Cargando…

Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response

AAV-delivered CRISPR/Cas9 (AAV-CRISPR) has shown promising potentials in preclinical models to efficiently insert therapeutic gene sequences in somatic tissues. However, the AAV input doses required were prohibitively high and posed serious risk of toxicity. Here, we performed AAV-CRISPR mediated ho...

Descripción completa

Detalles Bibliográficos
Autores principales: He, Xiangjun, Zhang, Zhenjie, Xue, Junyi, Wang, Yaofeng, Zhang, Siqi, Wei, Junkang, Zhang, Chenzi, Wang, Jue, Urip, Brian Anugerah, Ngan, Chun Christopher, Sun, Junjiang, Li, Yuefeng, Lu, Zhiqian, Zhao, Hui, Pei, Duanqing, Li, Chi-Kong, Feng, Bo
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700734/
https://www.ncbi.nlm.nih.gov/pubmed/36434000
http://dx.doi.org/10.1038/s41467-022-34898-y
_version_ 1784839377844699136
author He, Xiangjun
Zhang, Zhenjie
Xue, Junyi
Wang, Yaofeng
Zhang, Siqi
Wei, Junkang
Zhang, Chenzi
Wang, Jue
Urip, Brian Anugerah
Ngan, Chun Christopher
Sun, Junjiang
Li, Yuefeng
Lu, Zhiqian
Zhao, Hui
Pei, Duanqing
Li, Chi-Kong
Feng, Bo
author_facet He, Xiangjun
Zhang, Zhenjie
Xue, Junyi
Wang, Yaofeng
Zhang, Siqi
Wei, Junkang
Zhang, Chenzi
Wang, Jue
Urip, Brian Anugerah
Ngan, Chun Christopher
Sun, Junjiang
Li, Yuefeng
Lu, Zhiqian
Zhao, Hui
Pei, Duanqing
Li, Chi-Kong
Feng, Bo
author_sort He, Xiangjun
collection PubMed
description AAV-delivered CRISPR/Cas9 (AAV-CRISPR) has shown promising potentials in preclinical models to efficiently insert therapeutic gene sequences in somatic tissues. However, the AAV input doses required were prohibitively high and posed serious risk of toxicity. Here, we performed AAV-CRISPR mediated homology-independent knock-in at a new target site in mAlb 3’UTR and demonstrated that single dose of AAVs enabled long-term integration and expression of hF9 transgene in both adult and neonatal hemophilia B mice (mF9 −/−), yielding high levels of circulating human Factor IX (hFIX) and stable hemostasis restoration during entire 48-week observation period. Furthermore, we achieved hemostasis correction with a significantly lower AAV dose (2 × 10(9) vg/neonate and 1 × 10(10) vg/adult mouse) through liver-specific gene knock-in using hyperactive hF9(R338L) variant. The plasma antibodies against Cas9 and AAV in the neonatal mice receiving low-dose AAV-CRISPR were negligible, which lent support to the development of AAV-CRISPR mediated somatic knock-in for treating inherited diseases.
format Online
Article
Text
id pubmed-9700734
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher Nature Publishing Group UK
record_format MEDLINE/PubMed
spelling pubmed-97007342022-11-27 Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response He, Xiangjun Zhang, Zhenjie Xue, Junyi Wang, Yaofeng Zhang, Siqi Wei, Junkang Zhang, Chenzi Wang, Jue Urip, Brian Anugerah Ngan, Chun Christopher Sun, Junjiang Li, Yuefeng Lu, Zhiqian Zhao, Hui Pei, Duanqing Li, Chi-Kong Feng, Bo Nat Commun Article AAV-delivered CRISPR/Cas9 (AAV-CRISPR) has shown promising potentials in preclinical models to efficiently insert therapeutic gene sequences in somatic tissues. However, the AAV input doses required were prohibitively high and posed serious risk of toxicity. Here, we performed AAV-CRISPR mediated homology-independent knock-in at a new target site in mAlb 3’UTR and demonstrated that single dose of AAVs enabled long-term integration and expression of hF9 transgene in both adult and neonatal hemophilia B mice (mF9 −/−), yielding high levels of circulating human Factor IX (hFIX) and stable hemostasis restoration during entire 48-week observation period. Furthermore, we achieved hemostasis correction with a significantly lower AAV dose (2 × 10(9) vg/neonate and 1 × 10(10) vg/adult mouse) through liver-specific gene knock-in using hyperactive hF9(R338L) variant. The plasma antibodies against Cas9 and AAV in the neonatal mice receiving low-dose AAV-CRISPR were negligible, which lent support to the development of AAV-CRISPR mediated somatic knock-in for treating inherited diseases. Nature Publishing Group UK 2022-11-25 /pmc/articles/PMC9700734/ /pubmed/36434000 http://dx.doi.org/10.1038/s41467-022-34898-y Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as 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 images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
He, Xiangjun
Zhang, Zhenjie
Xue, Junyi
Wang, Yaofeng
Zhang, Siqi
Wei, Junkang
Zhang, Chenzi
Wang, Jue
Urip, Brian Anugerah
Ngan, Chun Christopher
Sun, Junjiang
Li, Yuefeng
Lu, Zhiqian
Zhao, Hui
Pei, Duanqing
Li, Chi-Kong
Feng, Bo
Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response
title Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response
title_full Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response
title_fullStr Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response
title_full_unstemmed Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response
title_short Low-dose AAV-CRISPR-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia B mice with subtle antibody response
title_sort low-dose aav-crispr-mediated liver-specific knock-in restored hemostasis in neonatal hemophilia b mice with subtle antibody response
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9700734/
https://www.ncbi.nlm.nih.gov/pubmed/36434000
http://dx.doi.org/10.1038/s41467-022-34898-y
work_keys_str_mv AT hexiangjun lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT zhangzhenjie lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT xuejunyi lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT wangyaofeng lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT zhangsiqi lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT weijunkang lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT zhangchenzi lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT wangjue lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT uripbriananugerah lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT nganchunchristopher lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT sunjunjiang lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT liyuefeng lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT luzhiqian lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT zhaohui lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT peiduanqing lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT lichikong lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse
AT fengbo lowdoseaavcrisprmediatedliverspecificknockinrestoredhemostasisinneonatalhemophiliabmicewithsubtleantibodyresponse