Defective lysosomal acidification contributes to TNFR1 mediated neuronal necroptosis in Alzheimer's disease

BackgroundTumor necrosis factor (TNF) receptor 1 (TNFR1) signaling mediates neuronal necroptosis in Alzheimers disease (AD). Interaction of TNFR1 signaling axis with autolysosomal pathway and the accumulation of necrosome molecules in impaired lysosomes have been shown to lead to necroptotic neuronal death. This has been attributed to the terminal failure of the autophagic process, primarily due to lysosomal degradation dysfunction. Being the final and determining step of the autolysosomal pathway, lysosomes with sufficient acidification as maintained by functional vacuolar (H+)-ATPase (V-ATPase) are required to achieve complete autophagic degradation of toxic cellular components. Here, we aim to investigate the role of defective lysosomal acidification in mediating TNFR1 induced neuronal necroptosis in AD. MethodsNeuropathological analysis of human post-mortem AD brains was performed to examine the correlation between TNFR1 induced neuronal necroptosis and autolysosomal dysfunction. Specifically, we probed for the level of V-ATPase subunits in AD brains to determine the extent of lysosomal acidification and function. Cell-based assays were conducted to understand the effect of TNFR1 activation in driving lysosomal acidification defect, autophagic impairment, mitochondrial dysfunction, and neuronal death in SH-SY5Y neuroblastoma cells. Furthermore, we applied lysosome-acidifying nanoparticles (AcNPs) to determine whether restoration of lysosomal acidification can rescue neuronal necroptosis in both TNF-treated SH-SY5Y cells and APPNL-G-F knock-in mouse model of AD. ResultsWe revealed that TNFR1 activated neuronal necroptosis correlates with autolysosomal dysfunction as characterized by downregulation of V-ATPase subunits and accumulation of autophagy receptor p62 in human AD brains. In cell culture, we showed for the first time that lysosomal acidification is only impaired in cells treated with TNF and not with other cytokines, contributing to inhibition of autophagic degradation in SH-SY5Y cells. We also illustrated that there is defective mitochondrial turnover, together with reduced mitochondrial functions and elevated reactive oxygen species, leading to neuronal death in SH-SY5Y cells. Importantly, we demonstrated that AcNPs restore lysosomal acidification, autophagic activity, and mitochondrial function, as well as rescue neuronal necroptosis in both TNF-treated SH-SY5Y cells and APPNL- G-F mice. ConclusionsDefective lysosomal acidification plays a key role in TNFR1 mediated neuronal necroptosis. This opens avenues for new therapeutic strategies to target lysosomal acidification dysfunction in AD.