Metabolic rescue ameliorates mitochondrial encephalo-cardiomyopathy in murine and human iPSC models of Leigh syndrome.
Metabolic rescue ameliorates mitochondrial encephalo-cardiomyopathy in murine and human iPSC models of Leigh syndrome.
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代谢挽救改善Leigh综合征小鼠和人iPSC模型中的线粒体脑心肌病。
DOI：
10.1002/ctm2.954
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发表时间：
2022-07
影响因子：
10.6
通讯作者：
中科院分区：
文献类型：
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复合物 I 亚基 Ndufs4 缺失的小鼠会出现类似于 Leigh 综合征 (LS) 的线粒体脑肌病。 LS 心脑肌病的代谢紊乱和潜在机制仍不完全清楚。我们对缺乏 Ndufs4 的小鼠进行了超声心动图、电生理学、共聚焦显微镜、代谢和分子/形态测量分析。 HEK293 细胞、人 iPS 细胞衍生的心肌细胞和神经元用于确定线粒体复合物 I 缺陷的机制作用。 LS 小鼠出现严重的心律失常和舒张功能障碍。 Ndufs4 缺失的人诱导多能干细胞衍生心肌细胞 (iPS-CM) 再现了 LS 心肌病。从机制上讲，我们证明了复合物 I 缺乏、细胞内（烟酰胺腺嘌呤二核苷酸）NAD+/NADH 减少和缓慢性心律失常之间的直接联系，这是由心脏钠通道 NaV1.5（特别是在 K1479 位点）的过度乙酰化介导的。 LS 小鼠小脑和中脑区域的神经元凋亡与 p53 的过度乙酰化和小胶质细胞的激活有关。靶向代谢组学揭示了几种氨基酸和柠檬酸循环中间体的增加，可能是由于 NAD+ 依赖性脱氢酶的损伤，以及还原型谷胱甘肽 (GSH) 的大幅减少。通过补充烟酰胺核苷 (NR) 进行的代谢拯救可增加细胞内 NAD+/ NADH，恢复代谢紊乱，通过 NAD+ 依赖性 Sirtuin 脱乙酰酶逆转蛋白质过度乙酰化，并改善心肌病表型，同时改善 NaV1.5 电流和通过 Ca2+ 瞬变测量的 SERCA2a 功能。 NR 还减弱了 LS 脑和 Ndufs4 缺失的人 iPS 衍生神经元中的神经元凋亡和小胶质细胞活化。我们的研究揭示了在小鼠和人类诱导多能干细胞 (iPSC) LS 模型中观察到的心脏缓慢性心律失常、舒张功能障碍和神经元凋亡的直接机制解释。 Leigh 综合征 (LS) 是一种严重的线粒体脑肌病，可能累及心脏。其代谢紊乱的特点是 NAD+ 缺乏以及几种 NAD+ 依赖性代谢酶和 Sirtuins 的损伤。 Nicotinamide Riboside 可恢复 Sirt1 功能，逆转多种关键蛋白的过度乙酰化，包括心肌细胞中的 Nav1.5 和 SERCA2 以及大脑中的 p53，从而改善心动过缓、神经元丢失和脑病。
Mice with deletion of complex I subunit Ndufs4 develop mitochondrial encephalomyopathy resembling Leigh syndrome (LS). The metabolic derangement and underlying mechanisms of cardio‐encephalomyopathy in LS remains incompletely understood. We performed echocardiography, electrophysiology, confocal microscopy, metabolic and molecular/morphometric analysis of the mice lacking Ndufs4. HEK293 cells, human iPS cells‐derived cardiomyocytes and neurons were used to determine the mechanistic role of mitochondrial complex I deficiency. LS mice develop severe cardiac bradyarrhythmia and diastolic dysfunction. Human‐induced pluripotent stem cell‐derived cardiomyocytes (iPS‐CMs) with Ndufs4 deletion recapitulate LS cardiomyopathy. Mechanistically, we demonstrate a direct link between complex I deficiency, decreased intracellular (nicotinamide adenine dinucleotide) NAD+/NADH and bradyarrhythmia, mediated by hyperacetylation of the cardiac sodium channel NaV1.5, particularly at K1479 site. Neuronal apoptosis in the cerebellar and midbrain regions in LS mice was associated with hyperacetylation of p53 and activation of microglia. Targeted metabolomics revealed increases in several amino acids and citric acid cycle intermediates, likely due to impairment of NAD+‐dependent dehydrogenases, and a substantial decrease in reduced Glutathione (GSH). Metabolic rescue by nicotinamide riboside (NR) supplementation increased intracellular NAD+/ NADH, restored metabolic derangement, reversed protein hyperacetylation through NAD+‐dependent Sirtuin deacetylase, and ameliorated cardiomyopathic phenotypes, concomitant with improvement of NaV1.5 current and SERCA2a function measured by Ca2+‐transients. NR also attenuated neuronal apoptosis and microglial activation in the LS brain and human iPS‐derived neurons with Ndufs4 deletion. Our study reveals direct mechanistic explanations of the observed cardiac bradyarrhythmia, diastolic dysfunction and neuronal apoptosis in mouse and human induced pluripotent stem cells (iPSC) models of LS. Leigh syndrome (LS) is a severe mitochondrial encephalomyopathy that may involve heart. Its metabolic derangement is characterized by NAD+ deficiency and impairment of several NAD+‐dependent metabolic enzymes and Sirtuins. Nicotinamide Riboside restores Sirt1 function, reverses hyperacetylation of several crucial proteins, including Nav1.5 and SERCA2 in cardiomyocytes, and p53 in the brain, leading to amelioration of bradycardia, neuronal loss and encephalopathy.
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DOI：
10.1073/pnas.1113987109
发表时间：
2012-04-17
影响因子：
11.1
作者：
Ke, Bi-Xia;Pepe, Salvatore;Thorburn, David R.
通讯作者：
Thorburn, David R.
影响因子：
20.1
作者：
Davidson, Michael T.;Grimsrud, Paul A.;Lai, Ling;Draper, James A.;Fisher-Wellman, Kelsey H.;Narowski, Tara M.;Abraham, Dennis M.;Koves, Timothy R.;Kelly, Daniel P.;Muoio, Deborah M.
通讯作者：
Muoio, Deborah M.
影响因子：
3.7
作者：
Chouchani ET;Methner C;Buonincontri G;Hu CH;Logan A;Sawiak SJ;Murphy MP;Krieg T
通讯作者：
Krieg T
影响因子：
3.8
作者：
Johnson, Simon C.;Kayser, Ernst-Bernhard;Morgan, Philip G.
通讯作者：
Morgan, Philip G.
影响因子：
64.5
作者：
Gu, W;Roeder, RG
通讯作者：
Roeder, RG