Intrathymic AAV delivery results in therapeutic site-specific integration at TCR loci in mice
In: ISSN: 0006-4971, 2023
Online
academicJournal
Zugriff:
International audience ; Intrathymic delivery of AAV results in vector integration within TCR genes at RAG-induced DNA breaks produced during V(D)J recombination. • This "targeting" approach opens therapeutic avenues for long-term AAV gene transfer in dividing cells without any toxic conditioning. Adeno-associated virus (AAV) vectors have been successfully exploited in gene therapy applications for the treatment of several genetic disorders. AAV is considered an episomal vector, but it has been shown to integrate within the host cell genome after the generation of double-strand DNA breaks or nicks. Although AAV integration raises some safety concerns, it can also provide therapeutic benefit; the direct intrathymic injection of an AAV harboring a therapeutic transgene results in integration in T-cell progenitors and long-term T-cell immunity. To assess the mechanisms of AAV integration, we retrieved and analyzed hundreds of AAV integration sites from lymph node-derived mature T cells and compared these with liver and brain tissue from treated mice. Notably, we found that although AAV integrations in the liver and brain were distributed across the entire mouse genome, >90% of the integrations in T cells were clustered within the T-cell receptor α, β, and γ genes. More precisely, the insertion mapped to DNA breaks created by the enzymatic activity of recombination activating genes (RAGs) during variable, diversity, and joining recombination. Our data indicate that RAG activity during T-cell receptor maturation induces a site-specific integration of AAV genomes and opens new therapeutic avenues for achieving long-term AAV-mediated gene transfer in dividing cells.
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Intrathymic AAV delivery results in therapeutic site-specific integration at TCR loci in mice
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Autor/in / Beteiligte Person: | Calabria, Andrea ; Cipriani, Carlo ; Spinozzi, Giulio ; Rudilosso, Laura ; Esposito, Simona ; Benedicenti, Fabrizio ; Albertini, Alessandra ; Pouzolles, Marie ; Luoni, Mirko ; Giannelli, Serena ; Broccoli, Vania ; Guilbaud, Mickael ; Adjali, Oumeya ; Taylor, Naomi ; Zimmermann, Valérie ; Montini, Eugenio ; Cesana, Daniela ; IRCCS San Raffaele Scientific Institute Milan, Italie ; Institut de Génétique Moléculaire de Montpellier (IGMM) ; Centre National de la Recherche Scientifique (CNRS)-Université de Montpellier (UM) ; San Raffaele Scientific Institute ; Vita-Salute San Raffaele University and Center for Translational Genomics and Bioinformatics ; National Research Council of Italy (CNR) ; Institute of Biology, Molecular Medicine and Nanobiotechnology. Sapienza University of Rome (IBMN) ; Laboratoire de Thérapie Génique Translationnelle des Maladies Génétiques / Translational Research in Gene Therapy - UMR_S 1089 (TARGET) ; Institut National de la Santé et de la Recherche Médicale (INSERM)-Nantes Université - UFR de Médecine et des Techniques Médicales (Nantes Univ - UFR MEDECINE) ; Nantes Université - pôle Santé ; Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ)-Nantes Université - pôle Santé ; Nantes Université (Nantes Univ)-Nantes Université (Nantes Univ) ; National Institutes of Health Bethesda, MD, USA (NIH) |
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Zeitschrift: | ISSN: 0006-4971, 2023 |
Veröffentlichung: | HAL CCSD ; American Society of Hematology, 2023 |
Medientyp: | academicJournal |
DOI: | 10.1182/blood.2022017378 |
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