2'-O-甲氧乙基反义寡核苷酸对杜兴肌肉萎缩症治疗的研究

Duchenne Muscular Dystrophy (DMD) is a lethal neuromuscular disorder caused by the absence of dystrophin protein. In our present study, we evaluated a new antisense oligonucleotide (AO) chemistry-2'-O-Methoxyethyl (2'-O-MOE) in a DMD animal model in vitro and in vivo, and wish to provide a candidate AO drug for treating DMD patients. Methods: RT-PCR was utilized for detecting the exon skipping efficiency after in vitro transfection with different lengths and different backbone modifications of 2'-O-MOE AOs in H2K mdx cells. Selected 2'-O-MOE AOs were injected into tibialis anterior (TA) muscle of adult mdx mice after in vitro evaluation. Treated TA muscles were harvested two weeks post-injection and assayed by immunohistochemistry, RT-PCR and Western blot to measure the exon skipping efficiency and the level of dystrophin restoration. Results: Three tested 2'-O-MOE AOs show significantly higher level of exon skipping efficiency than that of 2'OmePS, which is used in current clinical trials, based on in vitro study. In line with in vitro data, local intramuscular study shows the same pattern with 2'-O-MOE AOs inducing higher level of exon skipping and dystrophin expression than that of 2'OmePS. Conclusions: 2'-O-MOE AOs are effective in inducing the expression of dystophin in in vitro and TA muscles of DMD animal models.