Rigamonti Dorotea, Bolognini Daniele, Mutti Cesare, Zuccato Chiara, Tartari Marzia, Sola Francesco, Valenza Marta, Kazantsev Aleksey G, Cattaneo Elena
Centre for Stem Cell Research and Department of Pharmacological Sciences, University of Milan, Via Balzaretti 9, Milan 20133, Italy.
J Biol Chem. 2007 Aug 24;282(34):24554-62. doi: 10.1074/jbc.M609885200. Epub 2007 Jun 12.
Increased levels of the repressor element 1/neuron restrictive silencer element (RE1/NRSE) silencing activity promoter, and a consequent reduction in the transcription of many RE1/NRSE-bearing neuronal genes, including brain-derived neurotrophic factor (BDNF), have been demonstrated in Huntington disease (HD) and represent one possible effector of its selective neuronal vulnerability. Restoring the expression levels of neuronal genes in diseased neurons therefore seems to be an attractive therapeutic approach. To this end, we have developed a cell-based reporter assay for monitoring RE1/NRSE silencing activity and validated it by genetically inactivating the RE1/NRSE or pharmacologically stimulating global transcription. In a pilot compound screen, we identified three closely related structural analogues that up-regulate reporter expression at low nanomolar concentrations, and follow-up studies have shown that they efficaciously increase endogenous BDNF levels in HD cells. Moreover, one of the compounds increases the viability of HD cells. Our findings suggest a new avenue for the development of drugs for HD and other neurodegenerative disorders based on the pharmacological up-regulation of the production of the neuronal survival factor BDNF and of other RE1/NRSE-regulated neuronal genes.
在亨廷顿舞蹈病(HD)中,阻遏元件1/神经元限制性沉默元件(RE1/NRSE)沉默活性启动子水平升高,导致许多含有RE1/NRSE的神经元基因转录减少,包括脑源性神经营养因子(BDNF),这代表了其选择性神经元易损性的一种可能效应器。因此,恢复患病神经元中神经元基因的表达水平似乎是一种有吸引力的治疗方法。为此,我们开发了一种基于细胞的报告基因检测方法来监测RE1/NRSE沉默活性,并通过基因灭活RE1/NRSE或药理学刺激全局转录对其进行了验证。在一次先导化合物筛选中,我们鉴定出三种密切相关的结构类似物,它们在低纳摩尔浓度下上调报告基因表达,后续研究表明它们能有效提高HD细胞中内源性BDNF水平。此外,其中一种化合物可提高HD细胞的活力。我们的研究结果为基于药理学上调神经元存活因子BDNF及其他RE1/NRSE调控的神经元基因的产生来开发治疗HD和其他神经退行性疾病的药物开辟了一条新途径。