EpiPro, a Novel, Synthetic, Activity-Regulated Promoter That Targets Hyperactive Neurons in Epilepsy for Gene Therapy Applications.
EpiPro, a Novel, Synthetic, Activity-Regulated Promoter That Targets Hyperactive Neurons in Epilepsy for Gene Therapy Applications.
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DOI：
10.3390/ijms241914467
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发表时间：
2023-09-23
影响因子：
5.6
通讯作者：
Perez-Reyes E
中科院分区：
文献类型：
--
作者：
Burke CT;Vitko I;Straub J;Nylund EO;Gawda A;Blair K;Sullivan KA;Ergun L;Ottolini M;Patel MK;Perez-Reyes E
癫痫发生的特征在于神经元放电的内在变化，导致神经元过度活跃和随后产生癫痫发作活动。这些改变伴随着基因转录网络的变化，首先是早期即时基因的激活，然后是参与记忆的基因的长期激活。我们的目标是设计一个启动子，该启动子包含慢性癫痫中上调的活性依赖性转录因子（EpiPro）的结合位点，并在多种啮齿动物癫痫模型中验证。首先，我们评估了EpiPro的活性依赖性：最初的电生理学研究发现，与未标记的神经元相比，EpiPro驱动的GFP表达与放电率增加有关，EpiPro驱动的GFP表达的评估显示，癫痫持续状态后GFP表达增加约150倍。在此之后，我们比较了EpiPro驱动的GFP表达在两种啮齿动物癫痫模型，大鼠锂/毛果芸香碱和小鼠电点燃。在慢性癫痫的啮齿类动物中，GFP表达在大多数神经元中增加，但特别是在齿状颗粒细胞中，提供了体内证据支持边缘癫痫发生的“齿状门崩溃”假说。最后，我们评估了EpiPro激活的时间过程，发现它在癫痫发作后迅速诱导，在数周后失活，证实了EpiPro作为癫痫基因治疗驱动剂的潜在效用。
Epileptogenesis is characterized by intrinsic changes in neuronal firing, resulting in hyperactive neurons and the subsequent generation of seizure activity. These alterations are accompanied by changes in gene transcription networks, first with the activation of early-immediate genes and later with the long-term activation of genes involved in memory. Our objective was to engineer a promoter containing binding sites for activity-dependent transcription factors upregulated in chronic epilepsy (EpiPro) and validate it in multiple rodent models of epilepsy. First, we assessed the activity dependence of EpiPro: initial electrophysiology studies found that EpiPro-driven GFP expression was associated with increased firing rates when compared with unlabeled neurons, and the assessment of EpiPro-driven GFP expression revealed that GFP expression was increased ~150× after status epilepticus. Following this, we compared EpiPro-driven GFP expression in two rodent models of epilepsy, rat lithium/pilocarpine and mouse electrical kindling. In rodents with chronic epilepsy, GFP expression was increased in most neurons, but particularly in dentate granule cells, providing in vivo evidence to support the “breakdown of the dentate gate” hypothesis of limbic epileptogenesis. Finally, we assessed the time course of EpiPro activation and found that it was rapidly induced after seizures, with inactivation following over weeks, confirming EpiPro’s potential utility as a gene therapy driver for epilepsy.
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影响因子：
3.7
作者：
Lim JA;Moon J;Kim TJ;Jun JS;Park B;Byun JI;Sunwoo JS;Park KI;Lee ST;Jung KH;Jung KY;Kim M;Jeon D;Chu K;Lee SK
通讯作者：
Lee SK
影响因子：
12.4
作者：
Manfredsson, Fredric P.;Burger, Corinna;Mandel, Ronald J.
通讯作者：
Mandel, Ronald J.
影响因子：
5.1
作者：
通讯作者：
--
影响因子：
12.4
作者：
Klein, Ronald L.;Dayton, Robert D.;Henning, Phillip P.
通讯作者：
Henning, Phillip P.
DOI：
10.1073/pnas.0806518106
发表时间：
2009-01-06
影响因子：
11.1
作者：
Kawashima, Takashi;Okuno, Hiroyuki;Bito, Haruhiko
通讯作者：
Bito, Haruhiko