title: Proliferation deficiency of multipotent hematopoietic progenitors in ribosomal protein S19 (RPS19)-deficient diamond-Blackfan anemia improves following RPS19 gene transfer.
abstract: Diamond-Blackfan anemia (DBA) is a congenital bone marrow failure syndrome characterized by a specific deficiency in erythroid progenitors. Since some patients with DBA develop a reduction in thrombocytes and granulocytes with age, we asked whether multipotent hematopoietic progenitors from DBA patients had normal proliferative capacity in liquid expansion cultures. CD34(+) cells derived from DBA patients showed deficient proliferation in liquid culture containing IL-3, IL-6, and SCF. Single CD34(+) CD38(-) cells from DBA patients exhibited deficient proliferation recruitment in a limiting dilution assay containing IL-3, IL-6, SCF, Tpo, FL, and G-CSF or containing IL-3, IL-6, and SCF. Our findings suggest that the underlying hematopoietic defect in DBA may not be limited to the erythroid lineage. Since a fraction of DBA patients have a deficiency in ribosomal protein S19 (RPS19), we constructed lentiviral vectors containing the RPS19 gene for overexpression in hematopoietic progenitors from RPS19-deficient DBA patients. Enforced expression of the RPS19 transgene improved the proliferation of CD34(+) cells from DBA patients with RPS19 mutation. Similarly, enforced expression of RPS19 improved erythroid development of RPS19-deficient hematopoietic progenitors as determined by colony assays and erythroid differentiation cultures. These findings suggest that gene therapy for RPS19-deficient DBA is feasible.
authors.affiliation: Molecular Medicine and Gene Therapy, 221 84 Lund, Sweden.
authors.forename: Isao
authors.initials: I
authors.lastname: Hamaguchi
authors.forename: Johan
authors.initials: J
authors.lastname: Flygare
authors.forename: Hiroshi
authors.initials: H
authors.lastname: Nishiura
authors.forename: Ann C M
authors.initials: AC
authors.lastname: Brun
authors.forename: Andreas
authors.initials: A
authors.lastname: Ooka
authors.forename: Thomas
authors.initials: T
authors.lastname: Kiefer
authors.forename: Zhi
authors.initials: Z
authors.lastname: Ma
authors.forename: Niklas
authors.initials: N
authors.lastname: Dahl
authors.forename: Johan
authors.initials: J
authors.lastname: Richter
authors.forename: Stefan
authors.initials: S
authors.lastname: Karlsson
mesh_terms: D029503:Anemia, Diamond-Blackfan / Q000378:metabolism / Q000628:therapy*; D015703:Antigens, CD / Q000032:analysis; D001853:Bone Marrow / Q000473:pathology; D016022:Case-Control Studies; D002455:Cell Division / Q000502:physiology; D002460:Cell Line; D003114:Colony-Forming Units Assay; D016207:Cytokines / Q000378:metabolism; D005786:Gene Expression Regulation / Q000502:physiology*; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors; D006412:Hematopoietic Stem Cells / Q000378:metabolism* / Q000473:pathology; D006801:Humans; D016086:Lentivirus / Q000235:genetics; D012269:Ribosomal Proteins / Q000172:deficiency / Q000235:genetics*; D014162:Transfection
chemical_list: D015703:Antigens, CD; D016207:Cytokines; D012269:Ribosomal Proteins; C040343:ribosomal protein S19