Department of Demyelinating Disease and Aging, National Institute of Neuroscience, NCNP, Kodaira, Tokyo 187-8502, Japan; Department of Neurology, Faculty of Medicine, University of Tsukuba, Tsukuba, Ibaraki 305-8575, Japan.
Department of Demyelinating Disease and Aging, National Institute of Neuroscience, NCNP, Kodaira, Tokyo 187-8502, Japan.
Neuroscience. 2019 Mar 15;402:11-22. doi: 10.1016/j.neuroscience.2019.01.010. Epub 2019 Jan 21.
Abnormalities of the autophagy-lysosomal pathway (ALP) have been implicated in the pathology of Alzheimer's disease (AD). Activation of TFEB (transcription factor EB), a master regulator of the ALP, leads to ALP facilitation. The present study sought to clarify whether TFEB-mediated ALP facilitation influences the process of amyloid β-protein (Aβ) generation in neurons. TFEB was overexpressed in mature rat primary cortical neurons via recombinant adenoviruses, without (basal conditions) or with co-overexpression of wild-type amyloid precursor protein (APP) or its β-C-terminal fragment (β-CTF). We confirmed that TFEB overexpression upregulated the lysosomal proteins, cathepsin D and LAMP-1. In TFEB-expressing neurons, protein levels of ADAM10 were profoundly increased, whereas those of APP, BACE1, or γ-secretase complex proteins were unaffected. However, TFEB did not affect ADAM10 mRNA levels. TFEB overexpression had different effects on Aβ production depending on the expression level of APP or β-CTF: TFEB slightly decreased Aβ secretion under basal conditions; clearly increased α-CTF levels and marginally increased β-CTF levels with modest increases in secreted Aβ in APP-expressing neurons; and caused a remarkable increase in β-CTF levels with a significant increase in secreted Aβ in β-CTF-expressing neurons. Inhibition of proteasomes, but not lysosomes, markedly increased β-CTF levels in β-CTF-expressing neurons. These results collectively indicate that TFEB modulates Aβ production not only by increasing α-secretase processing of APP through ADAM10 upregulation but also by augmenting β-CTF levels possibly via altered proteasome-mediated catabolism. Thus, TFEB-mediated ALP enhancement appears to have dual, but opposite, effects on Aβ production in neurons.
自噬溶酶体途径 (ALP) 的异常与阿尔茨海默病 (AD) 的病理学有关。TFEB(转录因子 EB)的激活是 ALP 的主要调节剂,可导致 ALP 促进。本研究旨在阐明 TFEB 介导的 ALP 促进是否会影响神经元中淀粉样 β-蛋白 (Aβ) 的产生过程。通过重组腺病毒在成熟的大鼠原代皮质神经元中过表达 TFEB,在没有(基础条件)或与野生型淀粉样前体蛋白 (APP) 或其 β-C 末端片段 (β-CTF) 共过表达的情况下。我们证实 TFEB 过表达上调了溶酶体蛋白组织蛋白酶 D 和 LAMP-1。在 TFEB 表达的神经元中,ADAM10 的蛋白水平显著增加,而 APP、BACE1 或 γ-分泌酶复合物蛋白的水平不受影响。然而,TFEB 不影响 ADAM10 mRNA 水平。TFEB 过表达对 Aβ 产生的影响取决于 APP 或 β-CTF 的表达水平:TFEB 在基础条件下略微减少 Aβ 的分泌;在 APP 表达的神经元中,明显增加 α-CTF 水平和轻微增加 β-CTF 水平,并适度增加分泌的 Aβ;在 β-CTF 表达的神经元中,导致 β-CTF 水平显著增加,并显著增加分泌的 Aβ。蛋白酶体抑制剂,但不是溶酶体抑制剂,明显增加了β-CTF 在β-CTF 表达的神经元中的水平。这些结果共同表明,TFEB 不仅通过上调 ADAM10 增加 APP 的 α-分泌酶加工来调节 Aβ 的产生,而且还通过改变蛋白酶体介导的分解代谢来增加β-CTF 水平,从而对 Aβ 的产生产生双重但相反的影响。因此,TFEB 介导的 ALP 增强似乎对神经元中的 Aβ 产生具有双重但相反的影响。