Biosciences Institute, Faculty of Medical Sciences, Newcastle University, Newcastle upon Tyne, NE4 5PL, UK.
Department of Neurology, Institute of Medicine, University of Tsukuba, Tsukuba, Ibaraki, 305-8575, Japan.
Cell Death Dis. 2024 May 31;15(5):382. doi: 10.1038/s41419-024-06770-y.
Impairment of autophagy leads to an accumulation of misfolded proteins and damaged organelles and has been implicated in plethora of human diseases. Loss of autophagy in actively respiring cells has also been shown to trigger metabolic collapse mediated by the depletion of nicotinamide adenine dinucleotide (NAD) pools, resulting in cell death. Here we found that the deficit in the autophagy-NAD axis underpins the loss of viability in cell models of a neurodegenerative lysosomal storage disorder, Niemann-Pick type C1 (NPC1) disease. Defective autophagic flux in NPC1 cells resulted in mitochondrial dysfunction due to impairment of mitophagy, leading to the depletion of both the reduced and oxidised forms of NAD as identified via metabolic profiling. Consequently, exhaustion of the NAD pools triggered mitochondrial depolarisation and apoptotic cell death. Our chemical screening identified two FDA-approved drugs, celecoxib and memantine, as autophagy activators which effectively restored autophagic flux, NAD levels, and cell viability of NPC1 cells. Of biomedical relevance, either pharmacological rescue of the autophagy deficiency or NAD precursor supplementation restored NAD levels and improved the viability of NPC1 patient fibroblasts and induced pluripotent stem cell (iPSC)-derived cortical neurons. Together, our findings identify the autophagy-NAD axis as a mechanism of cell death and a target for therapeutic interventions in NPC1 disease, with a potential relevance to other neurodegenerative disorders.
自噬功能的损伤会导致错误折叠的蛋白质和受损的细胞器积累,并与许多人类疾病有关。在活跃呼吸的细胞中,自噬的丧失也被证明会触发由烟酰胺腺嘌呤二核苷酸(NAD)池耗竭介导的代谢崩溃,导致细胞死亡。在这里,我们发现自噬-NAD 轴的缺陷是神经退行性溶酶体贮积症尼曼-匹克 C1(NPC1)病细胞模型中活力丧失的基础。NPC1 细胞中自噬通量的缺陷导致线粒体功能障碍,原因是自噬体的形成受损,从而导致 NAD 的还原和氧化形式都耗尽,这通过代谢谱分析得以确定。因此,NAD 池的枯竭引发线粒体去极化和凋亡性细胞死亡。我们的化学筛选确定了两种 FDA 批准的药物,塞来昔布和美金刚,作为自噬激活剂,可有效恢复 NPC1 细胞的自噬通量、NAD 水平和细胞活力。具有生物医学相关性的是,自噬缺陷的药理学挽救或 NAD 前体的补充恢复了 NAD 水平并提高了 NPC1 患者成纤维细胞和诱导多能干细胞(iPSC)衍生的皮质神经元的活力。总之,我们的研究结果确定了自噬-NAD 轴是 NPC1 疾病中细胞死亡的机制和治疗干预的靶点,对其他神经退行性疾病也具有潜在的相关性。