PLoS ONE: Therapeutic Hemoglobin Levels after Gene Transfer in β-Thalassemia Mice and in Hematopoietic Cells of β-Thalassemia and Sickle Cells Disease Patients
Therapeutic Hemoglobin Levels after Gene Transfer in β-Thalassemia Mice and in Hematopoietic Cells of β-Thalassemia and Sickle Cells Disease Patients
1 Department of Pediatrics, Division of Hematology-Oncology, Children's Blood and Cancer Foundation Laboratories, Weill Cornell Medical College, New York, New York, United States of America, 2 Dipartimento di Biochimica e Biologia Molecolare, Universita' di Ferrara, Ferrara, Italy, 3 Department of Molecular Pharmacology and Chemistry, Memorial Sloan Kettering Cancer Center, New York, New York, United States of America, 4 Red Cell Physiology Laboratory, New York Blood Center, New York, New York, United States of America, 5 Centro della Microcitemia e Anemie Congenite, Ospedali Galliera, Genova, Italy, 6 Department of Pediatrics, Albert Einstein College of Medicine, Children's Hospital at Montefiore, Bronx, New York, United States of America, 7 Department of Medicine, Montefiore Medical Center, Bronx, New York, United States of America, 8 Department of Pharmacology, Langone Medical Center, New York University, New York, New York, United States of America, 9 Department of Hematology, Hadassah–Hebrew University Medical Center, Jerusalem, Israel, 10 Department of Cell and Developmental Biology, Weill Cornell Medical College, New York, New York, United States of America
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.
Citation: Breda L, Casu C, Gardenghi S, Bianchi N, Cartegni L, et al. (2012) Therapeutic Hemoglobin Levels after Gene Transfer in β-Thalassemia Mice and in Hematopoietic Cells of β-Thalassemia and Sickle Cells Disease Patients. PLoS ONE 7(3): e32345. doi:10.1371/journal.pone.0032345
Editor: Christopher B. Doering, Emory University School of Medicine, United States of America
Received: July 13, 2011; Accepted: January 25, 2012; Published: March 27, 2012
Copyright: © 2012 Breda et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by the Cooley's Anemia Foundation (CAF), the Associazione Veneta Lotta alla Talassemia (AVLT; Veneta Association for the Fight Against Thalassemia — Italy) (to L. Breda). This work is supported by grants from the Carlo and Micol Schejola Foundation, the Children's Cancer and Blood Foundation and United States National Institutes of Health grants 5K01DK063992 and NHLBI-5R01HL102449 (to S. Rivella). Additional support was provided by the Clinical and Translational Science Center (CTSC; NIH grant UL1RR024992) and the Carlo and Micol Schejola Foundation (to S. Rivella). RG was funded by Telethon (contract #GGP10124). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: S. Rivella is a consultant for Novartis. In addition, he is a co-inventor for the patents US8058061 B2 C12N 20111115 and US7541179 B2 C12N 20090602. The consulting work and intellectual property of S. Rivella did not affect in any way the design, conduct, or reporting of this research. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials.
* E-mail: str2010@med.cornell.edu
Editor: Christopher B. Doering, Emory University School of Medicine, United States of America
Received: July 13, 2011; Accepted: January 25, 2012; Published: March 27, 2012
Copyright: © 2012 Breda et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
Funding: This work was supported by the Cooley's Anemia Foundation (CAF), the Associazione Veneta Lotta alla Talassemia (AVLT; Veneta Association for the Fight Against Thalassemia — Italy) (to L. Breda). This work is supported by grants from the Carlo and Micol Schejola Foundation, the Children's Cancer and Blood Foundation and United States National Institutes of Health grants 5K01DK063992 and NHLBI-5R01HL102449 (to S. Rivella). Additional support was provided by the Clinical and Translational Science Center (CTSC; NIH grant UL1RR024992) and the Carlo and Micol Schejola Foundation (to S. Rivella). RG was funded by Telethon (contract #GGP10124). The funders had no role in the study design, data collection and analysis, decision to publish, or preparation of the manuscript.
Competing interests: S. Rivella is a consultant for Novartis. In addition, he is a co-inventor for the patents US8058061 B2 C12N 20111115 and US7541179 B2 C12N 20090602. The consulting work and intellectual property of S. Rivella did not affect in any way the design, conduct, or reporting of this research. This does not alter the authors' adherence to all the PLoS ONE policies on sharing data and materials.
* E-mail: str2010@med.cornell.edu
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