Yang Tien-Chun, Yarmishyn Aliaksandr A, Yang Yi-Ping, Lu Pin-Chen, Chou Shih-Jie, Wang Mong-Lien, Lin Tai-Chi, Hwang De-Kuang, Chou Yu-Bai, Chen Shih-Jen, Yu Wei-Kuang, Wang An-Guor, Hsu Chih-Chien, Chiou Shih-Hwa
Department of Medical Research, Taipei Veterans General Hospital, Taipei 11217, Taiwan.
School of Pharmaceutical Sciences, National Yang-Ming University, Taipei 11221, Taiwan.
Hum Mol Genet. 2020 Jun 3;29(9):1454-1464. doi: 10.1093/hmg/ddaa063.
The mutations in the genes encoding the subunits of complex I of the mitochondrial electron transport chain are the most common cause of Leber's hereditary optic neuropathy (LHON), a maternal hereditary disease characterized by retinal ganglion cell (RGC) degeneration. The characteristics of incomplete penetrance indicate that nuclear genetic and environmental factors also determine phenotypic expression of LHON. Therefore, further understanding of the role of mutant mitochondrial nicotinamide adenine dinucleotide dehydrogenase subunit proteins and nuclear genetic factors/environmental effects in the etiology of LHON is needed. In this study, we generated human-induced pluripotent stem cells (hiPSCs) from healthy control, unaffected LHON mutation carrier, and affected LHON patient. hiPSC-derived RGCs were used to study the differences between affected and unaffected carriers of mitochondrial DNA point mutation m.11778G > A in the MT-ND4 gene. We found that both mutated cell lines were characterized by increase in reactive oxygen species production, however, only affected cell line had increased levels of apoptotic cells. We found a significant increase in retrograde mitochondria and a decrease in stationary mitochondria in the affected RGC axons. In addition, the messenger RNA and protein levels of KIF5A in the LHON-affected RGCs were significantly reduced. Antioxidant N-acetyl-L-cysteine could restore the expression of KIF5A and the normal pattern of mitochondrial movement in the affected RGCs. To conclude, we found essential differences in the mutually dependent processes of oxidative stress, mitochondrial transport and apoptosis between two LHON-specific mutation carrier RGC cell lines, asymptomatic carrier and disease-affected, and identified KIF5A as a central modulator of these differences.
线粒体电子传递链复合体I亚基编码基因的突变是Leber遗传性视神经病变(LHON)最常见的病因,LHON是一种母系遗传性疾病,其特征为视网膜神经节细胞(RGC)退化。不完全外显的特征表明,核遗传和环境因素也决定了LHON的表型表达。因此,需要进一步了解突变的线粒体烟酰胺腺嘌呤二核苷酸脱氢酶亚基蛋白以及核遗传因素/环境效应在LHON病因学中的作用。在本研究中,我们从健康对照、未受影响的LHON突变携带者和受影响的LHON患者中生成了人诱导多能干细胞(hiPSC)。hiPSC衍生的RGC用于研究MT-ND4基因中线粒体DNA点突变m.11778G>A的受影响和未受影响携带者之间的差异。我们发现,两个突变细胞系的特征均为活性氧产生增加,然而,只有受影响的细胞系凋亡细胞水平升高。我们发现,受影响的RGC轴突中逆行线粒体显著增加,静止线粒体减少。此外,LHON受影响的RGC中KIF5A的信使核糖核酸和蛋白质水平显著降低。抗氧化剂N-乙酰-L-半胱氨酸可以恢复受影响的RGC中KIF5A的表达以及线粒体运动的正常模式。总之,我们发现了两种LHON特异性突变携带者RGC细胞系(无症状携带者和疾病患者)在氧化应激、线粒体运输和凋亡这些相互依赖过程中的本质差异,并确定KIF5A是这些差异的核心调节因子。