Franco Luis M, Sun Baodong, Yang Xiaoyi, Bird Andrew, Zhang Haoyue, Schneider Ayn, Brown Talmage, Young Sarah P, Clay Timothy M, Amalfitano Andrea, Chen Y T, Koeberl Dwight D
Division of Medical Genetics, Department of Pediatrics, Duke University Medical Center, Durham, NC 27710, USA.
Mol Ther. 2005 Nov;12(5):876-84. doi: 10.1016/j.ymthe.2005.04.024. Epub 2005 Jul 6.
Glycogen storage disease type II (GSD-II; Pompe disease) is caused by a deficiency of acid alpha-glucosidase (GAA; acid maltase) and manifests as muscle weakness, hypertrophic cardiomyopathy, and respiratory failure. Adeno-associated virus vectors containing either a liver-specific promoter (LSP) (AAV-LSPhGAApA) or a hybrid CB promoter (AAV-CBhGAApA) to drive human GAA expression were pseudotyped as AAV8 and administered to immunocompetent GAA-knockout mice. Secreted hGAA was detectable in plasma between 1 day and 12 weeks postadministration with AAV-LSPhGAApA and only from 1 to 8 days postadministration for AAV-CBGAApA. No anti-GAA antibodies were detected in response to AAV-LSPhGAApA (<1:200), whereas AAV-CBhGAApA provoked an escalating antibody response starting 2 weeks postadministration. The LSP drove approximately 60-fold higher GAA expression than the CB promoter in the liver by 12 weeks following vector administration. Furthermore, the detected cellular immunity was provoked by AAV-CBhGAApA, as detected by ELISpot and CD4+/CD8+ lymphocyte immunodetection. GAA activity was increased to higher than normal and glycogen content was reduced to essentially normal levels in the heart and skeletal muscle following administration of AAV-LSPhGAApA. Therefore, liver-restricted GAA expression with an AAV vector evaded immunity and enhanced efficacy in GSD-II mice.
II型糖原贮积病(GSD-II;庞贝病)由酸性α-葡萄糖苷酶(GAA;酸性麦芽糖酶)缺乏引起,表现为肌肉无力、肥厚型心肌病和呼吸衰竭。将含有肝脏特异性启动子(LSP)(AAV-LSPhGAApA)或杂交CB启动子(AAV-CBhGAApA)以驱动人GAA表达的腺相关病毒载体伪型化为AAV8,并给予具有免疫活性的GAA基因敲除小鼠。给予AAV-LSPhGAApA后1天至12周血浆中可检测到分泌的hGAA,而给予AAV-CBGAApA后仅在给药后1至8天可检测到。对AAV-LSPhGAApA未检测到抗GAA抗体(<1:200),而AAV-CBhGAApA在给药后2周开始引发不断升级的抗体反应。载体给药12周后,LSP在肝脏中驱动的GAA表达比CB启动子高约60倍。此外,通过ELISpot和CD4+/CD8+淋巴细胞免疫检测发现,AAV-CBhGAApA引发了检测到的细胞免疫。给予AAV-LSPhGAApA后,心脏和骨骼肌中的GAA活性增加至高于正常水平,糖原含量降至基本正常水平。因此,用腺相关病毒载体进行肝脏限制性GAA表达可避免免疫反应并提高GSD-II小鼠的疗效。