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.
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单个 rAAV3B-AAT 载体在食蟹猴中沉默和替代 α-1 抗胰蛋白酶的生物分布和安全性。
DOI：
10.1016/j.omtm.2024.101200
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发表时间：
2024
期刊：
影响因子：
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通讯作者：
Flotte,TerenceR
中科院分区：
文献类型：
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作者：
Blackwood,Meghan;Gruntman,AlishaM;Tang,Qiushi;Pires-Ferreira,Debora;Reil,Darcy;Kondratov,Oleksandr;Marsic,Damien;Zolotukhin,Sergei;Gernoux,Gwladys;Keeler,AllisonM;Mueller,Christian;Flotte,TerenceR
α -1抗胰蛋白酶缺缺症（AATD）的特征是由于野生型AAT （M-AAT）抗蛋白酶功能丧失引起的慢性肺部疾病，以及由于错误折叠突变型AAT （Z-AAT）的分泌、聚合和聚集延迟引起的毒性导致的肝脏疾病。因此，理想的AATD基因治疗应该包括内源性Z-AAT抑制和M-AAT过表达。我们设计了一种双功能的rAAV3B （df-rAAV3B）构建体，它能有效地转导肝细胞，导致小鼠体内Z-AAT水平显著降低，M-AAT安全增加。我们对df-rAAV3B进行了优化，并创建了AAV3B-E12和AAV3B-G3两个变体，同时将食食猴血液中M-AAT的浓度提高到治疗水平，并抑制内源性AAT在肝脏中的表达。结果表明，AAV3b-WT、AAV3B-E12和AAV3B-G3能够高效、安全地转导猴子肝脏并获得高水平的血清M-AAT。在这个非缺陷模型中，我们没有发现内源性AAT下调。然而，在潜在肝病的情况下，双功能载体确实可以作为大剂量肝脏介导的AAT基因替代的潜在“保肝”选择。
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 monkeys. 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 nondeficient model, we did not find downregulation 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.