Adding a temporal dimension to the study of Friedreich's ataxia: the effect of frataxin overexpression in a human cell model.
Adding a temporal dimension to the study of Friedreich's ataxia: the effect of frataxin overexpression in a human cell model.
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DOI：
10.1242/dmm.032706
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发表时间：
2018-06-25
影响因子：
4.3
通讯作者：
Pastore A
中科院分区：
文献类型：
--
作者：
Vannocci T;Notario Manzano R;Beccalli O;Bettegazzi B;Grohovaz F;Cinque G;de Riso A;Quaroni L;Codazzi F;Pastore A
神经退行性疾病弗里德赖希共济失调是由低于正常水平的共济失调蛋白引起的，共济失调蛋白是一种参与铁硫（Fe-S）簇生物发生的重要蛋白质。设计治疗这种疾病的策略的一个重要步骤是了解通过基因治疗增加共济失调蛋白水平是否只是有益或有害的，因为以前的研究，主要是基于动物模型，报告了相互矛盾的结果。在这里，我们利用了一种诱导型模型，我们使用CRISPR/Cas9方法开发了该模型，以研究frataxin过表达在人类细胞中的影响，并监测过表达后系统如何恢复。使用新的工具，从高通量显微镜到细胞内红外，我们证明了frataxin基因的过度表达会影响细胞代谢。它还导致氧化应激和不稳定的铁池水平显着增加。这些细胞变化与该基因部分沉默时观察到的变化相似，如Friedreich共济失调患者中发生的情况。我们的数据表明，共济失调蛋白的水平必须受到严格的调节和微调，任何不平衡都会导致氧化应激和毒性。总结：我们已经在诱导型细胞模型中探索了共济失调蛋白基因过表达的影响。我们的数据表明，必须严格调节共济失调蛋白的水平，因为任何不平衡都会导致氧化应激和毒性。
The neurodegenerative disease Friedreich's ataxia is caused by lower than normal levels of frataxin, an important protein involved in iron–sulfur (Fe-S) cluster biogenesis. An important step in designing strategies to treat this disease is to understand whether increasing the frataxin levels by gene therapy would simply be beneficial or detrimental, because previous studies, mostly based on animal models, have reported conflicting results. Here, we have exploited an inducible model, which we developed using the CRISPR/Cas9 methodology, to study the effects of frataxin overexpression in human cells and monitor how the system recovers after overexpression. Using new tools, which range from high-throughput microscopy to in cell infrared, we prove that overexpression of the frataxin gene affects the cellular metabolism. It also leads to a significant increase of oxidative stress and labile iron pool levels. These cellular alterations are similar to those observed when the gene is partly silenced, as occurs in Friedreich's ataxia patients. Our data suggest that the levels of frataxin must be tightly regulated and fine-tuned, with any imbalance leading to oxidative stress and toxicity. Summary: We have probed the effect of overexpression of the frataxin gene in an inducible cellular model. Our data indicate that the levels of frataxin must be tightly regulated, because any imbalance leads to oxidative stress and toxicity.
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DOI：
10.1038/mtm.2014.44
发表时间：
2014
期刊：
Molecular therapy. Methods & clinical development
影响因子：
--
作者：
通讯作者：
--
DOI：
10.1073/pnas.94.14.7452
发表时间：
1997-07-08
影响因子：
11.1
作者：
Cossee, M;Schmitt, M;Koenig, M
通讯作者：
Koenig, M
影响因子：
82.9
作者：
Perdomini, Morgane;Belbellaa, Brahim;Puccio, Helene
通讯作者：
Puccio, Helene
影响因子：
16.6
作者：
Prischi, Filippo;Konarev, Petr V.;Iannuzzi, Clara;Pastore, Chiara;Adinolfi, Salvatore;Martin, Stephen R.;Svergun, Dmitri I.;Pastore, Annalisa
通讯作者：
Pastore, Annalisa
影响因子：
3.7
作者：
Navarro JA;Llorens JV;Soriano S;Botella JA;Schneuwly S;Martínez-Sebastián MJ;Moltó MD
通讯作者：
Moltó MD