Tang Xin, Kim Julie, Zhou Li, Wengert Eric, Zhang Lei, Wu Zheng, Carromeu Cassiano, Muotri Alysson R, Marchetto Maria C N, Gage Fred H, Chen Gong
Department of Biology, Huck Institutes of Life Sciences, Pennsylvania State University, University Park, PA 16802;
Bucknell University, Lewisburg, PA 17837;
Proc Natl Acad Sci U S A. 2016 Jan 19;113(3):751-6. doi: 10.1073/pnas.1524013113. Epub 2016 Jan 5.
Rett syndrome is a severe form of autism spectrum disorder, mainly caused by mutations of a single gene methyl CpG binding protein 2 (MeCP2) on the X chromosome. Patients with Rett syndrome exhibit a period of normal development followed by regression of brain function and the emergence of autistic behaviors. However, the mechanism behind the delayed onset of symptoms is largely unknown. Here we demonstrate that neuron-specific K(+)-Cl(-) cotransporter2 (KCC2) is a critical downstream gene target of MeCP2. We found that human neurons differentiated from induced pluripotent stem cells from patients with Rett syndrome showed a significant deficit in KCC2 expression and consequently a delayed GABA functional switch from excitation to inhibition. Interestingly, overexpression of KCC2 in MeCP2-deficient neurons rescued GABA functional deficits, suggesting an important role of KCC2 in Rett syndrome. We further identified that RE1-silencing transcriptional factor, REST, a neuronal gene repressor, mediates the MeCP2 regulation of KCC2. Because KCC2 is a slow onset molecule with expression level reaching maximum later in development, the functional deficit of KCC2 may offer an explanation for the delayed onset of Rett symptoms. Our studies suggest that restoring KCC2 function in Rett neurons may lead to a potential treatment for Rett syndrome.
瑞特综合征是一种严重的自闭症谱系障碍,主要由X染色体上的单个基因甲基CpG结合蛋白2(MeCP2)突变引起。瑞特综合征患者在经历一段正常发育时期后,会出现脑功能衰退和自闭症行为。然而,症状延迟出现背后的机制在很大程度上尚不清楚。在此,我们证明神经元特异性钾氯共转运体2(KCC2)是MeCP2的关键下游基因靶点。我们发现,从瑞特综合征患者的诱导多能干细胞分化而来的人类神经元,其KCC2表达存在显著缺陷,因此γ-氨基丁酸(GABA)从兴奋到抑制的功能转换延迟。有趣的是,在缺乏MeCP2的神经元中过表达KCC2可挽救GABA功能缺陷,这表明KCC2在瑞特综合征中具有重要作用。我们进一步确定,神经元基因抑制因子RE1沉默转录因子(REST)介导MeCP2对KCC2的调控。由于KCC2是一种起效缓慢的分子,其表达水平在发育后期才达到最高,KCC2的功能缺陷可能为瑞特综合征症状延迟出现提供了解释。我们的研究表明,恢复瑞特神经元中KCC2的功能可能为瑞特综合征带来潜在的治疗方法。