Loss-of-function mutations in the chromodomain helicase DNA-binding 8 (CHD8) gene are strongly associated with Autism Spectrum Disorders (ASD). Indeed, the reduction of CHD8 causes transcriptional, epigenetic and cellular phenotypic changes correlated to disease, that can be monitored in assessing new therapeutic approaches. SINEUPs are a functional class of natural and synthetic antisense long non-coding RNAs able to stimulate the translation of sense target mRNA, with no effect on transcription. Here we employed synthetic SINEUP-CHD8 targeting the first and third AUG of the CHD8 coding sequence to efficiently stimulate endogenous CHD8 protein production. SINEUP-CHD8 were effective in cells with reduced levels of the target protein and in patients'-derived fibroblasts with CHD8 mutations. Functionally, SINEUP-CHD8 were able to revert molecular phenotypes associated with CHD8-suppression, i.e. genome-wide transcriptional dysregulation, and the reduction of H3K36me3 levels. Strikingly, in chd8-morpholino-treated and ENU mutant zebrafish embryos, SINEUP-chd8 injection confirmed the ability of SINEUP RNA to rescue the chd8-suppression-induced macrocephaly phenotype and neuronal hyperproliferation. Thus, SINEUP-CHD8 molecule(s) represent a proof-of-concept towards the development of a RNA-based therapy for neurodevelopmental syndromes with implications for, and beyond ASD, and relevant to genetic disorders caused by protein haploinsufficiency.

中文翻译：

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SINEUP RNA 在自闭症谱系障碍模型系统中挽救与 CHD8 抑制相关的分子表型。


染色体结构域解旋酶 DNA 结合 8 （CHD8） 基因的功能丧失突变与自闭症谱系障碍 （ASD） 密切相关。事实上，CHD8 的减少会导致与疾病相关的转录、表观遗传和细胞表型变化，这可以在评估新的治疗方法时进行监测。SINEUPs 是一类功能性的天然和合成反义长链非编码 RNA，能够刺激正义靶标 mRNA 的翻译，对转录没有影响。在这里，我们采用靶向 CHD8 编码序列的第一个和第三个 AUG 的合成 SINEUP-CHD8 来有效刺激内源性 CHD8 蛋白的产生。SINEUP-CHD8 对靶蛋白水平降低的细胞和具有 CHD8 突变的患者来源的成纤维细胞有效。在功能上，SINEUP-CHD8 能够逆转与 CHD8 抑制相关的分子表型，即全基因组转录失调和 H3K36me3 水平降低。引人注目的是，在 chd8-吗啉代处理和 ENU 突变斑马鱼胚胎中，SINEUP-chd8 注射证实了 SINEUP RNA 能够挽救 chd8 抑制诱导的大头畸形表型和神经元过度增殖。因此，SINEUP-CHD8 分子代表了开发基于 RNA 的神经发育综合征疗法的概念验证，该疗法对 ASD 有影响，并且与 ASD 引起的遗传疾病相关。