Shaw K T, Utsuki T, Rogers J, Yu Q S, Sambamurti K, Brossi A, Ge Y W, Lahiri D K, Greig N H
Drug Design and Development, Laboratory of Neurosciences, National Institute on Aging, Baltimore, MD 21224, USA.
Proc Natl Acad Sci U S A. 2001 Jun 19;98(13):7605-10. doi: 10.1073/pnas.131152998. Epub 2001 Jun 12.
The reduction in levels of the potentially toxic amyloid-beta peptide (Abeta) has emerged as one of the most important therapeutic goals in Alzheimer's disease. Key targets for this goal are factors that affect the expression and processing of the Abeta precursor protein (betaAPP). Earlier reports from our laboratory have shown that a novel cholinesterase inhibitor, phenserine, reduces betaAPP levels in vivo. Herein, we studied the mechanism of phenserine's actions to define the regulatory elements in betaAPP processing. Phenserine treatment resulted in decreased secretion of soluble betaAPP and Abeta into the conditioned media of human neuroblastoma cells without cellular toxicity. The regulation of betaAPP protein expression by phenserine was posttranscriptional as it suppressed betaAPP protein expression without altering betaAPP mRNA levels. However, phenserine's action was neither mediated through classical receptor signaling pathways, involving extracellular signal-regulated kinase or phosphatidylinositol 3-kinase activation, nor was it associated with the anticholinesterase activity of the drug. Furthermore, phenserine reduced expression of a chloramphenicol acetyltransferase reporter fused to the 5'-mRNA leader sequence of betaAPP without altering expression of a control chloramphenicol acetyltransferase reporter. These studies suggest that phenserine reduces Abeta levels by regulating betaAPP translation via the recently described iron regulatory element in the 5'-untranslated region of betaAPP mRNA, which has been shown previously to be up-regulated in the presence of interleukin-1. This study identifies an approach for the regulation of betaAPP expression that can result in a substantial reduction in the level of Abeta.
降低潜在毒性的β淀粉样肽(Aβ)水平已成为阿尔茨海默病最重要的治疗目标之一。实现这一目标的关键靶点是影响Aβ前体蛋白(βAPP)表达和加工的因素。我们实验室早期的报告表明,一种新型胆碱酯酶抑制剂苯丝氨酸可在体内降低βAPP水平。在此,我们研究了苯丝氨酸的作用机制,以确定βAPP加工过程中的调控元件。苯丝氨酸处理导致可溶性βAPP和Aβ分泌到人类神经母细胞瘤细胞的条件培养基中减少,且无细胞毒性。苯丝氨酸对βAPP蛋白表达的调节是转录后调节,因为它抑制βAPP蛋白表达而不改变βAPP mRNA水平。然而,苯丝氨酸的作用既不是通过涉及细胞外信号调节激酶或磷脂酰肌醇3激酶激活的经典受体信号通路介导的,也与该药物的抗胆碱酯酶活性无关。此外,苯丝氨酸降低了与βAPP的5'-mRNA前导序列融合的氯霉素乙酰转移酶报告基因的表达,而不改变对照氯霉素乙酰转移酶报告基因的表达。这些研究表明,苯丝氨酸通过βAPP mRNA 5'-非翻译区中最近描述的铁反应元件调节βAPP翻译来降低Aβ水平,先前已证明该元件在白细胞介素-1存在时会上调。本研究确定了一种调节βAPP表达的方法,该方法可导致Aβ水平大幅降低。