An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque.
An Isolated Limb Infusion Method Allows for Broad Distribution of rAAVrh74.MCK.GALGT2 to Leg Skeletal Muscles in the Rhesus Macaque.
复制标题
离体肢体输注方法允许 rAAVrh74.MCK.GALGT2 广泛分布到恒河猴的腿部骨骼肌。
DOI：
10.1016/j.omtm.2018.06.002
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发表时间：
2018
期刊：
影响因子：
--
通讯作者：
Rodi
中科院分区：
文献类型：
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作者：
Xu,Rui;Jia,Ying;Zygmunt,DeborahA;Cramer,MeganL;Crowe,KellyE;Shao,Guohong;Maki,AgathaE;Guggenheim,HaleyN;Hood,BenjaminC;Griffin,DanielleA;Peterson,Ellyn;Bolon,Brad;Cheatham,JohnP;Cheatham,SharonL;Flanigan,KevinM;Rodi
重组腺相关病毒（rAAV）rh74.MCKgalg2是一种肌肉特异性基因疗法，正在开发用于治疗各种形式的肌肉萎缩症。在这里，我们报告了一种非人类灵长类动物模型的分离肢体输注技术，其中后肢血流使用球囊导管短暂分离，将载体集中在目标腿部肌肉中。2.5 × 1013个载体基因组(vg)/kg/肢的双侧剂量足以在所有腿部肌肉中诱导galgt2诱导的10%-60%的骨骼肌纤维糖基化。与双侧剂量为6 × 1012vg/kg/肢相比，双侧剂量为2.5 × 1013vg/kg/肢时，每微克基因组DNA的载体基因组平均肢宽增加19倍±6倍。单侧剂量为6 × 1013vg/kg/肢时，治疗肢体肌肉比对侧未治疗肢体肌肉增加12±3倍，而对侧未治疗肢体只有在释放到体循环后才接受载体。同一肌肉不同节段之间AAV生物分布的变异性在任何给定剂量下为125%±18%，而在任何给定治疗剂量下同一肌肉之间的变异性为45%±7%。这些实验表明，rAAVrh74.MCK治疗整个腿部肌肉。galg2可以通过肢体隔离输注技术安全地完成，气囊导管可暂时隔离肢体血管，但肌肉内和肌肉间的转导可变性是一个重要问题。
Recombinant adeno-associated virus (rAAV)rh74.MCK.GALGT2is a muscle-specific gene therapy that is being developed to treat forms of muscular dystrophy. Here we report on an isolated limb infusion technique in a non-human primate model, where hindlimb blood flow is transiently isolated using balloon catheters to concentrate vector in targeted leg muscles. A bilateral dose of 2.5 × 1013vector genomes (vg)/kg/limb was sufficient to induce GALGT2-induced glycosylation in 10%–60% of skeletal myofibers in all leg muscles examined. There was a 19-fold ± 6-fold average limb-wide increase in vector genomes per microgram genomic DNA at a bilateral dose of 2.5 × 1013vg/kg/limb compared with a bilateral dose of 6 × 1012vg/kg/limb. A unilateral dose of 6 × 1013vg/kg/limb showed a 12- ± 3-fold increase in treated limb muscles compared to contralateral untreated limb muscles, which received vector only after release into the systemic circulation from the treated limb. Variability in AAV biodistribution between different segments of the same muscle was 125% ± 18% for any given dose, while variability between the same muscle for any given treatment dose was 45% ± 7%. These experiments demonstrate that treatment of muscles throughout the leg with rAAVrh74.MCK.GALGT2can be accomplished safely using an isolated limb infusion technique, where balloon catheters transiently isolate the limb vasculature, but that intra- and inter-muscle transduction variability is a significant issue.