|
OKANO Teruo
Department Research Institutes and Facilities, Research Institutes and Facilities Position |
|
| Article types | Original article |
| Language | English |
| Peer review | Peer reviewed |
| Title | Development of alternative gene transfer techniques for ex vivo and in vivo gene therapy in a canine model. |
| Journal | Formal name：Regenerative Therapy ISSN code：2352-3204 |
| Domestic / Foregin | Foregin |
| Volume, Issue, Page | 18,pp.347-354 |
| Author and coauthor | NODA Masashi†, TATSUMI Kohei*, MATSUI Hideto, MATSUNARI Yasunori, SATO Takeshi, FUKUOKA Yasushi, HOTTA Akitsu, OKANO Teruo, KICHIKAWA Kimihiko, SUGIMOTO Mitsuhiko, SHIMA Midori, NISHIO Kenji |
| Publication date | 2021/12 |
| Summary | Introduction:Gene therapy have recently attracted much attention as a curative therapeutic option for inherited single gene disorders such as hemophilia. Hemophilia is a hereditary bleeding disorder caused by the deficiency of clotting activity of factor VIII (FVIII) or factor IX (FIX), and gene therapy for hemophilia using viral vector have been vigorously investigated worldwide. Toward further advancement of gene therapy for hemophilia, we have previously developed and validated the efficacy of novel two types of gene transfer technologies using a mouse model of hemophilia A. Here we investigated the efficacy and safety of the technologies in canine model. Especially, validations of technical procedures of the gene transfers for dogs were focused.Methods:Green fluorescence protein (GFP) gene were transduced into normal beagle dogs by ex vivo and in vivo gene transfer techniques. For ex vivo gene transfer, blood outgrowth endothelial cells (BOECs) derived from peripheral blood of normal dogs were transduced with GFP gene using lentivirus vector, propagated, fabricated as cell sheets, then implanted onto the omentum of the same dogs. For in vivo gene transfer, normal dogs were subjected to GFP gene transduction with non-viral piggyBac vector by liver-targeted hydrodynamic injections.Results:No major adverse events were observed during the gene transfers in both gene transfer systems. As for ex vivo gene transfer, histological findings from the omental biopsy performed 4 weeks after implantation revealed the tube formation by implanted GFP-positive BOECs in the sub-adipose tissue layer without any inflammatory findings, and the detected GFP signals were maintained over 6 months. Regarding in vivo gene transfer, analyses of liver biopsy samples revealed more than 90% of liver cells were positive for GFP signals in the injected liver lobes 1 week after gene transfers, then the signals gradually declined overtime.Conclusions:Two types of gene transfer techniques were su |
| DOI | 10.1016/j.reth.2021.08.009 |
| PMID | 34584911 |