Proliferation deficiency of multipotent hematopoietic progenitors in ribosomal protein S19 (RPS19)-deficient diamond–Blackfan anemia improves following RPS19 gene transfer

• Published in: Molecular therapy Vol. 7; no. 5; pp. 613 - 622
• Main Authors: Hamaguchi, Isao, Flygare, Johan, Nishiura, Hiroshi, Brun, Ann C.M, Ooka, Andreas, Kiefer, Thomas, Ma, Zhi, Dahl, Niklas, Richter, Johan, Karlsson, Stefan
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01-05-2003; Elsevier Limited
• Subjects: Anemia; Anemia, Diamond-Blackfan - metabolism; Anemia, Diamond-Blackfan - therapy; Antigens, CD - analysis; Basic Medicine; Bone marrow; Bone Marrow - pathology; Case-Control Studies; Cell Division - physiology; Cell Line; Colony-Forming Units Assay; Congenital diseases; Cytokines - metabolism; Diamond-Blackfan anemia; Gene Expression Regulation - physiology; Gene therapy; Gene Transfer Techniques; Genetic Therapy; Genetic Vectors; hematopoietic progenitors; Hematopoietic Stem Cells - metabolism; Hematopoietic Stem Cells - pathology; Humans; lentiviral vectors; Lentivirus - genetics; Medical and Health Sciences; Medical Genetics; Medicin och hälsovetenskap; Medicinsk genetik; Medicinska och farmaceutiska grundvetenskaper; Mutation; Neutropenia; Patients; Proteins; Ribosomal Proteins - deficiency; Ribosomal Proteins - genetics; Thrombocytopenia; Transfection; Vectors (Biology); Sweden; gene therapy; hematopoietic progenitors; lentiviral vectors; Diamond–Blackfan anemia
• ISSN: 1525-0016; 1525-0024; 1525-0024
• DOI: 10.1016/S1525-0016(03)00091-1

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.