Nonamyloidogenic processing of amyloid precursor protein (APP) by augmenting ADAM10 is a promising therapeutic strategy for Alzheimer’s disease (AD). Therefore identification of molecular pathways that regulate ADAM10 expression is crucial. Autophagy is strongly dysregulated in AD, and TFEB was recently shown to be a master regulator of autophagy-lysosome pathway (ALP). Here, we report that TFEB expression in HeLa cells increased ADAM10 mature form by 72% (p < 0.01, n = 4), while TFEB knockdown by CRISPR strategy reduced ADAM10 mature form by 36% (p < 0.05, n = 4). Autophagy inhibition by 3-methyladenine (3-MA), but not bafilomycin A1 (BAF1), reduced ADAM10 mature form by 49% (p < 0.05, n = 4) in the TFEB expressing HeLa cells. Autophagy activation by 3 h of starvation increased ADAM10 to 91% (p < 0.001, n = 6) relative to 51% (p < 0.01, n = 6) in the nutrient-fed cells. Further, siRNAs targeted against PPARα in HeLa cells decreased ADAM10 levels by 28% (p < 0.05, n = 6) relative to the cells treated with scrambled siRNAs. Further, incubation of EGFP-TFEB expressing HeLa cells with PPARα antagonist, but not PPARβ or PPARγ antagonists, prevented TFEB-induced increase in ADAM10 levels. Importantly, flag-TFEB expression in the brain also increased ADAM10 by 60% (p < 0.05, n = 3) in the cortical and 34% (p < 0.001, n = 3) in the hippocampal homogenates. ADAM10 activity also increased by 57% (p < 0.01, n = 3) in the HeLa cells. Finally, TFEB-induced ADAM10 potentiation led to increased secretion of sAPPα by 154% (p < 0.001, n = 3) in the cortex and 62% (p < 0.001, n = 3) in the hippocampus. Thus, TFEB expression enhances nonamyloidogenic processing of APP. In conclusion, TFEB expression induces ADAM10 in an autophagy-dependent manner through PPARα.

通过增强 ADAM10 对淀粉样前体蛋白 (APP) 进行非淀粉样蛋白加工是阿尔茨海默病 (AD) 的一种有前途的治疗策略。因此，鉴定调节 ADAM10 表达的分子途径至关重要。自噬在 AD 中严重失调，并且最近显示 TFEB 是自噬溶酶体途径 (ALP) 的主要调节因子。在这里，我们报告 HeLa 细胞中的 TFEB 表达使 ADAM10 成熟形式增加了 72%（p  < 0.01，n  = 4），而 CRISPR 策略的 TFEB 敲低使 ADAM10 成熟形式减少了 36%（p  < 0.05，n  = 4）。3-甲基腺嘌呤 (3-MA) 而不是巴弗洛霉素 A1 (BAF1) 的自噬抑制使 ADAM10 成熟形式减少了 49% ( p  < 0.05, n = 4) 在表达 TFEB 的 HeLa 细胞中。饥饿 3 小时后的自噬激活将 ADAM10 增加至 91%（p  < 0.001，n  = 6），而营养喂养细胞中的 ADAM10 为 51%（p  < 0.01，n  = 6）。此外，相对于用乱序 siRNA 处理的细胞，靶向 HeLa 细胞中 PPARα 的 siRNA 使 ADAM10 水平降低了 28%（p  < 0.05，n  = 6）。此外，将表达 EGFP-TFEB 的 HeLa 细胞与 PPARα 拮抗剂（而不是 PPARβ 或 PPARγ 拮抗剂）一起孵育可防止 TFEB 诱导的 ADAM10 水平升高。重要的是，大脑中 flag-TFEB 的表达也使皮质中的 ADAM10 增加了 60%（p  < 0.05，n  = 3）和 34%（p < 0.001, n  = 3) 在海马匀浆中。HeLa 细胞中的ADAM10 活性也增加了 57% ( p  < 0.01, n  = 3)。最后，TFEB 诱导的 ADAM10 增强导致皮质中 sAPPα 的分泌增加 154%（p  < 0.001，n  = 3）和海马中 62%（p  < 0.001，n  = 3）。因此，TFEB 表达增强了 APP 的非淀粉样蛋白加工。总之，TFEB 表达通过 PPARα 以自噬依赖性方式诱导 ADAM10。