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Therapeutic haemoglobin synthesis in β-thalassaemic mice expressing lentivirus-encoded human β-globin

Abstract

The stable introduction of a functional β-globin gene in haematopoietic stem cells could be a powerful approach to treat β-thalassaemia1 and sickle-cell disease2. Genetic approaches aiming to increase normal β-globin expression in the progeny of autologous haematopoietic stem cells3 might circumvent the limitations and risks of allogeneic cell transplants4. However, low-level expression, position effects and transcriptional silencing hampered the effectiveness of viral transduction of the human β-globin gene when it was linked to minimal regulatory sequences5. Here we show that the use of recombinant lentiviruses enables efficient transfer and faithful integration of the human β-globin gene together with large segments of its locus control region. In long-term recipients of unselected transduced bone marrow cells, tetramers of two murine α-globin and two human βA-globin molecules account for up to 13% of total haemoglobin in mature red cells of normal mice. In β-thalassaemic heterozygous mice higher percentages are obtained (17% to 24%), which are sufficient to ameliorate anaemia and red cell morphology. Such levels should be of therapeutic benefit in patients with severe defects in haemoglobin production.

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Figure 1: Structure and stability of recombinant lentiviral vectors.
Figure 2: Increased mean β-globin expression in MEL cells and proportion of clones expressing detectable human β-globin with the TNS9 vector.
Figure 3: Long-term stability of vector copy number, human β-globin RNA levels and haemoglobin tetramers in peripheral blood of bone marrow chimaeras.
Figure 4: Integration of lentiviral vectors into hematopoietic stem cells.
Figure 5: Correction of anisocytosis and poikilocytosis in bone marrow chimaeras reconstituted with TNS9-transduced Hbbth3/+ bone marrow cells.
Figure 6: Amelioration of haematological parameters in bone marrow chimaeras reconstituted with TNS9-transduced Hbbth3/+ bone marrow cells.

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Acknowledgements

We thank M. Trudel for helpful discussion; C. Tan and H. Beauchemin for technical assistance and I. Rivière for reviewing the manuscript. This work was supported by grants from the NHLBI, the NCI, the Cancer Research Institute, the Cooley's Anaemia Foundation, DeWitt-Wallace Fund and the McDonnell Foundation Scholars Award.

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Correspondence to Michel Sadelain.

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May, C., Rivella, S., Callegari, J. et al. Therapeutic haemoglobin synthesis in β-thalassaemic mice expressing lentivirus-encoded human β-globin. Nature 406, 82–86 (2000). https://doi.org/10.1038/35017565

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