Department of Molecular and Human Genetics, Baylor College of Medicine, Houston, Texas 77030, USA.
Jan and Dan Duncan Neurological Research Institute at Texas Children's Hospital, Houston, Texas 77030, USA.
Nature. 2015 Dec 3;528(7580):123-6. doi: 10.1038/nature16159. Epub 2015 Nov 25.
Copy number variations have been frequently associated with developmental delay, intellectual disability and autism spectrum disorders. MECP2 duplication syndrome is one of the most common genomic rearrangements in males and is characterized by autism, intellectual disability, motor dysfunction, anxiety, epilepsy, recurrent respiratory tract infections and early death. The broad range of deficits caused by methyl-CpG-binding protein 2 (MeCP2) overexpression poses a daunting challenge to traditional biochemical-pathway-based therapeutic approaches. Accordingly, we sought strategies that directly target MeCP2 and are amenable to translation into clinical therapy. The first question that we addressed was whether the neurological dysfunction is reversible after symptoms set in. Reversal of phenotypes in adult symptomatic mice has been demonstrated in some models of monogenic loss-of-function neurological disorders, including loss of MeCP2 in Rett syndrome, indicating that, at least in some cases, the neuroanatomy may remain sufficiently intact so that correction of the molecular dysfunction underlying these disorders can restore healthy physiology. Given the absence of neurodegeneration in MECP2 duplication syndrome, we propose that restoration of normal MeCP2 levels in MECP2 duplication adult mice would rescue their phenotype. By generating and characterizing a conditional Mecp2-overexpressing mouse model, here we show that correction of MeCP2 levels largely reverses the behavioural, molecular and electrophysiological deficits. We also reduced MeCP2 using an antisense oligonucleotide strategy, which has greater translational potential. Antisense oligonucleotides are small, modified nucleic acids that can selectively hybridize with messenger RNA transcribed from a target gene and silence it, and have been successfully used to correct deficits in different mouse models. We find that antisense oligonucleotide treatment induces a broad phenotypic rescue in adult symptomatic transgenic MECP2 duplication mice (MECP2-TG), and corrected MECP2 levels in lymphoblastoid cells from MECP2 duplication patients in a dose-dependent manner.
拷贝数变异经常与发育迟缓、智力残疾和自闭症谱系障碍相关。MECP2 重复综合征是男性中最常见的基因组重排之一,其特征为自闭症、智力残疾、运动功能障碍、焦虑、癫痫、反复呼吸道感染和早逝。甲基化CpG 结合蛋白 2(MeCP2)过表达引起的广泛缺陷对传统基于生化途径的治疗方法构成了严峻挑战。因此,我们寻求直接靶向 MeCP2 且适合转化为临床治疗的策略。我们首先解决的问题是,在症状出现后,神经功能障碍是否可以逆转。在一些单基因功能丧失性神经疾病模型中,包括雷特综合征中 MeCP2 的缺失,已经证明成年有症状的小鼠中的表型可以逆转,这表明,至少在某些情况下,神经解剖结构可能仍然足够完整,以至于纠正这些疾病的分子功能障碍可以恢复健康的生理学。鉴于 MECP2 重复综合征中没有神经退行性变,我们提出,在 MECP2 重复成年小鼠中恢复正常的 MeCP2 水平将挽救其表型。通过生成和表征条件性 Mecp2 过表达小鼠模型,我们在此表明,纠正 MeCP2 水平在很大程度上可以逆转行为、分子和电生理缺陷。我们还使用反义寡核苷酸策略降低 MeCP2 水平,这种策略具有更大的转化潜力。反义寡核苷酸是可以与靶基因转录的信使 RNA 选择性杂交并使其沉默的小修饰核酸,已成功用于纠正不同小鼠模型中的缺陷。我们发现,反义寡核苷酸治疗可诱导成年有症状的转基因 MECP2 重复小鼠(MECP2-TG)的广泛表型挽救,并且以剂量依赖性方式纠正 MECP2 重复患者的淋巴母细胞中的水平。