Department of Regenerative Medicine and Cell Biology, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC 29425, USA.
Department of Drug Discovery and Biomedical Sciences, Medical University of South Carolina, 173 Ashley Avenue, Charleston, SC 29425, USA.
Cell Rep. 2018 Nov 6;25(6):1469-1484.e5. doi: 10.1016/j.celrep.2018.10.036.
Patients with mtDNA depletion syndrome 3 (MTDPS3) often die as children from liver failure caused by severe reduction in mtDNA content. The identification of treatments has been impeded by an inability to culture and manipulate MTDPS3 primary hepatocytes. Here we generated DGUOK-deficient hepatocyte-like cells using induced pluripotent stem cells (iPSCs) and used them to identify drugs that could improve mitochondrial ATP production and mitochondrial function. Nicotinamide adenine dinucleotide (NAD) was found to improve mitochondrial function in DGUOK-deficient hepatocyte-like cells by activating the peroxisome proliferator-activated receptor gamma coactivator 1-alpha (PGC1α). NAD treatment also improved ATP production in MTDPS3-null rats and in hepatocyte-like cells that were deficient in ribonucleoside-diphosphate reductase subunit M2B (RRM2B), suggesting that it could be broadly effective. Our studies reveal that DGUOK-deficient iPSC-derived hepatocytes recapitulate the pathophysiology of MTDPS3 in culture and can be used to identify therapeutics for mtDNA depletion syndromes.
患有线粒体 DNA 耗竭综合征 3 型（MTDPS3）的患者通常会因 mtDNA 含量严重减少导致肝功能衰竭而在儿童时期死亡。由于无法培养和操纵 MTDPS3 原代肝细胞，因此难以确定治疗方法。在这里，我们使用诱导多能干细胞（iPSCs）生成了 DGUOK 缺陷型肝样细胞，并利用它们来鉴定可改善线粒体 ATP 生成和线粒体功能的药物。发现烟酰胺腺嘌呤二核苷酸（NAD）通过激活过氧化物酶体增殖物激活受体 γ 共激活因子 1-α（PGC1α）可改善 DGUOK 缺陷型肝样细胞中的线粒体功能。NAD 处理还改善了 MTDPS3 缺陷型大鼠和缺乏核昔二磷酸还原酶亚基 M2B（RRM2B）的肝样细胞中的 ATP 生成，这表明 NAD 可能具有广泛的治疗效果。我们的研究表明，由 DGUOK 缺陷型 iPSC 衍生的肝细胞在培养中再现了 MTDPS3 的病理生理学，可用于鉴定 mtDNA 耗竭综合征的治疗方法。
Zotero 插件