Adeno-associated viral-mediated LARGE gene therapy rescues the muscular dystrophic phenotype in mouse models of dystroglycanopathy.
Adeno-associated viral-mediated LARGE gene therapy rescues the muscular dystrophic phenotype in mouse models of dystroglycanopathy.
复制标题
腺相关病毒介导的大基因疗法可挽救肌营养不良症小鼠模型中的肌营养不良表型。
DOI：
10.1089/hum.2012.084
复制
发表时间：
2013
影响因子：
4.2
通讯作者：
Hu,Huaiyu
中科院分区：
文献类型：
--
作者：
Yu,Miao;He,Yonglin;Wang,Kejian;Zhang,Peng;Zhang,Shengle;Hu,Huaiyu
糖营养不良症是一组先天性肌营养不良症（CMD），通常由编码糖基转移酶的基因突变引起，导致α-糖营养不良（α-DG）的低糖基化并降低其细胞外基质结合活性。过表达LARGE（以前称为样糖基转移酶）在细胞中产生细胞外基质结合碳水化合物表位，这些细胞不仅在LARGE而且在其他糖基转移酶（包括POMT1， POMGnT1和fukutin）中发生cmd引起的突变，从而创造了一种“一刀切”的基因治疗的可能性。为了确定LARGE基因治疗的可行性，将血清型9型腺相关病毒载体（AAV9-LARGE）注射到两种CMD小鼠模型的新生儿心脏内：天然LARGE突变体largemyd小鼠和蛋白o-甘露糖n -乙酰氨基葡萄糖转移酶1 （POMGnT1）敲除小鼠。AAV9-LARGE病毒处理可部分恢复α-DG糖基化和配体结合活性。骨骼肌肌营养不良表型得到改善，纤维化、坏死和中央核数量显著减少，运动功能改善。这些结果表明，aav9介导的LARGE过表达体内基因治疗可以有效地恢复α-DG的功能性糖基化，并在LARGE和POMGnT1缺乏的情况下挽救肌营养不良表型，这证明了体内LARGE基因治疗可能广泛适用于糖营养不良症。
Dystroglycanopathies are a group of congenital muscular dystrophies (CMD) often caused by mutations in genes encoding glycosyltransferases that lead to hypoglycosylation of α-dystroglycan (α-DG) and reduce its extracellular matrix-binding activity. OverexpressingLARGE(formerly known as like-glycosyltransferase) generates an extracellular matrix-binding carbohydrate epitope in cells with CMD-causing mutations in not only LARGE but also other glycosyltransferases, including POMT1, POMGnT1, and fukutin, creating the possibilities of a one-for-all gene therapy. To determine the feasibility of LARGE gene therapy, a serotype 9 adeno-associated viral vector for overexpressing LARGE (AAV9-LARGE) was injected intracardially into newborns of two mouse models of CMD: the natural LARGE mutant Largemydmice and protein O-mannose N-acetylglucosaminyltransferase 1 (POMGnT1) knockout mice. AAV9-LARGE virus treatment yielded partial restoration of α-DG glycosylation and ligand-binding activity. The muscular dystrophy phenotype in skeletal muscles was ameliorated as revealed by significantly reduced fibrosis, necrosis, and numbers of centrally located nuclei with improved motor function. These results indicate that LARGE overexpressionin vivoby AAV9-mediated gene therapy is effective at restoring functional glycosylation of α-DG and rescuing the muscular dystrophy phenotype in deficiency of not only LARGE but also POMGnT1, providing evidence thatin vivoLARGEgene therapy may be broadly useful in dystroglycanopathies.