A Novel BaEVRless-Pseudotyped γ-Globin Lentiviral Vector Drives High and Stable Fetal Hemoglobin Expression and Improves Thalassemic Erythropoiesis In Vitro .

Autor: Drakopoulou E; 1 Laboratory of Cell and Gene Therapy, Centre for Basic Research, Biomedical Research Foundation of the Academy of Athens (BRFAA), Athens, Greece.; 2 Laboratory of Biology, University of Athens School of Medicine, Athens, Greece., Georgomanoli M; 1 Laboratory of Cell and Gene Therapy, Centre for Basic Research, Biomedical Research Foundation of the Academy of Athens (BRFAA), Athens, Greece.; 2 Laboratory of Biology, University of Athens School of Medicine, Athens, Greece., Lederer CW; 3 Department of Molecular Genetics Thalassemia, The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus.; 4 Cyprus School of Molecular Medicine, Nicosia, Cyprus., Kleanthous M; 3 Department of Molecular Genetics Thalassemia, The Cyprus Institute of Neurology and Genetics, Nicosia, Cyprus.; 4 Cyprus School of Molecular Medicine, Nicosia, Cyprus., Costa C; 5 CIRI-International Center for Infectiology Research, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR 5308, Ecole Normale Supérieure de Lyon, Lyon, France., Bernadin O; 5 CIRI-International Center for Infectiology Research, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR 5308, Ecole Normale Supérieure de Lyon, Lyon, France., Cosset FL; 5 CIRI-International Center for Infectiology Research, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR 5308, Ecole Normale Supérieure de Lyon, Lyon, France., Voskaridou E; 6 Thalassemia and Sickle Cell Disease Centre, Laikon General Hospital, Athens, Greece., Verhoeyen E; 5 CIRI-International Center for Infectiology Research, Inserm, U1111, Université Claude Bernard Lyon 1, CNRS, UMR 5308, Ecole Normale Supérieure de Lyon, Lyon, France.; 7 Inserm, U1065, Centre Méditerranéen de Médecine Moléculaire (C3M), Equipe Contrôle Métabolique des Morts Cellulaires, Nice, France., Papanikolaou E; 1 Laboratory of Cell and Gene Therapy, Centre for Basic Research, Biomedical Research Foundation of the Academy of Athens (BRFAA), Athens, Greece.; 2 Laboratory of Biology, University of Athens School of Medicine, Athens, Greece., Anagnou NP; 1 Laboratory of Cell and Gene Therapy, Centre for Basic Research, Biomedical Research Foundation of the Academy of Athens (BRFAA), Athens, Greece.; 2 Laboratory of Biology, University of Athens School of Medicine, Athens, Greece.
Jazyk: angličtina
Zdroj: Human gene therapy [Hum Gene Ther] 2019 May; Vol. 30 (5), pp. 601-617. Date of Electronic Publication: 2019 Mar 15.
DOI: 10.1089/hum.2018.022
Abstrakt: It has previously been demonstrated that the self-inactivating γ-globin lentiviral vector GGHI can significantly increase fetal hemoglobin (HbF) in erythroid cells from thalassemia patients and thus improve the disease phenotype in vitro. In the present study, the GGHI vector was improved further by incorporating novel enhancer elements and also pseudotyping it with the baboon endogenous virus envelope glycoprotein BaEVRless, which efficiently and specifically targets human CD34 + cells. We evaluated the hypothesis that the newly constructed vector designated as GGHI-mB-3D would increase hCD34 + cell tropism and thus transduction efficiency at low multiplicity of infection, leading to increased transgene expression. High and stable HbF expression was demonstrated in thalassemic cells for the resulting GGHI-mB-3D/BaEVRless vector, exhibiting increased transduction efficiency compared to the original GGHI-mB-3D/VSVG vector, with a concomitant 91% mean HbF increase at a mean vector copy number per cell of 0.86 and a mean transduction efficiency of 56.4%. Transduced populations also exhibited a trend toward late erythroid, orthochromatic differentiation and reduced apoptosis, a further indication of successful gene therapy treatment. Monitoring expression of ATG5 , a key link between autophagy and apoptosis, it was established that this correction correlates with a reduction of enhanced autophagy activation, a typical feature of thalassemic polychromatophilic normoblasts. This work provides novel mechanistic insights into gene therapy-mediated correction of erythropoiesis and demonstrates the beneficial role of BaEVRless envelope glycoprotein compared to VSVG pseudotyping and of the novel GGHI-mB-3D/BaEVRless lentiviral vector for enhanced thalassemia gene therapy.
Databáze: MEDLINE
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