Efficient and persistent splice switching by systemically delivered LNA oligonucleotides in mice
Efficient and persistent splice switching by systemically delivered LNA oligonucleotides in mice
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DOI：
10.1016/j.ymthe.2006.05.017
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发表时间：
2006-10-01
影响因子：
12.4
通讯作者：
Kole, Ryszard
中科院分区：
文献类型：
--
作者：
Roberts, Jennifer;Palma, Enzo;Kole, Ryszard
发现锁核酸（LNA）寡聚体在转基因小鼠体内调节可变剪接的能力方面非常有效，所述转基因小鼠普遍表达含有异常剪接的β-珠蛋白内含子（IVS 2 -654）的经修饰的EGFP前mRNA。腹膜内注射后，靶向EGFP-654前体mRNA中异常5'剪接位点的剪接转换寡核苷酸LNA SSO-654纠正了异常剪接，并主要在肝脏、结肠和小肠中增加了修复的EGFP mRNA的产生。在心脏、肺或肾脏中检测到很少或没有影响，在连续四次每日注射后，大部分寡核苷酸分布在这些器官中。在肝脏中，LNA SSO-654的EC 50为3 mg/kg，比其2 '-O-甲基同系物的效力高约17倍。此外，在肝脏、结肠和小肠中，50 mg/kg的口服剂量导致可检测水平的剪接转换。以25 mg/kg剂量每日注射4次的影响持续长达29天，但未导致肝毒性。结果表明，LNA骨架赋予寡核苷酸的序列和器官特异性功能生物分布，并且这些有效的化合物具有成为可通过剪接操作治疗的疾病的安全和长效调节剂的潜力。
Locked nucleic acid (LNA) oligomers were found to be very effective in their ability to modulate alternative splicing in vivo in transgenic mice that ubiquitously express a modified EGFP pre-mRNA containing an aberrantly spliced beta-globin intron (IVS2-654). Following intraperitoneal injections, the splice-switching oligonucleotide LNA SSO-654 targeted to the aberrant 5' splice site in EGFP-654 pre-mRNA corrected aberrant splicing and increased production of repaired EGFP mRNA mainly in the liver, colon, and small intestine. Little or no effect was detected in heart, lung, or kidney, the organ where most of the oligonucleotide was distributed after four consecutive daily injections. In the liver, LNA SSO-654 had an EC50 of 3 mg/kg, approximately 17-fold more potent than its 2'-O-methyl congener. Moreover, in the liver, colon, and small intestine oral doses of 50 mg/kg resulted in detectable levels of splice switching. The effects of four daily injections at 25 mg/kg persisted for up to 29 days but did not result in liver toxicity. The results indicate that the LNA backbone confers sequence- and organ-specific functional biodistribution of the oligonucleotides and that these potent compounds have the potential to be safe and long-acting modulators of diseases treatable by splicing manipulation.