Sarkar Sovan, Perlstein Ethan O, Imarisio Sara, Pineau Sandra, Cordenier Axelle, Maglathlin Rebecca L, Webster John A, Lewis Timothy A, O'Kane Cahir J, Schreiber Stuart L, Rubinsztein David C
Department of Medical Genetics, University of Cambridge, Cambridge Institute for Medical Research, Addenbrooke's Hospital, Hills Road, Cambridge CB2 2XY, UK.
Nat Chem Biol. 2007 Jun;3(6):331-8. doi: 10.1038/nchembio883. Epub 2007 May 7.
The target of rapamycin proteins regulate various cellular processes including autophagy, which may play a protective role in certain neurodegenerative and infectious diseases. Here we show that a primary small-molecule screen in yeast yields novel small-molecule modulators of mammalian autophagy. We first identified new small-molecule enhancers (SMER) and inhibitors (SMIR) of the cytostatic effects of rapamycin in Saccharomyces cerevisiae. Three SMERs induced autophagy independently of rapamycin in mammalian cells, enhancing the clearance of autophagy substrates such as mutant huntingtin and A53T alpha-synuclein, which are associated with Huntington's disease and familial Parkinson's disease, respectively. These SMERs, which seem to act either independently or downstream of the target of rapamycin, attenuated mutant huntingtin-fragment toxicity in Huntington's disease cell and Drosophila melanogaster models, which suggests therapeutic potential. We also screened structural analogs of these SMERs and identified additional candidate drugs that enhanced autophagy substrate clearance. Thus, we have demonstrated proof of principle for a new approach for discovery of small-molecule modulators of mammalian autophagy.
雷帕霉素靶蛋白调节包括自噬在内的多种细胞过程,自噬在某些神经退行性疾病和感染性疾病中可能发挥保护作用。在此我们表明,在酵母中进行的一次小分子初筛产生了哺乳动物自噬的新型小分子调节剂。我们首先在酿酒酵母中鉴定出雷帕霉素细胞生长抑制作用的新小分子增强剂(SMER)和抑制剂(SMIR)。三种SMER在哺乳动物细胞中独立于雷帕霉素诱导自噬,增强了自噬底物如突变型亨廷顿蛋白和A53Tα-突触核蛋白的清除,它们分别与亨廷顿舞蹈病和家族性帕金森病相关。这些SMER似乎独立于雷帕霉素靶点或在其下游起作用,在亨廷顿舞蹈病细胞和果蝇模型中减弱了突变型亨廷顿蛋白片段的毒性,这表明其具有治疗潜力。我们还筛选了这些SMER的结构类似物,并鉴定出了其他增强自噬底物清除的候选药物。因此,我们已经证明了发现哺乳动物自噬小分子调节剂新方法的原理。