Preclinical Systemic Delivery of Adeno-Associated α-Sarcoglycan Gene Transfer for Limb-Girdle Muscular Dystrophy.
Preclinical Systemic Delivery of Adeno-Associated α-Sarcoglycan Gene Transfer for Limb-Girdle Muscular Dystrophy.
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DOI：
10.1089/hum.2019.199
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发表时间：
2021-04
影响因子：
4.2
通讯作者：
Rodino-Klapac LR
中科院分区：
文献类型：
--
作者：
Griffin DA;Pozsgai ER;Heller KN;Potter RA;Peterson EL;Rodino-Klapac LR
肢带型肌营养不良症2D/R3（LGMD 2D/R3）是一种进行性肌营养不良症，表现为肌无力，呼吸异常，在罕见情况下还有心肌病。LGMD 2D/R3是由SGCA基因突变引起的，导致蛋白质丢失和肌营养不良蛋白相关糖蛋白复合物的一些或所有组分的伴随丢失。sgca-null（sgca-/-）小鼠重现了LGMD 2D/R3患者的临床表型，包括营养不良特征，如肌肉坏死和纤维化、血清肌酸激酶（CK）升高以及绝对肌力和运动活性的产生减少。因此，sgca−/−小鼠为测试基因转移的安全性和有效性提供了相关模型。我们设计了一种自身互补的AAVrh 74载体，其含有由肌肉特异性启动子缩短的肌肉肌酸激酶（tMCK）驱动的密码子优化的全长人SGCA（hSGCA）转基因。在本报告中，我们使用剂量递增设计在sgca−/−小鼠中检测了的有效性和安全性，以评价与溶剂处理和野生型小鼠相比，单次全身注射1.0 × 1012、3.0 × 1012和6.0 × 1012 vg总剂量的有效性和安全性。在sgca−/−小鼠中，在所有测试剂量下，用处理均导致骨骼肌肌膜上α-肌聚糖蛋白的稳健表达。此外，还能有效改善sgca−/−小鼠肢体和膈肌的组织病理学，如纤维化减少、中央成核和肌纤维大小正常化所示。这些分子变化伴随着膈肌和胫骨前肌的比力产生显著增加，防止离心力损失，以及血清CK降低。与溶剂处理的sgca−/−小鼠相比，所有剂量的载体处理的小鼠的运动活性均得到改善。最后，在血清化学检查和大体尸检中未检测到载体毒性。总的来说，这些发现为在临床环境中全身递送用于治疗LGMD 2D/R3提供了支持。
Limb-girdle muscular dystrophy type 2D/R3 (LGMD2D/R3) is a progressive muscular dystrophy that manifests with muscle weakness, respiratory abnormalities, and in rare cases cardiomyopathy. LGMD2D/R3 is caused by mutations in the SGCA gene resulting in loss of protein and concomitant loss of some or all components of the dystrophin-associated glycoprotein complex. The sgca-null (sgca−/−) mouse recapitulates the clinical phenotype of patients with LGMD2D/R3, including dystrophic features such as muscle necrosis and fibrosis, elevated serum creatine kinase (CK), and reduction in the generation of absolute muscle force and locomotor activity. Thus, sgca−/− mice provide a relevant model to test the safety and efficacy of gene transfer. We designed a self-complementary AAVrh74 vector containing a codon-optimized full-length human SGCA (hSGCA) transgene driven by a muscle-specific promoter, shortened muscle creatine kinase (tMCK). In this report, we test the efficacy and safety of in sgca−/− mice using a dose-escalation design to evaluate a single systemic injection of 1.0 × 1012, 3.0 × 1012, and 6.0 × 1012 vg total dose compared with vehicle-treatment and wild-type mice. In sgca−/− mice, treatment with resulted in robust expression of α-sarcoglycan protein at the sarcolemma membrane in skeletal muscle at all doses tested. In addition, was effective in improving the histopathology of limb and diaphragm muscle of sgca−/− mice, as indicated by reductions in fibrosis, central nucleation, and normalization of myofiber size. These molecular changes were concomitant with significant increases in specific force generation in the diaphragm and tibialis anterior muscle, protection against eccentric force loss, and reduction in serum CK. Locomotor activity was improved at all doses of vector-treated compared with vehicle-treated sgca−/− mice. Lastly, vector toxicity was not detected in a serum chemistry panel and by gross necropsy. Collectively, these findings provide support for a systemic delivery of in a clinical setting for the treatment of LGMD2D/R3.
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肌肉营养不良的动物模型显示出不同的肌膜破坏模式。
影响因子：
7.8
作者：
Straub, V;Rafael, JA;Chamberlain, JS;Campbell, KP
通讯作者：
Campbell, KP
影响因子：
7.4
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Mendell, Jerry R.
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影响因子：
5.1
作者：
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通讯作者：
Samulski, RJ
影响因子：
4.2
作者：
Potter RA;Griffin DA;Sondergaard PC;Johnson RW;Pozsgai ER;Heller KN;Peterson EL;Lehtimäki KK;Windish HP;Mittal PJ;Albrecht DE;Mendell JR;Rodino-Klapac LR
通讯作者：
Rodino-Klapac LR