Muscle-specific promoters may be necessary for adeno-associated virus-mediated gene transfer in the treatment of muscular dystrophies
Muscle-specific promoters may be necessary for adeno-associated virus-mediated gene transfer in the treatment of muscular dystrophies
复制标题
DOI：
10.1089/104303401750061267
复制
发表时间：
2001-01-20
影响因子：
4.2
通讯作者：
Sweeney, HL
中科院分区：
文献类型：
--
作者：
Cordier, L;Gao, GP;Sweeney, HL
重组腺相关病毒（rAAV）载体允许高效的基因转移和在肌肉中的表达；因此，raav代表了肌营养不良症的潜在基因治疗载体。为了进一步的研究，我们使用了小鼠肌营养不良模型（gsg（-/-）小鼠），缺失了肌营养不良蛋白-糖蛋白复合物的亚基γ -肌聚糖。Gsg（-/-）小鼠出现进行性营养不良，代表严重的人类表型疾病。我们之前在gsg（-/-）小鼠中直接肌肉注射了一种重组AAV载体（AAV. dmck . gsg），该载体携带由肌肉特异性启动子（肌酸激酶的截断版本）驱动的γ -肌聚糖cDNA，在肌纤维中高水平和稳定地表达了γ -肌聚糖。当使用rAAV载体，在CMV启动子（AAV.CMV）下表达高免疫原性产物γ -半乳糖苷酶时，观察到较低水平的转基因表达，并与对γ -肌聚糖的体液反应有关。LacZ)，我们测量了gsg（-/-）小鼠肌肉注射后对转基因的强烈细胞和体液免疫反应。这项研究表明，限制转基因在肌肉中的表达是治疗肌肉萎缩症的一个重要标准，并将有助于设计基因治疗方案。
Recombinant adeno-associated virus (rAAV) vectors allow efficient gene transfer and expression in the muscle; therefore, rAAVs represent a potential gene therapy vector for muscular dystrophies, For further investigations, we used a mouse muscular dystrophy model (gsg(-/-) mice) gamma -sarcoglycan, a subunit of the dystrophin-glycoprotein complex, is missing. gsg(-/-) mice develop progressive dystrophy representative of a severe human phenotype disease. We previously showed high levels and stable expression of gamma -sarcoglycan in myofibers after direct muscle injection into gsg(-/-) mice of a recombinant AAV vector (AAV.dMCK.gSG) carrying the gamma -sarcoglycan cDNA driven by a muscle-specific promoter (truncated version of muscle creatine kinase), Here, we show that when gamma -sarcoglycan expression is driven by the ubiquitous cytomegalovirus (CMV) promoter (AAV.CMV.gSG), lower levels of transgene expression are observed and are associated with a humoral response to gamma -sarcoglycan, When using an rAAV vector, expressing the highly immunogenic product gamma -galactosidase under the CMV promoter (AAV.CMV.LacZ), we measured a strong cellular and humoral immune response to the transgene after intramuscular injection into gsg(-/-) mice. This study suggests that restriction of transgene expression to the muscle is an important criterion for the treatment of muscular dystrophies and will aid in the design of protocols for gene therapy.