High-throughput screening identifies histone deacetylase inhibitors that modulate GTF2I expression in 7q11.23 microdupli
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RESEARCH
High‑throughput screening identifies histone deacetylase inhibitors that modulate GTF2I expression in 7q11.23 microduplication autism spectrum disorder patient‑derived cortical neurons Francesca Cavallo1†, Flavia Troglio2†, Giovanni Fagà3,4, Daniele Fancelli3,4, Reinald Shyti2, Sebastiano Trattaro1,2, Matteo Zanella2,10, Giuseppe D’Agostino2,11, James M. Hughes2,12, Maria Rosaria Cera3,4, Maurizio Pasi3,4, Michele Gabriele2,13, Maddalena Lazzarin1, Marija Mihailovich1,2, Frank Kooy9, Alessandro Rosa5,6, Ciro Mercurio3,4, Mario Varasi3,4 and Giuseppe Testa2,7,8*
Abstract Background: 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. Methods: 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. Results: 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.
*Correspondence: [email protected]; [email protected]; giuseppe. [email protected] † Francesca Cavallo and Flavia Troglio: shared first authorship. 2 High Definition Disease Modelling Lab: Stem Cell and Organoid Epigenetics, IEO, European Institute of Oncology IRCCS, Via Adamello 16, 20139 Milan, Italy Full list of author information is available at the end of the article © The Author(s) 2020. Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. T
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