Elmallah Mai K, Falk Darin J, Nayak Sushrusha, Federico Roland A, Sandhu Milapjit S, Poirier Amy, Byrne Barry J, Fuller David D
Department of Pediatrics, Division of Pulmonary Medicine, College of Medicine, University of Florida, Gainesville, Florida, USA.
1] Department of Pediatrics, Divisions of Cellular and Molecular Therapy and Pediatric Cardiology, College of Medicine, University of Florida, Gainesville, Florida, USA [2] Powell Gene Therapy Center, University of Florida, Gainesville, Florida, USA.
Mol Ther. 2014 Apr;22(4):702-12. doi: 10.1038/mt.2013.282. Epub 2013 Dec 12.
Pompe disease is an autosomal recessive disorder caused by mutations in the acid-α glucosidase (GAA) gene. Lingual dysfunction is prominent but does not respond to conventional enzyme replacement therapy (ERT). Using Pompe (Gaa(-/-)) mice, we tested the hypothesis that intralingual delivery of viral vectors encoding GAA results in GAA expression and glycogen clearance in both tongue myofibers and hypoglossal (XII) motoneurons. An intralingual injection of an adeno-associated virus (AAV) vector encoding GAA (serotypes 1 or 9; 1 × 10(11) vector genomes, CMV promoter) was performed in 2-month-old Gaa(-/-) mice, and tissues were harvested 4 months later. Both serotypes robustly transduced tongue myofibers with histological confirmation of GAA expression (immunochemistry) and glycogen clearance (Period acid-Schiff stain). Both vectors also led to medullary transgene expression. GAA-positive motoneurons did not show the histopathologic features which are typical in Pompe disease and animal models. Intralingual injection with the AAV9 vector resulted in approximately threefold more GAA-positive XII motoneurons (P < 0.02 versus AAV1); the AAV9 group also gained more body weight over the course of the study (P < 0.05 versus AAV1 and sham). We conclude that intralingual injection of AAV1 or AAV9 drives persistent GAA expression in tongue myofibers and motoneurons, but AAV9 may more effectively target motoneurons.
庞贝病是一种常染色体隐性疾病,由酸性α-葡萄糖苷酶(GAA)基因突变引起。舌功能障碍较为突出,但对传统的酶替代疗法(ERT)无反应。我们利用庞贝病(Gaa(-/-))小鼠,检验了以下假说:向舌内递送编码GAA的病毒载体可导致舌肌纤维和舌下(XII)运动神经元中GAA表达及糖原清除。对2月龄的Gaa(-/-)小鼠进行舌内注射编码GAA的腺相关病毒(AAV)载体(血清型1或9;1×10(11)载体基因组,CMV启动子),4个月后采集组织。两种血清型均能有效地转导舌肌纤维,通过免疫化学证实GAA表达,过碘酸希夫染色证实糖原清除。两种载体还导致延髓转基因表达。GAA阳性运动神经元未表现出庞贝病及动物模型中典型的组织病理学特征。舌内注射AAV9载体导致GAA阳性XII运动神经元数量增加约三倍(与AAV1相比,P < 0.02);在研究过程中,AAV9组体重增加也更多(与AAV1和假注射组相比,P < 0.05)。我们得出结论,舌内注射AAV1或AAV9可驱动舌肌纤维和运动神经元中持续的GAA表达,但AAV9可能更有效地靶向运动神经元。