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

Laura Breda, Carla Casu, Sara Gardenghi, Nicoletta Bianchi, Luca Cartegni, Mohandas Narla, Karina Yazdanbakhsh, Marco Musso, Deepa G. Manwani, Jane Little, Lawrence B. Gardner, Dorothy A. Kleinert, Eugenia Prus, Eitan Fibach, Robert W. Grady, Patricia J. Giardina, Roberto Gambari, Stefano Rivella

Research output: Contribution to journalArticle

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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.

Original languageEnglish (US)
Article numbere32345
JournalPLoS One
Volume7
Issue number3
DOIs
StatePublished - Mar 27 2012

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thalassemia
sickle cell anemia
Gene transfer
Thalassemia
Sickle Cell Anemia
Globins
gene transfer
hemoglobin
Hemoglobins
Ankyrins
ankyrins
therapeutics
mice
Genes
clinical trials
cells
Therapeutics
Clinical Trials
bone marrow transplant
erythropoiesis

ASJC Scopus subject areas

  • Agricultural and Biological Sciences(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Medicine(all)

Cite this

Therapeutic hemoglobin levels after gene transfer in β-thalassemia mice and in hematopoietic cells of β-thalassemia and sickle cells disease patients. / Breda, Laura; Casu, Carla; Gardenghi, Sara; Bianchi, Nicoletta; Cartegni, Luca; Narla, Mohandas; Yazdanbakhsh, Karina; Musso, Marco; Manwani, Deepa G.; Little, Jane; Gardner, Lawrence B.; Kleinert, Dorothy A.; Prus, Eugenia; Fibach, Eitan; Grady, Robert W.; Giardina, Patricia J.; Gambari, Roberto; Rivella, Stefano.

In: PLoS One, Vol. 7, No. 3, e32345, 27.03.2012.

Research output: Contribution to journalArticle

Breda, L, Casu, C, Gardenghi, S, Bianchi, N, Cartegni, L, Narla, M, Yazdanbakhsh, K, Musso, M, Manwani, DG, Little, J, Gardner, LB, Kleinert, DA, Prus, E, Fibach, E, Grady, RW, Giardina, PJ, Gambari, R & Rivella, S 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, vol. 7, no. 3, e32345. https://doi.org/10.1371/journal.pone.0032345
Breda, Laura ; Casu, Carla ; Gardenghi, Sara ; Bianchi, Nicoletta ; Cartegni, Luca ; Narla, Mohandas ; Yazdanbakhsh, Karina ; Musso, Marco ; Manwani, Deepa G. ; Little, Jane ; Gardner, Lawrence B. ; Kleinert, Dorothy A. ; Prus, Eugenia ; Fibach, Eitan ; Grady, Robert W. ; Giardina, Patricia J. ; Gambari, Roberto ; Rivella, Stefano. / Therapeutic hemoglobin levels after gene transfer in β-thalassemia mice and in hematopoietic cells of β-thalassemia and sickle cells disease patients. In: PLoS One. 2012 ; Vol. 7, No. 3.
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AU - Narla, Mohandas

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