单一 rAAV3B-AAT 载体在犬科猕猴中沉默和替代α-1 抗胰蛋白酶的生物分布和安全性

IF 4.6 2区 医学 Q2 MEDICINE, RESEARCH & EXPERIMENTAL
Meghan Blackwood, Alisha M. Gruntman, Qiushi Tang, Debora Pires-Ferreira, Darcy Reil, Oleksandr Kondratov, Damien Marsic, Sergei Zolotukin, Gwladys Gernoux, Allison M. Keeler, Christian Mueller, Terence R. Flotte
{"title":"单一 rAAV3B-AAT 载体在犬科猕猴中沉默和替代α-1 抗胰蛋白酶的生物分布和安全性","authors":"Meghan Blackwood, Alisha M. Gruntman, Qiushi Tang, Debora Pires-Ferreira, Darcy Reil, Oleksandr Kondratov, Damien Marsic, Sergei Zolotukin, Gwladys Gernoux, Allison M. Keeler, Christian Mueller, Terence R. Flotte","doi":"10.1016/j.omtm.2024.101200","DOIUrl":null,"url":null,"abstract":"<p>Alpha-1 antitrypsin deficiency (AATD) is characterized by both chronic lung disease due to loss of wild-type AAT (M-AAT) antiprotease function and liver disease due to toxicity from delayed secretion, polymerization, and aggregation of misfolded mutant AAT (Z-AAT). The ideal gene therapy for AATD should therefore comprise both endogenous Z-AAT suppression and M-AAT overexpression. We designed a dual-function rAAV3B (df-rAAV3B) construct, which was effective at transducing hepatocytes, resulting in a considerable decrease of Z-AAT levels and safe M-AAT augmentation in mice. We optimized df-rAAV3B and created two variants, AAV3B-E12 and AAV3B-G3, to simultaneously enhance the concentration of M-AAT in the bloodstream to therapeutic levels and silence endogenous AAT liver expression in cynomolgus monkey. Our results demonstrate that AAV3b-WT, AAV3B-E12, and AAV3B-G3 were able to transduce the monkey livers and achieve high M-AAT serum levels efficiently and safely. In this non-deficient model, we did not find down-regulation of endogenous AAT. However, the dual-function vector did serve as a potentially “liver-sparing” alternative for high dose liver-mediated AAT gene replacement in the context of underlying liver disease.</p>","PeriodicalId":54333,"journal":{"name":"Molecular Therapy-Methods & Clinical Development","volume":null,"pages":null},"PeriodicalIF":4.6000,"publicationDate":"2024-01-30","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"Biodistribution and safety of a single rAAV3B-AAT vector for silencing and replacement of alpha-1 antitrypsin in Cynomolgus macaques\",\"authors\":\"Meghan Blackwood, Alisha M. Gruntman, Qiushi Tang, Debora Pires-Ferreira, Darcy Reil, Oleksandr Kondratov, Damien Marsic, Sergei Zolotukin, Gwladys Gernoux, Allison M. Keeler, Christian Mueller, Terence R. Flotte\",\"doi\":\"10.1016/j.omtm.2024.101200\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p>Alpha-1 antitrypsin deficiency (AATD) is characterized by both chronic lung disease due to loss of wild-type AAT (M-AAT) antiprotease function and liver disease due to toxicity from delayed secretion, polymerization, and aggregation of misfolded mutant AAT (Z-AAT). The ideal gene therapy for AATD should therefore comprise both endogenous Z-AAT suppression and M-AAT overexpression. We designed a dual-function rAAV3B (df-rAAV3B) construct, which was effective at transducing hepatocytes, resulting in a considerable decrease of Z-AAT levels and safe M-AAT augmentation in mice. We optimized df-rAAV3B and created two variants, AAV3B-E12 and AAV3B-G3, to simultaneously enhance the concentration of M-AAT in the bloodstream to therapeutic levels and silence endogenous AAT liver expression in cynomolgus monkey. Our results demonstrate that AAV3b-WT, AAV3B-E12, and AAV3B-G3 were able to transduce the monkey livers and achieve high M-AAT serum levels efficiently and safely. In this non-deficient model, we did not find down-regulation of endogenous AAT. However, the dual-function vector did serve as a potentially “liver-sparing” alternative for high dose liver-mediated AAT gene replacement in the context of underlying liver disease.</p>\",\"PeriodicalId\":54333,\"journal\":{\"name\":\"Molecular Therapy-Methods & Clinical Development\",\"volume\":null,\"pages\":null},\"PeriodicalIF\":4.6000,\"publicationDate\":\"2024-01-30\",\"publicationTypes\":\"Journal Article\",\"fieldsOfStudy\":null,\"isOpenAccess\":false,\"openAccessPdf\":\"\",\"citationCount\":\"0\",\"resultStr\":null,\"platform\":\"Semanticscholar\",\"paperid\":null,\"PeriodicalName\":\"Molecular Therapy-Methods & Clinical Development\",\"FirstCategoryId\":\"3\",\"ListUrlMain\":\"https://doi.org/10.1016/j.omtm.2024.101200\",\"RegionNum\":2,\"RegionCategory\":\"医学\",\"ArticlePicture\":[],\"TitleCN\":null,\"AbstractTextCN\":null,\"PMCID\":null,\"EPubDate\":\"\",\"PubModel\":\"\",\"JCR\":\"Q2\",\"JCRName\":\"MEDICINE, RESEARCH & EXPERIMENTAL\",\"Score\":null,\"Total\":0}","platform":"Semanticscholar","paperid":null,"PeriodicalName":"Molecular Therapy-Methods & Clinical Development","FirstCategoryId":"3","ListUrlMain":"https://doi.org/10.1016/j.omtm.2024.101200","RegionNum":2,"RegionCategory":"医学","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"MEDICINE, RESEARCH & EXPERIMENTAL","Score":null,"Total":0}
引用次数: 0

摘要

α-1抗胰蛋白酶缺乏症(AATD)的特点是,野生型AAT(M-AAT)抗蛋白酶功能丧失导致慢性肺部疾病,而错误折叠的突变型AAT(Z-AAT)的延迟分泌、聚合和聚集则导致肝脏疾病。因此,AATD 的理想基因疗法应包括内源性 Z-AAT 抑制和 M-AAT 过度表达。我们设计了一种双重功能的 rAAV3B(df-rAAV3B)构建体,它能有效转导肝细胞,使小鼠体内的 Z-AAT 水平显著下降,M-AAT 安全增强。我们对 df-rAAV3B 进行了优化,并创建了两个变体:AAV3B-E12 和 AAV3B-G3,以同时将血液中的 M-AAT 浓度提高到治疗水平,并抑制犬科猴肝脏中内源性 AAT 的表达。我们的研究结果表明,AAV3b-WT、AAV3B-E12 和 AAV3B-G3 能够转导猴肝,并高效、安全地达到较高的 M-AAT 血清水平。在这种非缺陷模型中,我们没有发现内源性 AAT 的下调。不过,在有潜在肝病的情况下,双功能载体确实是一种潜在的 "保肝 "替代品,可用于大剂量肝脏介导的 AAT 基因替代。
本文章由计算机程序翻译,如有差异,请以英文原文为准。

Biodistribution and safety of a single rAAV3B-AAT vector for silencing and replacement of alpha-1 antitrypsin in Cynomolgus macaques

Biodistribution and safety of a single rAAV3B-AAT vector for silencing and replacement of alpha-1 antitrypsin in Cynomolgus macaques

Alpha-1 antitrypsin deficiency (AATD) is characterized by both chronic lung disease due to loss of wild-type AAT (M-AAT) antiprotease function and liver disease due to toxicity from delayed secretion, polymerization, and aggregation of misfolded mutant AAT (Z-AAT). The ideal gene therapy for AATD should therefore comprise both endogenous Z-AAT suppression and M-AAT overexpression. We designed a dual-function rAAV3B (df-rAAV3B) construct, which was effective at transducing hepatocytes, resulting in a considerable decrease of Z-AAT levels and safe M-AAT augmentation in mice. We optimized df-rAAV3B and created two variants, AAV3B-E12 and AAV3B-G3, to simultaneously enhance the concentration of M-AAT in the bloodstream to therapeutic levels and silence endogenous AAT liver expression in cynomolgus monkey. Our results demonstrate that AAV3b-WT, AAV3B-E12, and AAV3B-G3 were able to transduce the monkey livers and achieve high M-AAT serum levels efficiently and safely. In this non-deficient model, we did not find down-regulation of endogenous AAT. However, the dual-function vector did serve as a potentially “liver-sparing” alternative for high dose liver-mediated AAT gene replacement in the context of underlying liver disease.

求助全文
通过发布文献求助,成功后即可免费获取论文全文。 去求助
来源期刊
Molecular Therapy-Methods & Clinical Development
Molecular Therapy-Methods & Clinical Development Biochemistry, Genetics and Molecular Biology-Molecular Biology
CiteScore
9.90
自引率
4.30%
发文量
163
审稿时长
12 weeks
期刊介绍: The aim of Molecular Therapy—Methods & Clinical Development is to build upon the success of Molecular Therapy in publishing important peer-reviewed methods and procedures, as well as translational advances in the broad array of fields under the molecular therapy umbrella. Topics of particular interest within the journal''s scope include: Gene vector engineering and production, Methods for targeted genome editing and engineering, Methods and technology development for cell reprogramming and directed differentiation of pluripotent cells, Methods for gene and cell vector delivery, Development of biomaterials and nanoparticles for applications in gene and cell therapy and regenerative medicine, Analysis of gene and cell vector biodistribution and tracking, Pharmacology/toxicology studies of new and next-generation vectors, Methods for cell isolation, engineering, culture, expansion, and transplantation, Cell processing, storage, and banking for therapeutic application, Preclinical and QC/QA assay development, Translational and clinical scale-up and Good Manufacturing procedures and process development, Clinical protocol development, Computational and bioinformatic methods for analysis, modeling, or visualization of biological data, Negotiating the regulatory approval process and obtaining such approval for clinical trials.
×
引用
GB/T 7714-2015
复制
MLA
复制
APA
复制
导出至
BibTeX EndNote RefMan NoteFirst NoteExpress
×
提示
您的信息不完整,为了账户安全,请先补充。
现在去补充
×
提示
您因"违规操作"
具体请查看互助需知
我知道了
×
提示
确定
请完成安全验证×
copy
已复制链接
快去分享给好友吧!
我知道了
右上角分享
点击右上角分享
0
联系我们:info@booksci.cn Book学术提供免费学术资源搜索服务,方便国内外学者检索中英文文献。致力于提供最便捷和优质的服务体验。 Copyright © 2023 布克学术 All rights reserved.
京ICP备2023020795号-1
ghs 京公网安备 11010802042870号
Book学术文献互助
Book学术文献互助群
群 号:481959085
Book学术官方微信