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Optimization of CRISPR/Cas9 Delivery to Human Hematopoietic Stem and Progenitor Cells for Therapeutic Genomic Rearrangements

التفاصيل البيبلوغرافية
العنوان: Optimization of CRISPR/Cas9 Delivery to Human Hematopoietic Stem and Progenitor Cells for Therapeutic Genomic Rearrangements
المؤلفون: Cécile Masson, Mario Amendola, Annarita Miccio, Annalisa Lattanzi, Ciaran M. Lee, Chiara Antoniani, Sophie Ramadier, Gang Bao, Olivier Alibeu, Vasco Meneghini, Tristan Felix, Matthew H. Porteus, Giulia Pavani, Fulvio Mavilio, Fatima Amor
المساهمون: Approches génétiques intégrées et nouvelles thérapies pour les maladies rares (INTEGRARE), Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Généthon-École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL), Généthon, Chromatin and gene regulation during development (Equipe Inserm U1163), Imagine - Institut des maladies génétiques (IHU) (Imagine - U1163), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Plateforme Bioinformatique [SFR Necker] (BIP-D), Structure Fédérative de Recherche Necker (SFR Necker - UMS 3633 / US24), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université de Paris (UP), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université de Paris (UP), Rice University [Houston], Stanford University, Università degli Studi di Modena e Reggio Emilia (UNIMORE), École pratique des hautes études (EPHE), Université Paris sciences et lettres (PSL)-Université Paris sciences et lettres (PSL)-Université d'Évry-Val-d'Essonne (UEVE)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Généthon, Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité)-Assistance publique - Hôpitaux de Paris (AP-HP) (AP-HP)-Institut National de la Santé et de la Recherche Médicale (INSERM)-Centre National de la Recherche Scientifique (CNRS)-Université Paris Cité (UPCité), Institut National de la Santé et de la Recherche Médicale (INSERM)-Université Paris Cité (UPCité), Università degli Studi di Modena e Reggio Emilia = University of Modena and Reggio Emilia (UNIMORE), CCSD, Accord Elsevier, École Pratique des Hautes Études (EPHE), Chromatin and gene regulation during development, ANR-16-CE18-0004,GETH,L'Édition Génomique comme outil Thérapeutique contre les ß-Hémoglobinopathies(2016)
المصدر: Molecular Therapy
Molecular Therapy, Cell Press, 2019, 27, pp.137-150. ⟨10.1016/j.ymthe.2018.10.008⟩
Molecular Therapy, 2019, 27, pp.137-150. ⟨10.1016/j.ymthe.2018.10.008⟩
بيانات النشر: Elsevier BV, 2019.
سنة النشر: 2019
مصطلحات موضوعية: [SDV]Life Sciences [q-bio], Genetic enhancement, Anemia, Sickle Cell, Biology, Heterogeneous-Nuclear Ribonucleoproteins, Viral vector, 03 medical and health sciences, 0302 clinical medicine, Genome editing, CRISPR-Associated Protein 9, hemic and lymphatic diseases, Drug Discovery, Genetics, Humans, CRISPR, Progenitor cell, Molecular Biology, 030304 developmental biology, Gene Editing, Pharmacology, 0303 health sciences, Cas9, beta-Thalassemia, Genetic Therapy, Hematopoietic Stem Cells, 3. Good health, Cell biology, Hemoglobinopathies, [SDV] Life Sciences [q-bio], Haematopoiesis, 030220 oncology & carcinogenesis, Molecular Medicine, Original Article, CRISPR-Cas Systems, Stem cell, Plasmids, RNA, Guide, Kinetoplastida, CRISPR/Cas9 delivery, genome editing, β-hemoglobinopathies, Drug Discovery3003 Pharmaceutical Science
الوصف: International audience; Editing the beta-globin locus in hematopoietic stem cells is an alternative therapeutic approach for gene therapy of beta-thalassemia and sickle cell disease. Using the CRISPR/Cas9 system, we genetically modified human hematopoietic stem and progenitor cells (HSPCs) to mimic the large rearrangements in the beta-globin locus associated with hereditary persistence of fetal hemoglobin (HPFH), a condition that mitigates the clinical phenotype of patients with beta-hemoglobinopathies. We optimized and compared the efficiency of plasmid-, lentiviral vector (LV)-, RNA-, and ribonucleoprotein complex (RNP)-based methods to deliver the CRISPR/Cas9 system into HSPCs. Plasmid delivery of Cas9 and gRNA pairs targeting two HPFH-like regions led to high frequency of genomic rearrangements and HbF reactivation in erythroblasts derived from sorted, Cas9(+) HSPCs but was associated with significant cell toxicity. RNA-mediated delivery of CRISPR/Cas9 was similarly toxic but much less efficient in editing the beta-globin locus. Transduction of HSPCs by LVs expressing Cas9 and gRNA pairs was robust and minimally toxic but resulted in poor genome-editing efficiency. Ribonucleoprotein (RNP)-based delivery of CRISPR/Cas9 exhibited a good balance between cytotoxicity and efficiency of genomic rearrangements as compared to the other delivery systems and resulted in HbF upregulation in erythroblasts derived from unselected edited HSPCs.
وصف الملف: application/pdf
تدمد: 1525-0016
1525-0024
الوصول الحر: https://explore.openaire.eu/search/publication?articleId=doi_dedup___::5877ed9da54aab838a3dc0587cdf6277Test
https://doi.org/10.1016/j.ymthe.2018.10.008Test
حقوق: OPEN
رقم الانضمام: edsair.doi.dedup.....5877ed9da54aab838a3dc0587cdf6277
قاعدة البيانات: OpenAIRE
الوصف
تدمد:15250016
15250024