An Optimized Lentiviral Vector Efficiently Corrects the Human Sickle Cell Disease Phenotype

Leslie Weber; Valentina Poletti; Elisa Magrin; Chiara Antoniani; Samia Martin; Charles Bayard; Hanem Sadek; Tristan Felix; Vasco Meneghini; Michael N Antoniou; Wassim El-Nemer; Fulvio Mavilio; Marina Cavazzana; Isabelle Andre-Schmutz; Annarita Miccio

doi:10.1016/j.omtm.2018.07.012

An Optimized Lentiviral Vector Efficiently Corrects the Human Sickle Cell Disease Phenotype

Mol Ther Methods Clin Dev. 2018 Aug 4:10:268-280. doi: 10.1016/j.omtm.2018.07.012. eCollection 2018 Sep 21.

Authors

Leslie Weber^{1

2}, Valentina Poletti³, Elisa Magrin⁴, Chiara Antoniani^{5

6}, Samia Martin³, Charles Bayard¹, Hanem Sadek¹, Tristan Felix^{5

6}, Vasco Meneghini^{5

6}, Michael N Antoniou⁷, Wassim El-Nemer^{8

9

10}, Fulvio Mavilio^{5

11}, Marina Cavazzana^{1

4

5}, Isabelle Andre-Schmutz^{1

5}, Annarita Miccio^{3

5

6}

Affiliations

¹ Laboratory of Human Lymphohematopoiesis, INSERM UMR_S1163, 75015 Paris, France.
² Paris Diderot University - Sorbonne Paris Cité, 75015 Paris, France.
³ Genethon, INSERM UMR951, 91000 Evry, France.
⁴ Biotherapy Department, Necker Children's Hospital, Assistance Publique-Hôpitaux de Paris, 75015 Paris, France.
⁵ Paris Descartes-Sorbonne Paris Cité University, Imagine Institute, 75015 Paris, France.
⁶ Laboratory of chromatin and gene regulation during development, INSERM UMR_S1163, 75015 Paris, France.
⁷ King's College London, WC2R 2LS London, England.
⁸ Biologie Intégrée du Globule Rouge, INSERM UMR_S1134, Paris Diderot University, Sorbonne Paris Cité, Université de la Réunion, Université des Antilles, 75015 Paris, France.
⁹ Institut National de la Transfusion Sanguine, 75015 Paris, France.
¹⁰ Laboratoire d'Excellence GR-Ex, 75015 Paris, France.
¹¹ Department of Life Sciences, University of Modena and Reggio Emilia, 41125 Modena, Italy.

Abstract

Autologous transplantation of hematopoietic stem cells transduced with a lentiviral vector (LV) expressing an anti-sickling HBB variant is a potential treatment for sickle cell disease (SCD). With a clinical trial as our ultimate goal, we generated LV constructs containing an anti-sickling HBB transgene (HBBAS3), a minimal HBB promoter, and different combinations of DNase I hypersensitive sites (HSs) from the locus control region (LCR). Hematopoietic stem progenitor cells (HSPCs) from SCD patients were transduced with LVs containing either HS2 and HS3 (β-AS3) or HS2, HS3, and HS4 (β-AS3 HS4). The inclusion of the HS4 element drastically reduced vector titer and infectivity in HSPCs, with negligible improvement of transgene expression. Conversely, the LV containing only HS2 and HS3 was able to efficiently transduce SCD bone marrow and Plerixafor-mobilized HSPCs, with anti-sickling HBB representing up to ∼60% of the total HBB-like chains. The expression of the anti-sickling HBB and the reduced incorporation of the β^S-chain in hemoglobin tetramers allowed up to 50% reduction in the frequency of RBC sickling under hypoxic conditions. Together, these results demonstrate the ability of a high-titer LV to express elevated levels of a potent anti-sickling HBB transgene ameliorating the SCD cell phenotype.

Keywords: gene therapy; lentiviral vectors; sickle cell disease.