Stability study of unmodified siRNA and relevance to clinical use.
Stability study of unmodified siRNA and relevance to clinical use.
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DOI：
10.1089/oli.2008.0149
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发表时间：
2008-12
期刊：
影响因子：
--
通讯作者：
Kaspar RL
中科院分区：
文献类型：
--
作者：
Hickerson RP;Vlassov AV;Wang Q;Leake D;Ilves H;Gonzalez-Gonzalez E;Contag CH;Johnston BH;Kaspar RL
RNA干扰为开发各种疾病的治疗剂提供了巨大的潜力。为了评估siRNAs在与临床使用相关的条件下的稳定性，特别强调局部递送考虑，对三种不同的未经修饰的siRNAs进行了研究。结果表明，通过非变性聚丙烯酰胺凝胶电泳法和功能活性测定，重复冻融循环、延长孵化时间(在21℃下超过1年)和高温(高达95℃)下较短的孵育时间对siRNA的完整性没有任何影响。在37℃下接触毛发或皮肤后也没有观察到降解，然而，在37℃下孵育在胎牛或人血清中会导致降解和活性丧失。因此，血液中的siRNA可能被灭活，从而限制了全身暴露。有趣的是，部分降解(通过凝胶电泳法观察到)并不总是与活性丧失相关，这表明部分降解的siRNA保留了全部功能活性。为了证明未修饰的siRNA的功能活性，将绿色荧光蛋白特异的抑制剂注射到足垫中，并在转基因报告鼠模型中显示抑制预先存在的绿色荧光蛋白的表达。综上所述，这些数据表明未经修饰的siRNA是可行的治疗候选药物。
RNA interference offers enormous potential to develop therapeutic agents for a variety of diseases. To assess the stability of siRNAs under conditions relevant to clinical use with particular emphasis on topical delivery considerations, a study of three different unmodified siRNAs was performed. The results indicate that neither repeated freeze/thaw cycles, extended incubations (over 1 year at 21°C), nor shorter incubations at high temperatures (up to 95°C) have any effect on siRNA integrity as measured by nondenaturing polyacrylamide gel electrophoresis and functional activity assays. Degradation was also not observed following exposure to hair or skin at 37°C. However, incubation in fetal bovine or human sera at 37°C led to degradation and loss of activity. Therefore, siRNA in the bloodstream is likely inactivated, thereby limiting systemic exposure. Interestingly, partial degradation (observed by gel electrophoresis) did not always correlate with loss of activity, suggesting that partially degraded siRNAs retain full functional activity. To demonstrate the functional activity of unmodified siRNA, EGFP-specific inhibitors were injected into footpads and shown to inhibit preexisting EGFP expression in a transgenic reporter mouse model. Taken together, these data indicate that unmodified siRNAs are viable therapeutic candidates.
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影响因子：
2.9
作者：
Braasch, DA;Jensen, S;Corey, DR
通讯作者：
Corey, DR
影响因子：
64.8
作者：
Soutschek, J;Akinc, A;Vornlocher, HP
通讯作者：
Vornlocher, HP
影响因子：
6.4
作者：
Haupenthal, Joerg;Baehr, Christina;Piiper, Albrecht
通讯作者：
Piiper, Albrecht
影响因子：
4.5
作者：
Layzer, JM;McCaffrey, AP;Sullenger, BA
通讯作者：
Sullenger, BA
DOI：
10.1038/nrd2310
发表时间：
2007-06
期刊：
Nature reviews. Drug discovery
影响因子：
--
作者：
de Fougerolles A;Vornlocher HP;Maraganore J;Lieberman J
通讯作者：
Lieberman J