Instituto de Investigación Médica M y M Ferreyra, INIMEC-CONICET, Universidad Nacional de Córdoba, Córdoba 5016, Argentina.
Int J Mol Sci. 2023 Apr 13;24(8):7209. doi: 10.3390/ijms24087209.
The Unfolded protein response (UPR), triggered by stress in the endoplasmic reticulum (ER), is a key driver of neurodegenerative diseases. GM2 gangliosidosis, which includes Tay-Sachs and Sandhoff disease, is caused by an accumulation of GM2, mainly in the brain, that leads to progressive neurodegeneration. Previously, we demonstrated in a cellular model of GM2 gangliosidosis that PERK, a UPR sensor, contributes to neuronal death. There is currently no approved treatment for these disorders. Chemical chaperones, such as ursodeoxycholic acid (UDCA), have been found to alleviate ER stress in cell and animal models. UDCA's ability to move across the blood-brain barrier makes it interesting as a therapeutic tool. Here, we found that UDCA significantly diminished the neurite atrophy induced by GM2 accumulation in primary neuron cultures. It also decreased the up-regulation of pro-apoptotic CHOP, a downstream PERK-signaling component. To explore its potential mechanisms of action, in vitro kinase assays and crosslinking experiments were performed with different variants of recombinant protein PERK, either in solution or in reconstituted liposomes. The results suggest a direct interaction between UDCA and the cytosolic domain of PERK, which promotes kinase phosphorylation and dimerization.
未折叠蛋白反应(UPR)是内质网(ER)应激触发的一种关键的神经退行性疾病驱动因素。GM2 神经节苷脂贮积症包括泰萨二氏症和桑德霍夫病,是由于 GM2 的积累,主要在大脑中,导致进行性神经退行性变。以前,我们在 GM2 神经节苷脂贮积症的细胞模型中证明,UPR 传感器 PERK 有助于神经元死亡。目前,这些疾病还没有批准的治疗方法。化学伴侣,如熊去氧胆酸(UDCA),已被发现可减轻细胞和动物模型中的 ER 应激。UDCA 能够穿过血脑屏障,这使其成为一种有前途的治疗工具。在这里,我们发现 UDCA 可显著减少 GM2 积累在原代神经元培养物中诱导的神经突起萎缩。它还降低了 PERK 信号下游促凋亡 CHOP 的上调。为了探索其潜在的作用机制,我们进行了体外激酶测定和用不同变体的重组蛋白 PERK 进行交联实验,无论是在溶液中还是在重建的脂质体中。结果表明 UDCA 与 PERK 的细胞质结构域之间存在直接相互作用,这促进了激酶磷酸化和二聚化。