Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene Therapy
| العنوان: | Preclinical Evaluation of a Novel Lentiviral Vector Driving Lineage-Specific BCL11A Knockdown for Sickle Cell Gene Therapy |
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| المؤلفون: | Olivier Negre, Swaroopa Guda, Christian Brendel, Chad E. Harris, Martin Bentler, Myriam Armant, Melissa Bonner, Erica B. Esrick, John P. Manis, Helene Trebeden-Negre, Axel Schambach, Alla V. Tsytsykova, Danilo Pellin, Michael Rothe, Lauryn Christiansen, Denise Klatt, David A. Williams, Meaghan McGuinness, Daniela Abriss, Geoff Parsons, Gabor Istvan Veres |
| المصدر: | Molecular Therapy: Methods & Clinical Development, Vol 17, Iss, Pp 589-600 (2020) Molecular Therapy. Methods & Clinical Development |
| بيانات النشر: | Elsevier, 2020. |
| سنة النشر: | 2020 |
| مصطلحات موضوعية: | 0301 basic medicine, fetal hemoglobin, lcsh:QH426-470, Genetic enhancement, Cell, CD34, Biology, Article, Viral vector, Small hairpin RNA, 03 medical and health sciences, 0302 clinical medicine, RNA interference, BCL11A, Fetal hemoglobin, Genetics, medicine, shRNAmiR, lcsh:QH573-671, hemoglobinopathies, Molecular Biology, Gene knockdown, lcsh:Cytology, Sickle cell disease, lentiviral vector, gene therapy, lcsh:Genetics, 030104 developmental biology, medicine.anatomical_structure, HbS, 030220 oncology & carcinogenesis, hemoglobin switch, Cancer research, Molecular Medicine |
| الوصف: | In this work we provide preclinical data to support initiation of a first-in-human trial for sickle cell disease (SCD) using an approach that relies on reversal of the developmental fetal-to-adult hemoglobin switch. Erythroid-specific knockdown of BCL11A via a lentiviral-encoded microRNA-adapted short hairpin RNA (shRNAmiR) leads to reactivation of the gamma-globin gene while simultaneously reducing expression of the pathogenic adult sickle β-globin. We generated a refined lentiviral vector (LVV) BCH-BB694 that was developed to overcome poor vector titers observed in the manufacturing scale-up of the original research-grade LVV. Healthy or sickle cell donor CD34+ cells transduced with Good Manufacturing Practices (GMP)-grade BCH-BB694 LVV achieved high vector copy numbers (VCNs) >5 and gene marking of >80%, resulting in a 3- to 5-fold induction of fetal hemoglobin (HbF) compared with mock-transduced cells without affecting growth, differentiation, and engraftment of gene-modified cells in vitro or in vivo. In vitro immortalization assays, which are designed to measure vector-mediated genotoxicity, showed no increased immortalization compared with mock-transduced cells. Together these data demonstrate that BCH-BB694 LVV is non-toxic and efficacious in preclinical studies, and can be generated at a clinically relevant scale in a GMP setting at high titer to support clinical testing for the treatment of SCD. |
| اللغة: | English |
| تدمد: | 2329-0501 |
| URL الوصول: | https://explore.openaire.eu/search/publication?articleId=doi_dedup___::fb8fc0356964dd1fbe99f38977e8a66a http://www.sciencedirect.com/science/article/pii/S2329050120300450 |
| حقوق: | OPEN |
| رقم الأكسشن: | edsair.doi.dedup.....fb8fc0356964dd1fbe99f38977e8a66a |
| قاعدة البيانات: | OpenAIRE |
| تدمد: | 23290501 |
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