Wilson Cathal, Giaquinto Laura, Santoro Michele, Di Tullio Giuseppe, Morra Valentina, Kukulski Wanda, Venditti Rossella, Navone Francesca, Borgese Nica, De Matteis Maria Antonietta
Telethon Institute of Genetics and Medicine, TIGEM, Pozzuoli, Italy
Department of Molecular Medicine and Medical Biotechnology, University of Naples Federico II, Naples, Italy.
Life Sci Alliance. 2025 Jan 27;8(4). doi: 10.26508/lsa.202402907. Print 2025 Apr.
Protein aggregates in motoneurons, a pathological hallmark of amyotrophic lateral sclerosis, have been suggested to play a key pathogenetic role. ALS8, characterized by ER-associated inclusions, is caused by a heterozygous mutation in VAPB, which acts at multiple membrane contact sites between the ER and almost all other organelles. The link between protein aggregation and cellular dysfunction is unclear. A yeast model, expressing human mutant and WT-VAPB under the control of the orthologous yeast promoter in haploid and diploid cells, was developed to mimic the disease situation. Inclusion formation was found to be a developmentally regulated process linked to mitochondrial damage that could be attenuated by reducing ER-mitochondrial contacts. The co-expression of the WT protein retarded P56S-VAPB inclusion formation. Importantly, we validated these results in mammalian motoneuron cells. Our findings indicate that (age-related) damage to mitochondria influences the propensity of the mutant VAPB to form aggregates via ER-mitochondrial contacts, initiating a series of events leading to disease progression.
运动神经元中的蛋白质聚集体是肌萎缩侧索硬化症的一个病理标志,被认为起着关键的致病作用。以内质网相关包涵体为特征的ALS8是由VAPB中的杂合突变引起的,VAPB在内质网与几乎所有其他细胞器之间的多个膜接触位点发挥作用。蛋白质聚集与细胞功能障碍之间的联系尚不清楚。我们开发了一种酵母模型,在单倍体和二倍体细胞中,在直系同源酵母启动子的控制下表达人类突变型和野生型VAPB,以模拟疾病情况。发现包涵体形成是一个与线粒体损伤相关的发育调控过程,通过减少内质网-线粒体接触可以减弱这种损伤。野生型蛋白的共表达延缓了P56S-VAPB包涵体的形成。重要的是,我们在哺乳动物运动神经元细胞中验证了这些结果。我们的研究结果表明,(与年龄相关的)线粒体损伤通过内质网-线粒体接触影响突变型VAPB形成聚集体的倾向,引发一系列导致疾病进展的事件。