Ex Vivo Selection of Transduced Hematopoietic Stem Cells for Gene Therapy of β-Hemoglobinopathies

Kanit Bhukhai 1, 2 Edouard de Dreuzy 1, 2 Marie Giorgi 1, 2 Charlotte Colomb 1, 2 Olivier Negre 1, 3, 2 Maria Denaro 3 Beatrix Gillet-Legrand 1, 4, 2 Joëlle Cheuzeville 1, 4, 2 Anaïs Paulard 1, 4, 2 Hélène Trebeden-Negre 5 Suparerk Borwornpinyo 6 Karine Sii-Felice 1, 2 Leila Maouche 1, 2 Julian Down 7 Phillippe Leboulch 1, 2, 8, 9, * Emmanuel Payen 1, 2, *
* Corresponding author
1 IMETI - Institut des Maladies Emergentes et des Thérapies Innovantes
2 UMR E007 - Thérapie génique et contrôle de l'expansion cellulaire
3 Bluebird bio, Inc
4 Bluebird bio France [Fontenay aux Roses]
5 Unité de Thérapie cellulaire [AP-HP Hôpital Pitié-Salpêtrière]
6 Mahidol University [Bangkok, Thailand]
7 Institute for Medical Engineering and Science [Cambridge, MA, USA]
8 Ramathibodi Hospital [Bangkok, Thailand]
9 HMS - Harvard Medical School [Boston]
Abstract : Although gene transfer to hematopoietic stem cells (HSCs) has shown therapeutic efficacy in recent trials for several individuals with inherited disorders, transduction incompleteness of the HSC population remains a hurdle to yield a cure for all patients with reasonably low integrated vector numbers. In previous attempts at HSC selection, massive loss of transduced HSCs, contamination with non-transduced cells, or lack of applicability to large cell populations has rendered the procedures out of reach for human applications. Here, we fused codon-optimized puromycin N-acetyltransferase to herpes simplex virus thymidine kinase. When expressed from a ubiquitous promoter within a complex lentiviral vector comprising the b AT87Q-globin gene, viral titers and therapeutic gene expression were maintained at effective levels. Complete selection and preservation of transduced HSCs were achieved after brief exposure to puromycin in the presence of MDR1 blocking agents, suggesting the procedure's suitability for human clinical applications while affording the additional safety of conditional suicide.
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Submitted on : Tuesday, July 10, 2018 - 11:24:44 PM
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Kanit Bhukhai, Edouard de Dreuzy, Marie Giorgi, Charlotte Colomb, Olivier Negre, et al.. Ex Vivo Selection of Transduced Hematopoietic Stem Cells for Gene Therapy of β-Hemoglobinopathies. Molecular Therapy, Nature Publishing Group, 2018, 26 (2), pp.480 - 495. ⟨10.1016/j.ymthe.2017.10.015⟩. ⟨inserm-01834780⟩

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