In vitro supplementation with deoxynucleoside monophosphates rescues mitochondrial DNA depletion
In vitro supplementation with deoxynucleoside monophosphates rescues mitochondrial DNA depletion
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DOI：
10.1016/j.ymgme.2012.04.022
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发表时间：
2012-09-01
影响因子：
3.8
通讯作者：
Horvath, Rita
中科院分区：
文献类型：
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作者：
Bulst, Stefanie;Holinski-Feder, Elke;Horvath, Rita
线粒体DNA耗竭综合征是一组遗传异质性的通常严重的疾病，其特征在于细胞线粒体DNA含量减少。线粒体DNA耗竭综合征的潜在治疗策略的研究将依赖于良好的模型系统。我们以前曾建议，肌管可能是最佳的模型系统，这样的研究。在这里，我们首先验证了这种技术在不同范围的线粒体DNA耗竭综合征患者的细胞，显示在细胞系的遗传和表型不同的patients. Second，我们开发了一种假定的治疗方法，使用不同的组合脱氧核苷单磷酸在不同类型的线粒体DNA耗竭综合征，在许多情况下，显示接近正常化的线粒体DNA含量。此外，我们使用核苷类逆转录酶抑制剂在体外精确地滴定mtDNA消耗。以这种方式，我们可以揭示线粒体耗竭综合征细胞系中的生理缺陷，这也可以通过补充脱氧核苷一磷酸来改善。最后，我们将该模型扩展到研究通过MyoD转染的肌源性转分化后的成纤维细胞，其类似于原代肌管也显示脱氧核苷一磷酸反应性线粒体！本研究的结果提示，根据线粒体DNA缺失综合征患者的原发基因缺陷和分子机制，采用不同的脱氧核苷单磷酸联合用药，可能是一种治疗线粒体DNA缺失综合征的可行方法，值得临床进一步研究。(c)2012 Elsevier Inc. All rights reserved.
Mitochondrial DNA depletion syndromes are a genetically heterogeneous group of often severe diseases, characterized by reduced cellular mitochondrial DNA content. Investigation of potential therapeutic strategies for mitochondrial DNA depletion syndromes will be dependent on good model systems. We have previously suggested that myotubes may be the optimal model system for such studies. Here we firstly validate this technique in a diverse range of cells of patients with mitochondrial DNA depletion syndromes, showing contrasting effects in cell lines from genetically and phenotypically differing patients.Secondly, we developed a putative therapeutic approach using variable combinations of deoxynucleoside monophosphates in different types of mitochondrial DNA depletion syndromes, showing near normalization of mitochondrial DNA content in many cases. Furthermore, we used nucleoside reverse transcriptase inhibitors to precisely titrate mtDNA depletion in vitro. In this manner we can unmask a physiological defect in mitochondrial depletion syndrome cell lines which is also ameliorated by deoxynucleoside monophosphate supplementation. Finally, we have extended this model to study fibroblasts after myogenic transdifferentiation by MyoD transfection, which similar to primary myotubes also showed deoxynucleoside monophosphate responsive mitochondria! DNA depletion in vitro, thus providing a more convenient method for deriving future models of mitochondrial DNA depletion.Our results suggest that using different combinations of deoxynucleoside monophosphates depending on the primary gene defect and molecular mechanism may be a possible therapeutic approach for many patients with mitochondrial DNA depletion syndromes and is worthy of further clinical investigation. (c) 2012 Elsevier Inc. All rights reserved.