Vascular Delivery of rAAVrh74.MCK.GALGT2 to the Gastrocnemius Muscle of the Rhesus Macaque Stimulates the Expression of Dystrophin and Laminin α2 Surrogates
Vascular Delivery of rAAVrh74.MCK.GALGT2 to the Gastrocnemius Muscle of the Rhesus Macaque Stimulates the Expression of Dystrophin and Laminin α2 Surrogates
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DOI：
10.1038/mt.2013.246
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发表时间：
2014-04-01
影响因子：
12.4
通讯作者：
Martin, Paul T.
中科院分区：
文献类型：
--
作者：
Chicoine, Louis G.;Rodino-Klapac, Louise R.;Martin, Paul T.
GALGT 2在骨骼肌中的过表达可以刺激α肌营养不良聚糖的糖基化和正常突触肌营养不良聚糖结合蛋白的上调，其中一些蛋白是肌营养不良蛋白和层粘连蛋白α 2替代物，已知其对几种形式的肌营养不良症具有治疗作用。本文介绍了血管输送GALGT 2基因治疗的大型动物模型，恒河猴。重组腺相关病毒，恒河猴血清型74（rAAVrh 74），被用来提供GALGT 2通过股动脉到腓肠肌使用隔离局灶肢体灌注方法。在具有低预先存在的抗rAAVrh 74血清抗体的猕猴中，治疗后GALGT 2表达平均为44 +/-4%的肌纤维，并且在具有高预先存在的rAAVrh 74免疫力的猕猴中，表达降低至9 +/-4%的肌纤维（P < 0.001;每组n = 12）。这是一个病例，但免疫抑制剂，包括泼尼松龙，他克莫司，和霉酚酸酯的添加。GALGT 2处理的猕猴肌肉显示肌营养不良蛋白聚糖的糖基化增加，肌营养不良蛋白和层粘连蛋白α 2替代蛋白（包括utrophin、plectin 1、agrin和层粘连蛋白α 5）的表达增加。这些实验证明了在血管递送后用rAAVrh74.MCK.GALGT 2成功转导恒河猴肌肉，并诱导了被认为在几种形式的肌营养不良中具有治疗性的分子变化。
Overexpression of GALGT2 in skeletal muscle can stimulate the glycosylation of alpha dystroglycan and the upregulation of normally synaptic dystroglycan-binding proteins, some of which are dystrophin and laminin alpha 2 surrogates known to be therapeutic for several forms of muscular dystrophy. This article describes the vascular delivery of GALGT2 gene therapy in a large animal model, the rhesus macaque. Recombinant adeno-associated virus, rhesus serotype 74 (rAAVrh74), was used to deliver GALGT2 via the femoral artery to the gastrocnemius muscle using an isolated focal limb perfusion method. GALGT2 expression averaged 44 +/- 4% of myofibers after treatment in macaques with low preexisting anti-rAAVrh74 serum antibodies, and expression was reduced to 9 +/- 4% of myofibers in macaques with high preexisting rAAVrh74 immunity (P < 0.001; n = 12 per group). This was the case regard-less of the addition of immunosuppressants, including prednisolone, tacrolimus, and mycophenolate mofetil. GALGT2-treated macaque muscles showed increased glycosylation of a dystroglycan and increased expression of dystrophin and laminin alpha 2 surrogate proteins, including utrophin, plectin1, agrin, and laminin alpha 5. These experiments demonstrate successful transduction of rhesus macaque muscle with rAAVrh74.MCK.GALGT2 after vascular delivery and induction of molecular changes thought to be therapeutic in several forms of muscular dystrophy.