Tang Qingquan, Li Baojian, Woodle Martin, Lu Patrick Y
OriGene Technologies Inc, Rockville, MD, USA.
Methods Mol Biol. 2008;442:139-58. doi: 10.1007/978-1-59745-191-8_11.
Containment of the SARS coronavirus (SCV) outbreak was accompanied by the rapid characterization of this new pathogen's genome sequence in 2003, encouraging the development of anti-SCV therapeutics using short interfering RNA (siRNA) inhibitors. A pair of siRNA duplexes identified as potent SCV inhibitors in vitro was evaluated for in vivo efficacy and safety in a rhesus macaque SARS model using intranasal administration with clinical viable delivery carrier in three dosing regimens. Observations of SCV-induced SARS-like symptoms, measurements of SCV RNA presence in the respiratory tract, microscopic inspections of lung histopathology, and immunohistochemistry sections from 21 tested macaques consistently demonstrated siRNA-mediated anti-SCV activity. The prophylactic and therapeutic efficacies resulted in relief of animals from SCV infection-induced fever, diminished SCV in upper airway and lung alveoli, and milder acute diffuse alveoli damage (DAD). The dosages of siRNA used, 10 to 40 mg/kg, did not show any sign of siRNA-induced toxicity. These results support that a clinical investigation of this anti-SARS siRNA therapeutic agent is warranted. The study also illustrates the capability of siRNA to enable a massive reduction in development time for novel targeted therapeutic agents. We detail a representative example of large-mammal siRNA use.
2003年，严重急性呼吸综合征冠状病毒（SCV）疫情的控制伴随着对这种新型病原体基因组序列的快速鉴定，这推动了使用小干扰RNA（siRNA）抑制剂开发抗SCV疗法。在恒河猴SARS模型中，采用三种给药方案，通过鼻腔给药并结合临床可行的递送载体，对一对在体外被鉴定为有效SCV抑制剂的siRNA双链体进行了体内疗效和安全性评估。对21只受试猕猴的SCV诱导的类SARS症状观察、呼吸道中SCV RNA存在情况的测量、肺组织病理学的显微镜检查以及免疫组织化学切片，均一致证明了siRNA介导的抗SCV活性。预防和治疗效果使动物从SCV感染引起的发热中得到缓解，上呼吸道和肺泡中的SCV减少，急性弥漫性肺泡损伤（DAD）减轻。所使用的siRNA剂量为10至40 mg/kg，未显示出任何siRNA诱导毒性的迹象。这些结果支持对这种抗SARS siRNA治疗剂进行临床研究。该研究还说明了siRNA能够大幅缩短新型靶向治疗剂的研发时间。我们详细介绍了一个使用大哺乳动物siRNA的代表性例子。
