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Deimmunization for gene therapy: host matching of synthetic zinc finger constructs enables long-term mutant Huntingtin repression in mice
dc.contributor.author | Agustín-Pavón, Carmen | |
dc.contributor.author | Mielcarek, Michal | |
dc.contributor.author | Garriga-Canut, Mireia | |
dc.contributor.author | Isalan, Mark | |
dc.date.accessioned | 2016-09-19T09:38:59Z | |
dc.date.available | 2016-09-19T09:38:59Z | |
dc.date.issued | 2016 | |
dc.identifier.citation | AGUSTÍN-PAVÓN, Carmen, et al. Deimmunization for gene therapy: host matching of synthetic zinc finger constructs enables long-term mutant Huntingtin repression in mice. Molecular Neurodegeneration, 2016, vol. 11, no 1, p. 1-16 | ca_CA |
dc.identifier.issn | 1750-1326 | |
dc.identifier.uri | http://hdl.handle.net/10234/162656 | |
dc.description.abstract | Background: Synthetic zinc finger (ZF) proteins can be targeted to desired DNA sequences and are useful tools for gene therapy. We recently developed a ZF transcription repressor (ZF-KOX1) able to bind to expanded DNA CAG-repeats in the huntingtin (HTT) gene, which are found in Huntington’s disease (HD). This ZF acutely repressed mutant HTT expression in a mouse model of HD and delayed neurological symptoms (clasping) for up to 3 weeks. In the present work, we sought to develop a long-term single-injection gene therapy approach in the brain. Method: Since non-self proteins can elicit immune and inflammatory responses, we designed a host-matched analogue of ZF-KOX1 (called mZF-KRAB), to treat mice more safely in combination with rAAV vector delivery. We also tested a neuron-specific enolase promoter (pNSE), which has been reported as enabling long-term transgene expression, to see whether HTT repression could be observed for up to 6 months after AAV injection in the brain. Results: After rAAV vector delivery, we found that non-self proteins induce significant inflammatory responses in the brain, in agreement with previous studies. Specifically, microglial cells were activated at 4 and 6 weeks after treatment with non-host-matched ZF-KOX1 or GFP, respectively, and this was accompanied by a moderate neuronal loss. In contrast, the host-matched mZF-KRAB did not provoke these effects. Nonetheless, we found that using a pCAG promoter (CMV early enhancer element and the chicken β-actin promoter) led to a strong reduction in ZF expression by 6 weeks after injection. We therefore tested a new non-viral promoter to see whether the host-adapted ZF expression could be sustained for a longer time. Vectorising mZF-KRAB with a promoter-enhancer from neuron-specific enolase (Eno2, rat) resulted in up to 77 % repression of mutant HTT in whole brain, 3 weeks after bilateral intraventricular injection of 1010 virions. Importantly, repressions of 48 % and 23 % were still detected after 12 and 24 weeks, respectively, indicating that longer term effects are possible. Conclusion: Host-adapted ZF-AAV constructs displayed a reduced toxicity and a non-viral pNSE promoter improved long-term ZF protein expression and target gene repression. The optimized constructs presented here have potential for treating HD. | ca_CA |
dc.description.sponsorShip | Authors were funded by the European Research Council grants: FP7 ERC 201249 ZINC-HUBS and H2020 - ERC-2014-PoC 641232 - Fingers4Cure. MI is funded by a Wellcome Trust UK New Investigator Award WT102944. | ca_CA |
dc.format.extent | 16 p. | ca_CA |
dc.format.mimetype | application/pdf | ca_CA |
dc.language.iso | eng | ca_CA |
dc.publisher | BioMed Central | ca_CA |
dc.relation.isPartOf | Neurodegeneration, 2016, vol. 11, no 1 | ca_CA |
dc.rights | © 2016 The Author(s). Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated. | ca_CA |
dc.rights | Attribution 4.0 International | * |
dc.rights.uri | http://creativecommons.org/licenses/by-sa/4.0/ | * |
dc.subject | monogenetic disease | ca_CA |
dc.subject | gene therapy | ca_CA |
dc.subject | Huntington’s disease | ca_CA |
dc.subject | neurodegenerative disorder | ca_CA |
dc.subject | immune response | ca_CA |
dc.subject | Synthetic transcription factors | ca_CA |
dc.subject | rAAV | ca_CA |
dc.subject | host optimization | ca_CA |
dc.title | Deimmunization for gene therapy: host matching of synthetic zinc finger constructs enables long-term mutant Huntingtin repression in mice | ca_CA |
dc.type | info:eu-repo/semantics/article | ca_CA |
dc.identifier.doi | http://dx.doi.org/10.1186/s13024-016-0128-x | |
dc.rights.accessRights | info:eu-repo/semantics/openAccess | ca_CA |
dc.relation.publisherVersion | http://molecularneurodegeneration.biomedcentral.com/articles/10.1186/s13024-016-0128-x | ca_CA |
dc.type.version | info:eu-repo/semantics/publishedVersion |
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