Siano Giacomo, Caiazza Maria Claudia, Ollà Ivana, Varisco Martina, Madaro Giuseppe, Quercioli Valentina, Calvello Mariantonietta, Cattaneo Antonino, Di Primio Cristina
Laboratorio di Biologia (BIO@SNS), Scuola Normale Superiore, Pisa, Italy.
Neurotrophins and Neurodegenerative Diseases Laboratory, Rita Levi-Montalcini European Brain Research Institute, Rome, Italy.
Front Cell Neurosci. 2019 Aug 21;13:386. doi: 10.3389/fncel.2019.00386. eCollection 2019.
Formation of Tau aggregates is a common pathological feature of tauopathies and their accumulation directly correlates with cytotoxicity and neuronal degeneration. Great efforts have been made to understand Tau aggregation and to find therapeutics halting or reversing the process, however, progress has been slowed due to the lack of a suitable method for monitoring Tau aggregation. We developed a cell-based assay allowing to detect and quantify Tau aggregation in living cells. The system is based on the FRET biosensor CST able to monitor the molecular dynamic of Tau aggregation in different cellular conditions. We probed candidate compounds that could block Tau hyperphosphorylation. In particular, to foster the drug discovery process, we tested kinase inhibitors approved for the treatment of other diseases. We identified the ERK inhibitor PD-901 as a promising therapeutic molecule since it reduces and prevents Tau aggregation. This evidence establishes the CST cell-based aggregation assay as a reliable tool for drug discovery and suggests that PD-901 might be a promising compound to be tested for further preclinical studies on AD.
Tau蛋白聚集体的形成是tau蛋白病的常见病理特征，其积累与细胞毒性和神经元变性直接相关。人们为了解Tau蛋白聚集并寻找能够阻止或逆转这一过程的治疗方法付出了巨大努力，然而，由于缺乏监测Tau蛋白聚集的合适方法，进展一直较为缓慢。我们开发了一种基于细胞的检测方法，能够在活细胞中检测和量化Tau蛋白聚集。该系统基于FRET生物传感器CST，能够监测不同细胞条件下Tau蛋白聚集的分子动态。我们筛选了能够阻断Tau蛋白过度磷酸化的候选化合物。特别是，为了推动药物发现进程，我们测试了已被批准用于治疗其他疾病的激酶抑制剂。我们确定ERK抑制剂PD - 901是一种有前景的治疗分子，因为它能够减少并预防Tau蛋白聚集。这一证据确立了基于CST细胞的聚集检测方法作为药物发现的可靠工具，并表明PD - 901可能是一种有前景的化合物，有待在阿尔茨海默病的进一步临床前研究中进行测试。
