Dystrophin Deficiency Leads to Genomic Instability in Human Pluripotent Stem Cells via NO Synthase-Induced Oxidative Stress
Dystrophin Deficiency Leads to Genomic Instability in Human Pluripotent Stem Cells via NO Synthase-Induced Oxidative Stress
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DOI：
10.3390/cells8010053
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发表时间：
2019-01-01
期刊：
影响因子：
6
通讯作者：
Rotrekl, Vladimir
中科院分区：
文献类型：
--
作者：
Jelinkova, Sarka;Fojtik, Petr;Rotrekl, Vladimir
最近的数据显示，肌细胞祖细胞参与疾病的病理往往导致DMD患者的死亡。DMD中干细胞损伤的分子机制尚未描述。我们通过重编程来自两名DMD患者的细胞，以及通过CRISPR/Cas9将抗肌萎缩蛋白突变引入人胚胎干细胞，创建了抗肌萎缩蛋白缺陷型人多能干细胞（hPSC）系。虽然肌营养不良蛋白在健康hPSC中表达，但其在DMD hPSC系中的缺陷通过一氧化氮合酶的所有三种同种型的失调活性诱导活性氧类（ROS）的释放（进一步见下文）。as，NOS）。NOS诱导的ROS释放导致DMD hPSC的DNA损伤和基因组不稳定性。我们能够通过抑制NOS活性将ROS释放以及DNA损伤降低到野生型hPSC的水平。
Recent data on Duchenne muscular dystrophy (DMD) show myocyte progenitor's involvement in the disease pathology often leading to the DMD patient's death. The molecular mechanism underlying stem cell impairment in DMD has not been described. We created dystrophin-deficient human pluripotent stem cell (hPSC) lines by reprogramming cells from two DMD patients, and also by introducing dystrophin mutation into human embryonic stem cells via CRISPR/Cas9. While dystrophin is expressed in healthy hPSC, its deficiency in DMD hPSC lines induces the release of reactive oxygen species (ROS) through dysregulated activity of all three isoforms of nitric oxide synthase (further abrev. as, NOS). NOS-induced ROS release leads to DNA damage and genomic instability in DMD hPSC. We were able to reduce both the ROS release as well as DNA damage to the level of wild-type hPSC by inhibiting NOS activity.