Reijonen Sami, Putkonen Noora, Nørremølle Anne, Lindholm Dan, Korhonen Laura
Medical Research Institute Minerva, Biomedicum Helsinki, Haartmaninkatu 8, Helsinki, Finland.
Exp Cell Res. 2008 Mar 10;314(5):950-60. doi: 10.1016/j.yexcr.2007.12.025. Epub 2008 Jan 14.
Accumulation of abnormal proteins occurs in many neurodegenerative diseases including Huntington's disease (HD). However, the precise role of protein aggregation in neuronal cell death remains unclear. We show here that the expression of N-terminal huntingtin proteins with expanded polyglutamine (polyQ) repeats causes cell death in neuronal PC6.3 cell that involves endoplasmic reticulum (ER) stress. These mutant huntingtin fragment proteins elevated Bip, an ER chaperone, and increased Chop and the phosphorylation of c-Jun-N-terminal kinase (JNK) that are involved in cell death regulation. Caspase-12, residing in the ER, was cleaved in mutant huntingtin expressing cells, as was caspase-3 mediating cell death. In contrast, cytochrome-c or apoptosis inducing factor (AIF) was not released from mitochondria after the expression of these proteins. Treatment with salubrinal that inhibits ER stress counteracted cell death and reduced protein aggregations in the PC6.3 cells caused by the mutant huntingtin fragment proteins. Salubrinal upregulated Bip, reduced cleavage of caspase-12 and increased the phosphorylation of eukaryotic translation initiation factor-2 subunit-alpha (eIF2alpha) that are neuroprotective. These results show that N-terminal mutant huntingtin proteins activate cellular pathways linked to ER stress, and that inhibition of ER stress by salubrinal increases cell survival. The data suggests that compounds targeting ER stress may be considered in designing novel approaches for treatment of HD and possibly other polyQ diseases.
包括亨廷顿舞蹈症(HD)在内的许多神经退行性疾病中都会出现异常蛋白质的积累。然而,蛋白质聚集在神经元细胞死亡中的确切作用仍不清楚。我们在此表明,具有扩展的聚谷氨酰胺(polyQ)重复序列的N端亨廷顿蛋白的表达会导致神经元PC6.3细胞死亡,这一过程涉及内质网(ER)应激。这些突变的亨廷顿片段蛋白提高了内质网伴侣蛋白Bip的水平,增加了参与细胞死亡调节的Chop以及c-Jun氨基末端激酶(JNK)的磷酸化。位于内质网的半胱天冬酶-12在表达突变亨廷顿蛋白的细胞中被切割,介导细胞死亡的半胱天冬酶-3也是如此。相比之下,在这些蛋白表达后,细胞色素c或凋亡诱导因子(AIF)并未从线粒体中释放。用抑制内质网应激的水杨酰胺进行处理可抵消细胞死亡,并减少由突变亨廷顿片段蛋白导致的PC6.3细胞中的蛋白质聚集。水杨酰胺上调了Bip,减少了半胱天冬酶-12的切割,并增加了具有神经保护作用的真核翻译起始因子2α亚基(eIF2α)的磷酸化。这些结果表明,N端突变亨廷顿蛋白激活了与内质网应激相关的细胞途径,并且水杨酰胺对内质网应激的抑制增加了细胞存活。数据表明,在设计治疗HD及可能的其他polyQ疾病的新方法时,可以考虑针对内质网应激的化合物。