Dorn Gabriele, Patel Sadhana, Wotherspoon Glen, Hemmings-Mieszczak Maja, Barclay Jane, Natt Francois J C, Martin Pierre, Bevan Stuart, Fox Alyson, Ganju Pam, Wishart William, Hall Jonathan
Functional Genomics, Novartis Institutes for Biomedical Research, Novartis Pharma AG, 4002 Basel, Switzerland.
Nucleic Acids Res. 2004 Mar 16;32(5):e49. doi: 10.1093/nar/gnh044.
Double stranded, short interfering RNAs (siRNA) of 21-22 nt length initiate a sequence-specific, post-trancriptional gene silencing in animals and plants known as RNA interference (RNAi). Here we show that RNAi can block a pathophysiological pain response and provide relief from neuropathic pain in a rat disease model by down regulating an endogenous, neuronally expressed gene. Rats, intrathecally infused with a 21 nt siRNA perfectly complementary to the pain-related cation-channel P2X3, showed diminished pain responses compared to missense (MS) siRNA-treated and untreated controls in models of both agonist-evoked pain and chronic neuropathic pain. This form of delivery caused no adverse effects in any of the animals receiving P2X3 siRNA, MS siRNA or vehicle. Molecular analysis of tissues revealed that P2X3 mRNA expressed in dorsal root ganglia, and P2X3 protein translocated into the dorsal horn of the spinal cord, were significantly diminished. These observations open a path toward use of siRNA as a genetic tool for drug target validation in the mammalian central nervous system, as well as for proof of concept studies and as therapeutic agents in man.
长度为21 - 22个核苷酸的双链短干扰RNA（siRNA）在动植物中引发一种序列特异性的转录后基因沉默，即RNA干扰（RNAi）。在此我们表明，RNAi可通过下调内源性神经元表达基因，阻断病理生理疼痛反应，并在大鼠疾病模型中缓解神经性疼痛。在激动剂诱发疼痛和慢性神经性疼痛模型中，鞘内注射与疼痛相关阳离子通道P2X3完全互补的21个核苷酸siRNA的大鼠，与错义（MS）siRNA处理组和未处理的对照组相比，疼痛反应减弱。这种给药方式在接受P2X3 siRNA、MS siRNA或赋形剂的任何动物中均未产生不良反应。组织的分子分析显示，背根神经节中表达的P2X3 mRNA以及转运至脊髓背角的P2X3蛋白均显著减少。这些观察结果为将siRNA用作哺乳动物中枢神经系统药物靶点验证的遗传工具、概念验证研究以及人类治疗药物开辟了一条道路。
