Dept. of Biomedical Engineering, University of Minnesota, Minneapolis, MN, 55455, USA.
Dept. of Medicinal Chemistry, University of Minnesota, Minneapolis, MN, 55455, USA.
Mol Neurobiol. 2023 Oct;60(10):6133-6144. doi: 10.1007/s12035-023-03417-5. Epub 2023 Jul 10.
Proteasomal degradation of intrinsically disordered proteins, such as tau, is a critical component of proteostasis in both aging and neurodegenerative diseases. In this study, we investigated proteasomal activation by MK886 (MK). We previously identified MK as a lead compound capable of modulating tau oligomerization in a cellular FRET assay and rescuing P301L tau-induced cytotoxicity. We first confirmed robust proteasomal activation by MK using 20S proteasomal assays and a cellular proteasomal tau-GFP cleavage assay. We then show that MK treatment can significantly rescue tau-induced neurite pathology in differentiated SHSY5Y neurospheres. Due to this compelling result, we designed a series of seven MK analogs to determine if proteasomal activity is sensitive to structural permutations. Using the proteasome as the primary MOA, we examined tau aggregation, neurite outgrowth, inflammation, and autophagy assays to identify two essential substituents of MK that are required for compound activity: (1) removal of the N-chlorobenzyl group from MK negated both proteasomal and autophagic activity and reduced neurite outgrowth; and (2) removal of the indole-5-isopropyl group significantly improved neurite outgrowth and autophagy activity but reduced its anti-inflammatory capacity. Overall, our results suggest that the combination of proteasomal/autophagic stimulation and anti-inflammatory properties of MK and its derivatives can decrease tau-tau interactions and help rebalance dysfunctional proteostasis. Further development of MK to optimize its proteasomal, autophagic, and anti-inflammatory targets may lead to a novel therapeutic that would be beneficial in aging and neurodegenerative diseases.
蛋白酶体降解内在无序蛋白质(如 tau)是衰老和神经退行性疾病中蛋白质稳态的关键组成部分。在这项研究中,我们研究了 MK886(MK)对蛋白酶体的激活作用。我们之前发现 MK 是一种能够在细胞 FRET 测定中调节 tau 寡聚化并挽救 P301L tau 诱导的细胞毒性的先导化合物。我们首先使用 20S 蛋白酶体测定和细胞蛋白酶体 tau-GFP 切割测定证实了 MK 对蛋白酶体的强大激活作用。然后,我们表明 MK 处理可以显著挽救分化的 SHSY5Y 神经球中的 tau 诱导的神经突病理学。由于这一引人注目的结果,我们设计了一系列七种 MK 类似物,以确定蛋白酶体活性是否对结构变化敏感。我们使用蛋白酶体作为主要的作用机制,检查 tau 聚集、神经突生长、炎症和自噬测定,以确定 MK 活性所必需的两个关键取代基:(1)从 MK 中去除 N-氯苯甲基基团会使蛋白酶体和自噬活性均失活,并减少神经突生长;(2)去除吲哚-5-异丙基基团可显著改善神经突生长和自噬活性,但降低其抗炎能力。总体而言,我们的结果表明,MK 及其衍生物的蛋白酶体/自噬刺激和抗炎特性的结合可以减少 tau-tau 相互作用,并有助于重新平衡功能失调的蛋白质稳态。进一步开发 MK 以优化其蛋白酶体、自噬和抗炎靶点可能会导致一种新的治疗方法,对衰老和神经退行性疾病有益。