Institute of Genetics and Biotechnology, Faculty of Biology, University of Warsaw, 5a Pawińskiego Str., 02-106, Warsaw, Poland.
Faculty of Physics, University of Warsaw, 5 Pasteur Str., 02-093, Warsaw, Poland.
J Appl Genet. 2020 May;61(2):195-203. doi: 10.1007/s13353-020-00550-y. Epub 2020 Mar 10.
Leber's hereditary optic neuropathy (LHON) is one of the most common mitochondrial diseases caused by point mutations in mitochondrial DNA (mtDNA). The majority of diagnosed LHON cases are caused by a point mutation at position 11,778 in the mitochondrial genome. LHON mainly affects young men in their 20s and 30s with usually poor visual prognosis. It remains unexplained why men are more likely to develop the disease and why only retinal ganglion cells are affected. In this study, a cell model was used for the first time to investigate the influence of testosterone on the cell death mechanism apoptosis and on an autophagy/mitophagy. Cells with m.11778G > A were found to be significantly more susceptible to nucleosome formation and effector caspase activation that serve as hallmarks of apoptotic cell death. Cells having this mutation expressed higher levels of mitophagic receptors BNIP3 and BNIP3L/Nix in a medium with testosterone. Moreover, cells having the mutation exhibited greater mitochondrial mass, which suggests these cells have a decreased cell survival. The observed decrease in cell survival was supported by the observed increase in apoptotic cell death. Autophagy was analyzed after inhibition with Bafilomycin A1 (Baf A1). The results indicate impairment in autophagy in LHON cells due to lower autophagic flux supported by observed lower levels of autophagosome marker LC3-II. The observed impaired lower autophagic flux in mutant cells correlated with increased levels of BNIP3 and BNIP3L/Nix in mutant cells.
Leber 遗传性视神经病变(LHON)是由线粒体 DNA(mtDNA)点突变引起的最常见的线粒体疾病之一。大多数诊断为 LHON 的病例是由线粒体基因组中位置 11,778 的点突变引起的。LHON 主要影响 20 多岁和 30 多岁的年轻男性,通常视力预后较差。目前仍不清楚为什么男性更容易患这种疾病,以及为什么只有视网膜神经节细胞受到影响。在这项研究中,首次使用细胞模型研究了睾丸酮对细胞死亡机制凋亡和自噬/线粒体自噬的影响。研究发现,携带 m.11778G > A 的细胞更容易形成核小体并激活效应半胱天冬酶,这是细胞凋亡的特征。在含有睾丸酮的培养基中,携带这种突变的细胞表达更高水平的线粒体自噬受体 BNIP3 和 BNIP3L/Nix。此外,携带这种突变的细胞表现出更大的线粒体质量,这表明这些细胞的细胞存活率降低。观察到的细胞存活率下降得到了凋亡细胞死亡增加的支持。在用 Bafilomycin A1(Baf A1)抑制后分析自噬。结果表明,由于观察到的自噬小体标记物 LC3-II 水平较低,LHON 细胞中的自噬受到损害,自噬通量降低。在突变细胞中观察到的受损的较低自噬通量与突变细胞中 BNIP3 和 BNIP3L/Nix 水平的增加相关。