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Genetically blocking HPD via CRISPR-Cas9 protects against lethal liver injury in a pig model of tyrosinemia type I

Hereditary tyrosinemia type I (HT1) results from the loss of fumarylacetoacetate hydrolase (FAH) activity and can lead to lethal liver injury (LLI). Therapeutic options for HT1 remain limited. The FAH(−/−) pig, a well-characterized animal model of HT1, represents a promising candidate for testing no...

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Autores principales: Gu, Peng, Yang, Qin, Chen, Bangzhu, Bie, Ya-nan, Liu, Wen, Tian, Yuguang, Luo, Hongquan, Xu, Tao, Liang, Chunjin, Ye, Xing, Liu, Yan, Tang, Xiangwu, Gu, Weiwang
Formato: Online Artículo Texto
Lenguaje:English
Publicado: American Society of Gene & Cell Therapy 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8099604/
https://www.ncbi.nlm.nih.gov/pubmed/33997102
http://dx.doi.org/10.1016/j.omtm.2021.04.002
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author Gu, Peng
Yang, Qin
Chen, Bangzhu
Bie, Ya-nan
Liu, Wen
Tian, Yuguang
Luo, Hongquan
Xu, Tao
Liang, Chunjin
Ye, Xing
Liu, Yan
Tang, Xiangwu
Gu, Weiwang
author_facet Gu, Peng
Yang, Qin
Chen, Bangzhu
Bie, Ya-nan
Liu, Wen
Tian, Yuguang
Luo, Hongquan
Xu, Tao
Liang, Chunjin
Ye, Xing
Liu, Yan
Tang, Xiangwu
Gu, Weiwang
author_sort Gu, Peng
collection PubMed
description Hereditary tyrosinemia type I (HT1) results from the loss of fumarylacetoacetate hydrolase (FAH) activity and can lead to lethal liver injury (LLI). Therapeutic options for HT1 remain limited. The FAH(−/−) pig, a well-characterized animal model of HT1, represents a promising candidate for testing novel therapeutic approaches to treat this condition. Here, we report an improved single-step method to establish a biallelic (FAH(−/−)) mutant porcine model using CRISPR-Cas9 and cytoplasmic microinjection. We also tested the feasibility of rescuing HT1 pigs through inactivating the 4-hydroxyphenylpyruvic acid dioxygenase (HPD) gene, which functions upstream of the pathogenic pathway, rather than by directly correcting the disease-causing gene as occurs with traditional gene therapy. Direct intracytoplasmic delivery of CRISPR-Cas9 targeting HPD before intrauterine death reprogrammed the tyrosine metabolism pathway and protected pigs against FAH deficiency-induced LLI. Characterization of the F1 generation revealed consistent liver-protective features that were germline transmissible. Furthermore, HPD ablation ameliorated oxidative stress and inflammatory responses and restored the gene profile relating to liver metabolism homeostasis. Collectively, this study not only provided a novel large animal model for exploring the pathogenesis of HT1, but also demonstrated that CRISPR-Cas9-mediated HPD ablation alleviated LLI in HT1 pigs and represents a potential therapeutic option for the treatment of HT1.
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spelling pubmed-80996042021-05-14 Genetically blocking HPD via CRISPR-Cas9 protects against lethal liver injury in a pig model of tyrosinemia type I Gu, Peng Yang, Qin Chen, Bangzhu Bie, Ya-nan Liu, Wen Tian, Yuguang Luo, Hongquan Xu, Tao Liang, Chunjin Ye, Xing Liu, Yan Tang, Xiangwu Gu, Weiwang Mol Ther Methods Clin Dev Original Article Hereditary tyrosinemia type I (HT1) results from the loss of fumarylacetoacetate hydrolase (FAH) activity and can lead to lethal liver injury (LLI). Therapeutic options for HT1 remain limited. The FAH(−/−) pig, a well-characterized animal model of HT1, represents a promising candidate for testing novel therapeutic approaches to treat this condition. Here, we report an improved single-step method to establish a biallelic (FAH(−/−)) mutant porcine model using CRISPR-Cas9 and cytoplasmic microinjection. We also tested the feasibility of rescuing HT1 pigs through inactivating the 4-hydroxyphenylpyruvic acid dioxygenase (HPD) gene, which functions upstream of the pathogenic pathway, rather than by directly correcting the disease-causing gene as occurs with traditional gene therapy. Direct intracytoplasmic delivery of CRISPR-Cas9 targeting HPD before intrauterine death reprogrammed the tyrosine metabolism pathway and protected pigs against FAH deficiency-induced LLI. Characterization of the F1 generation revealed consistent liver-protective features that were germline transmissible. Furthermore, HPD ablation ameliorated oxidative stress and inflammatory responses and restored the gene profile relating to liver metabolism homeostasis. Collectively, this study not only provided a novel large animal model for exploring the pathogenesis of HT1, but also demonstrated that CRISPR-Cas9-mediated HPD ablation alleviated LLI in HT1 pigs and represents a potential therapeutic option for the treatment of HT1. American Society of Gene & Cell Therapy 2021-04-09 /pmc/articles/PMC8099604/ /pubmed/33997102 http://dx.doi.org/10.1016/j.omtm.2021.04.002 Text en © 2021 The Authors https://creativecommons.org/licenses/by-nc-nd/4.0/This is an open access article under the CC BY-NC-ND license (http://creativecommons.org/licenses/by-nc-nd/4.0/).
spellingShingle Original Article
Gu, Peng
Yang, Qin
Chen, Bangzhu
Bie, Ya-nan
Liu, Wen
Tian, Yuguang
Luo, Hongquan
Xu, Tao
Liang, Chunjin
Ye, Xing
Liu, Yan
Tang, Xiangwu
Gu, Weiwang
Genetically blocking HPD via CRISPR-Cas9 protects against lethal liver injury in a pig model of tyrosinemia type I
title Genetically blocking HPD via CRISPR-Cas9 protects against lethal liver injury in a pig model of tyrosinemia type I
title_full Genetically blocking HPD via CRISPR-Cas9 protects against lethal liver injury in a pig model of tyrosinemia type I
title_fullStr Genetically blocking HPD via CRISPR-Cas9 protects against lethal liver injury in a pig model of tyrosinemia type I
title_full_unstemmed Genetically blocking HPD via CRISPR-Cas9 protects against lethal liver injury in a pig model of tyrosinemia type I
title_short Genetically blocking HPD via CRISPR-Cas9 protects against lethal liver injury in a pig model of tyrosinemia type I
title_sort genetically blocking hpd via crispr-cas9 protects against lethal liver injury in a pig model of tyrosinemia type i
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8099604/
https://www.ncbi.nlm.nih.gov/pubmed/33997102
http://dx.doi.org/10.1016/j.omtm.2021.04.002
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