Therapeutic hemoglobin levels after gene transfer in β-thalassemia mice and in hematopoietic cells of β-thalassemia and sickle cells disease patients Journal Article


Authors: Breda, L.; Casu, C.; Gardenghi, S.; Bianchi, N.; Cartegni, L.; Narla, M.; Yazdanbakhsh, K.; Musso, M.; Manwani, D.; Little, J.; Gardner, L. B.; Kleinert, D. A.; Prus, E.; Fibach, E.; Grady, R. W.; Giardina, P. J.; Gambari, R.; Rivella, S.
Article Title: Therapeutic hemoglobin levels after gene transfer in β-thalassemia mice and in hematopoietic cells of β-thalassemia and sickle cells disease patients
Abstract: Preclinical and clinical studies demonstrate the feasibility of treating β-thalassemia and Sickle Cell Disease (SCD) by lentiviral-mediated transfer of the human β-globin gene. However, previous studies have not addressed whether the ability of lentiviral vectors to increase hemoglobin synthesis might vary in different patients. We generated lentiviral vectors carrying the human β-globin gene with and without an ankyrin insulator and compared their ability to induce hemoglobin synthesis in vitro and in thalassemic mice. We found that insertion of an ankyrin insulator leads to higher, potentially therapeutic levels of human β-globin through a novel mechanism that links the rate of transcription of the transgenic β-globin mRNA during erythroid differentiation with polysomal binding and efficient translation, as reported here for the first time. We also established a preclinical assay to test the ability of this novel vector to synthesize adult hemoglobin in erythroid precursors and in CD34 + cells isolated from patients affected by β-thalassemia and SCD. Among the thalassemic patients, we identified a subset of specimens in which hemoglobin production can be achieved using fewer copies of the vector integrated than in others. In SCD specimens the treatment with AnkT9W ameliorates erythropoiesis by increasing adult hemoglobin (Hb A) and concurrently reducing the sickling tetramer (Hb S). Our results suggest two major findings. First, we discovered that for the purpose of expressing the β-globin gene the ankyrin element is particularly suitable. Second, our analysis of a large group of specimens from β-thalassemic and SCD patients indicates that clinical trials could benefit from a simple test to predict the relationship between the number of vector copies integrated and the total amount of hemoglobin produced in the erythroid cells of prospective patients. This approach would provide vital information to select the best candidates for these clinical trials, before patients undergo myeloablation and bone marrow transplant. © 2012 Breda et al.
Keywords: adult; controlled study; gene mutation; human cell; genetics; mutation; nonhuman; molecular genetics; methodology; mouse; animal; metabolism; animals; mice; gene; cells, cultured; mus; cd34 antigen; gene expression; erythroid precursor cell; erythropoiesis; spleen; erythroid precursor cells; animal experiment; animal model; hemoglobin; hemoglobin blood level; hematopoietic stem cell transplantation; cell differentiation; in vitro study; cell line, tumor; gene transfer; viral gene delivery system; gene vector; genetic vectors; chimera; blood; molecular sequence data; cell culture; messenger rna; 5' untranslated region; nucleotide sequence; splenomegaly; tumor cell line; cell isolation; gene therapy; lentivirus; iron; lentivirus vector; beta thalassemia; erythroid cell; hematopoietic cell; transgene; beta-globins; beta-thalassemia; rna translation; base sequence; erythrocyte; 3' untranslated region; gene dosage; gene insertion; hermes antigen; bone marrow transplantation; antigens, cd34; cd34 selection; hemoglobins; cell strain 3t3; nih 3t3 cells; rna transcription; reticulocyte count; lentivirinae; hematocrit; erythrocyte structure; gene transfer techniques; beta globin; sickle cell anemia; spleen size; anemia, sickle cell; hemoglobin f; hemoglobin s; insulator element; insulator elements; hemoglobin synthesis; alpha globin; ankyrins; ankyrin; cd71 antigen; hemoglobin a; ankyrin insulator element; beta globin gene; iron liver level; liver level; red cell distribution width; spleen iron
Journal Title: PLoS ONE
Volume: 7
Issue: 3
ISSN: 1932-6203
Publisher: Public Library of Science  
Date Published: 2012-01-01
Start Page: e32345
Language: English
DOI: 10.1371/journal.pone.0032345
PROVIDER: scopus
PMCID: PMC3314006
PUBMED: 22479321
DOI/URL:
Notes: --- - "Export Date: 1 May 2012" - "Source: Scopus"
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