
authors: Persons
authors: Department of Microbiology, Immunology and Molecular Genetics, University of California, Los Angeles, CA, USA.
authors: Donald B
authors: DB
authors: Kohn
authors: Department of Medical Microbiology, Immunology and Cell Biology, Southern Illinois University School of Medicine, Springfield, IL, USA. Electronic address: awilber@siumed.edu.
authors: Andrew
authors: A
authors: Wilber
mesh_terms: D000755:Anemia, Sickle Cell / Q000235:genetics / Q000628:therapy; D018952:Antigens, CD34 / Q000378:metabolism; D001854:Bone Marrow Cells / Q000378:metabolism*; D005319:Fetal Hemoglobin / Q000235:genetics; D005434:Flow Cytometry; D015870:Gene Expression; D018014:Gene Transfer Techniques; D015316:Genetic Therapy / Q000379:methods; D005822:Genetic Vectors / Q000235:genetics; D006454:Hemoglobins / Q000235:genetics; D006801:Humans; D016086:Lentivirus / Q000235:genetics*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D055544:beta-Globins / Q000235:genetics* / Q000378:metabolism; D055545:gamma-Globins / Q000235:genetics* / Q000378:metabolism
chemical_list: D018952:Antigens, CD34; D006454:Hemoglobins; D055544:beta-Globins; D055545:gamma-Globins; D005319:Fetal Hemoglobin