title: Gene therapy of Diamond Blackfan anemia CD34(+) cells leads to improved erythroid development and engraftment following transplantation.
abstract: OBJECTIVE
  Diamond-Blackfan anemia (DBA) is a rare congenital hypoplastic anemia caused by mutations in ribosomal protein (RP) genes. Our aim is to develop gene therapy for DBA patients with mutations in RPS19. We previously demonstrated that RPS19 gene transfer partially corrects erythroid development in vitro. In this study, we asked if RPS19 gene transfer corrects erythroid development in unsorted cells transplanted to immunodeficient mice and if the RPS19-corrected fraction has a proliferative advantage after transplantation. We further determined if high level of RPS19 expression is required for correction.
  
  
  METHODS
  Mobilized peripheral blood CD34(+) cells were transduced by oncoretroviral vector particles pseudotyped with the feline endogenous retrovirus envelope. Vectors containing two different promoters with different RPS19 transgene expression levels were compared. Transduced cells were transplanted to immunocompromised nonobese diabetic/severe combined immunodeficient-beta2 microglobulin null mice in order to assess therapeutic effects of RPS19 gene transfer in vivo.
  
  
  RESULTS
  We show that correction of erythroid development requires high RPS19 expression. The corrected fraction of unselected DBA cells have a survival advantage in vivo, suggesting that successful gene therapy may only require correction of a fraction of the patient cells.
  
  
  CONCLUSIONS
  Our findings are fundamental for development of clinical gene therapy for DBA because they demonstrate increased engraftment of RPS19-transduced cells without selection of gene-corrected cells prior to transplantation, an essential prelude to studying long-term therapeutic effects in emerging animal models for DBA.
authors: Department of Molecular Medicine and Gene Therapy, Lund Stem Cell Center, Lund University Hospital, Lund, Sweden. flygare@wi.mit.edu
authors: Johan
authors: J
authors: Flygare
authors: Karin
authors: K
authors: Olsson
authors: Johan
authors: J
authors: Richter
authors: Stefan
authors: S
authors: Karlsson
mesh_terms: D000328:Adult; D029503:Anemia, Diamond-Blackfan / Q000097:blood / Q000628:therapy*; D018952:Antigens, CD34 / Q000032:analysis; D004920:Erythropoiesis; D005260:Female; D015316:Genetic Therapy; D018380:Hematopoietic Stem Cell Transplantation; D006801:Humans; D012269:Ribosomal Proteins / Q000235:genetics*
chemical_list: D018952:Antigens, CD34; D012269:Ribosomal Proteins; C040343:ribosomal protein S19