Daude Nathalie, Marella Mathieu, Chabry Joelle
Institut de Pharmacologie Moléculaire et Cellulaire, Unité Mixte de Recherche 6097, Centre National de la Recherche Scientifique. 660, route des lucioles, 06560 Valbonne, France.
J Cell Sci. 2003 Jul 1;116(Pt 13):2775-9. doi: 10.1242/jcs.00494. Epub 2003 May 20.
Development of transmissible spongiform encephalopathies (TSEs) pathogenesis requires the presence of both the normal host prion protein (PrP-sen) and the abnormal pathological proteinase-K resistant isoform (PrP-res). PrP-res forms highly insoluble aggregates, with self-perpetuating properties, by binding and converting PrP-sen molecules into a likeness of themselves. In the present report, we show that small interfering RNA (siRNA) duplexes trigger specific Prnp gene silencing in scrapie-infected neuroblastoma cells. A non-passaged, scrapie-infected culture transfected with siRNA duplexes is depleted of PrP-sen and rapidly loses its PrP-res content. The use of different murine-adapted scrapie strains and host cells did not influence the siRNA-induced gene silencing efficiency. More than 80% of transfected cells were positive for the presence of fluorescein-labeled siRNA duplexes. No cytotoxicity associated with the use of siRNA was observed during the time course of these experiments. Despite a transient abrogation of PrP-res accumulation, our results suggest that the use of siRNA may provide a new and promising therapeutic approach against prion diseases.
传染性海绵状脑病（TSEs）发病机制的发展需要正常宿主朊病毒蛋白（PrP-sen）和异常的抗蛋白酶K病理异构体（PrP-res）同时存在。PrP-res通过结合并将PrP-sen分子转化为自身的类似物，形成高度不溶性聚集体，并具有自我延续的特性。在本报告中，我们表明小干扰RNA（siRNA）双链体可在感染羊瘙痒病的神经母细胞瘤细胞中引发特异性Prnp基因沉默。用siRNA双链体转染的未经传代的感染羊瘙痒病的培养物中PrP-sen减少，并迅速失去其PrP-res含量。使用不同的鼠适应羊瘙痒病株和宿主细胞并不影响siRNA诱导的基因沉默效率。超过80%的转染细胞中存在荧光素标记的siRNA双链体呈阳性。在这些实验的时间过程中，未观察到与使用siRNA相关的细胞毒性。尽管PrP-res积累暂时被消除，但我们的结果表明，使用siRNA可能为抗朊病毒疾病提供一种新的、有前景的治疗方法。
