Combination of lentiviral and genome editing technologies for the treatment of sickle cell disease.

Autor:	Ramadier S; Laboratory of Chromatin and Gene Regulation during Development, Imagine Institute, INSERM UMR1163, 75015 Paris, France; Université de Paris, 75015 Paris, France; Phasics, Bâtiment Explorer, Espace Technologique, Route de l'Orme des Merisiers, 91190 St. Aubin, France., Chalumeau A; Laboratory of Chromatin and Gene Regulation during Development, Imagine Institute, INSERM UMR1163, 75015 Paris, France; Université de Paris, 75015 Paris, France., Felix T; Laboratory of Chromatin and Gene Regulation during Development, Imagine Institute, INSERM UMR1163, 75015 Paris, France; Université de Paris, 75015 Paris, France., Othman N; Phasics, Bâtiment Explorer, Espace Technologique, Route de l'Orme des Merisiers, 91190 St. Aubin, France., Aknoun S; Phasics, Bâtiment Explorer, Espace Technologique, Route de l'Orme des Merisiers, 91190 St. Aubin, France., Casini A; CIBIO, University of Trento, 38100 Trento, Italy., Maule G; CIBIO, University of Trento, 38100 Trento, Italy., Masson C; Paris-Descartes Bioinformatics Platform, Imagine Institute, 75015 Paris, France., De Cian A; INSERM U1154, CNRS UMR7196, Museum National d'Histoire Naturelle, 75015 Paris, France., Frati G; Laboratory of Chromatin and Gene Regulation during Development, Imagine Institute, INSERM UMR1163, 75015 Paris, France; Université de Paris, 75015 Paris, France., Brusson M; Laboratory of Chromatin and Gene Regulation during Development, Imagine Institute, INSERM UMR1163, 75015 Paris, France; Université de Paris, 75015 Paris, France., Concordet JP; INSERM U1154, CNRS UMR7196, Museum National d'Histoire Naturelle, 75015 Paris, France., Cavazzana M; Université de Paris, 75015 Paris, France; Imagine Institute, 75015 Paris, France; Biotherapy Department and Clinical Investigation Center, Assistance Publique Hôpitaux de Paris, INSERM, 75015 Paris, France., Cereseto A; CIBIO, University of Trento, 38100 Trento, Italy., El Nemer W; Etablissement Français du Sang PACA-Corse, Marseille, France; Aix Marseille Université, EFS, CNRS, ADES, 'Biologie des Groupes Sanguins,' 13000 Marseille, France; Laboratoire d'Excellence GR-Ex, Paris, France., Amendola M; Genethon, INSERM UMR951, 91000 Evry, France., Wattellier B; Phasics, Bâtiment Explorer, Espace Technologique, Route de l'Orme des Merisiers, 91190 St. Aubin, France., Meneghini V; Laboratory of Chromatin and Gene Regulation during Development, Imagine Institute, INSERM UMR1163, 75015 Paris, France; Université de Paris, 75015 Paris, France. Electronic address: meneghini.vasco@hsr.it., Miccio A; Laboratory of Chromatin and Gene Regulation during Development, Imagine Institute, INSERM UMR1163, 75015 Paris, France; Université de Paris, 75015 Paris, France. Electronic address: annarita.miccio@institutimagine.org.
Jazyk:	angličtina
Zdroj:	Molecular therapy : the journal of the American Society of Gene Therapy [Mol Ther] 2022 Jan 05; Vol. 30 (1), pp. 145-163. Date of Electronic Publication: 2021 Aug 19.
DOI:	10.1016/j.ymthe.2021.08.019
Abstrakt:	Sickle cell disease (SCD) is caused by a mutation in the β-globin gene leading to polymerization of the sickle hemoglobin (HbS) and deformation of red blood cells. Autologous transplantation of hematopoietic stem/progenitor cells (HSPCs) genetically modified using lentiviral vectors (LVs) to express an anti-sickling β-globin leads to some clinical benefit in SCD patients, but it requires high-level transgene expression (i.e., high vector copy number [VCN]) to counteract HbS polymerization. Here, we developed therapeutic approaches combining LV-based gene addition and CRISPR-Cas9 strategies aimed to either knock down the sickle β-globin and increase the incorporation of an anti-sickling globin (AS3) in hemoglobin tetramers, or to induce the expression of anti-sickling fetal γ-globins. HSPCs from SCD patients were transduced with LVs expressing AS3 and a guide RNA either targeting the endogenous β-globin gene or regions involved in fetal hemoglobin silencing. Transfection of transduced cells with Cas9 protein resulted in high editing efficiency, elevated levels of anti-sickling hemoglobins, and rescue of the SCD phenotype at a significantly lower VCN compared to the conventional LV-based approach. This versatile platform can improve the efficacy of current gene addition approaches by combining different therapeutic strategies, thus reducing the vector amount required to achieve a therapeutic VCN and the associated genotoxicity risk. Competing Interests: Declaration of interests V.M. and A.M. are the inventors of two patents describing this gene addition/genome editing platform (WO/2018/220211, “Viral vectors combining gene therapy and genome editing approaches for gene therapy of genetic disorders”; WO/2018/220210, “Recombinant lentiviral vector for stem cell-based gene therapy of sickle cell disorder). The remaining authors declare no competing interests. (Copyright © 2021. Published by Elsevier Inc.)
Databáze:	MEDLINE
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