Autism spectrum disorder (ASD) is a highly prevalent neurodevelopmental condition affecting almost 1% of children, and represents a major unmet medical need with no effective drug treatment available. Duplication at 7q11.23 (7Dup), encompassing 26–28 genes, is one of the best characterized ASD-causing copy number variations and offers unique translational opportunities, because the hemideletion of the same interval causes Williams–Beuren syndrome (WBS), a condition defined by hypersociability and language strengths, thereby providing a unique reference to validate treatments for the ASD symptoms. In the above-indicated interval at 7q11.23, defined as WBS critical region, several genes, such as GTF2I, BAZ1B, CLIP2 and EIF4H, emerged as critical for their role in the pathogenesis of WBS and 7Dup both from mouse models and human studies. We performed a high-throughput screening of 1478 compounds, including central nervous system agents, epigenetic modulators and experimental substances, on patient-derived cortical glutamatergic neurons differentiated from our cohort of induced pluripotent stem cell lines (iPSCs), monitoring the transcriptional modulation of WBS interval genes, with a special focus on GTF2I, in light of its overriding pathogenic role. The hits identified were validated by measuring gene expression by qRT-PCR and the results were confirmed by western blotting. We identified and selected three histone deacetylase inhibitors (HDACi) that decreased the abnormal expression level of GTF2I in 7Dup cortical glutamatergic neurons differentiated from four genetically different iPSC lines. We confirmed this effect also at the protein level. In this study, we did not address the molecular mechanisms whereby HDAC inhibitors act on GTF2I. The lead compounds identified will now need to be advanced to further testing in additional models, including patient-derived brain organoids and mouse models recapitulating the gene imbalances of the 7q11.23 microduplication, in order to validate their efficacy in rescuing phenotypes across multiple functional layers within a translational pipeline towards clinical use. These results represent a unique opportunity for the development of a specific class of compounds for treating 7Dup and other forms of intellectual disability and autism.

中文翻译：

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高通量筛选确定组蛋白去乙酰化酶抑制剂，可调节 7q11.23 微重复自闭症谱系障碍患者源性皮层神经元中 GTF2I 的表达

自闭症谱系障碍 (ASD) 是一种非常普遍的神经发育疾病，影响了近 1% 的儿童，并且代表了一个主要的未满足的医疗需求，并且没有可用的有效药物治疗。7q11.23 (7Dup) 的重复包含 26-28 个基因，是最能描述 ASD 引起的拷贝数变异之一，并提供独特的翻译机会，因为同一区间的半缺失会导致威廉姆斯-伯伦综合征 (WBS)，由过度社交和语言强度定义的条件，从而为验证 ASD 症状的治疗提供了独特的参考。在上述定义为 WBS 关键区域的 7q11.23 区间中，一些基因，例如 GTF2I、BAZ1B、CLIP2 和 EIF4H，在小鼠模型和人体研究中对它们​​在 WBS 和 7Dup 的发病机制中的作用至关重要. 我们对 1478 种化合物进行了高通量筛选，包括中枢神经系统药物、表观遗传调节剂和实验物质，对从我们的诱导多能干细胞系 (iPSC) 队列中分化出来的患者来源的皮质谷氨酸能神经元进行了高通量筛选，监测 WBS 的转录调节间隔基因，特别关注 GTF2I，因为它具有压倒一切的致病作用。鉴定的命中通过 qRT-PCR 测量基因表达进行验证，结果通过蛋白质印迹法证实。我们鉴定并选择了三种组蛋白去乙酰化酶抑制剂 (HDACi)，它们降低了 7Dup 皮质谷氨酸能神经元中 GTF2I 的异常表达水平，这些神经元从四种基因不同的 iPSC 系分化而来。我们也在蛋白质水平上证实了这种效果。在这项研究中，我们没有解决 HDAC 抑制剂作用于 GTF2I 的分子机制。确定的先导化合物现在需要进一步在其他模型中进行进一步测试，包括患者衍生的大脑类器官和重现 7q11.23 微重复基因失衡的小鼠模型，以验证它们在拯救多个功能层的表型方面的功效在临床应用的转化管道中。这些结果代表了开发用于治疗 7Dup 和其他形式的智力障碍和自闭症的特定类别化合物的独特机会。包括患者衍生的大脑类器官和小鼠模型，重现 7q11.23 微复制的基因失衡，以验证它们在临床应用的转化管道中跨多个功能层拯救表型的功效。这些结果代表了开发用于治疗 7Dup 和其他形式的智力障碍和自闭症的特定类别化合物的独特机会。包括患者衍生的大脑类器官和小鼠模型，重现 7q11.23 微复制的基因失衡，以验证它们在临床应用的转化管道中跨多个功能层拯救表型的功效。这些结果代表了开发用于治疗 7Dup 和其他形式的智力障碍和自闭症的特定类别化合物的独特机会。