Rett syndrome (RTT) is an X-linked, neurodevelopmental disorder caused primarily by mutations in the methyl-CpG-binding protein 2 (MECP2) gene, which encodes a multifunctional epigenetic regulator with known links to a wide spectrum of neuropsychiatric disorders. Although postnatal functions of MeCP2 have been thoroughly investigated, its role in prenatal brain development remains poorly understood. Given the well-established importance of microRNAs (miRNAs) in neurogenesis, we employed isogenic human RTT patient-derived induced pluripotent stem cell (iPSC) and MeCP2 short hairpin RNA knockdown approaches to identify novel MeCP2-regulated miRNAs enriched during early human neuronal development. Focusing on the most dysregulated miRNAs, we found miR-199 and miR-214 to be increased during early brain development and to differentially regulate extracellular signal-regulated kinase (ERK)/mitogen-activated protein kinase and protein kinase B (PKB/AKT) signaling. In parallel, we characterized the effects on human neurogenesis and neuronal differentiation brought about by MeCP2 deficiency using both monolayer and three-dimensional (cerebral organoid) patient-derived and MeCP2-deficient neuronal culture models. Inhibiting miR-199 or miR-214 expression in iPSC-derived neural progenitors deficient in MeCP2 restored AKT and ERK activation, respectively, and ameliorated the observed alterations in neuronal differentiation. Moreover, overexpression of miR-199 or miR-214 in the wild-type mouse embryonic brains was sufficient to disturb neurogenesis and neuronal migration in a similar manner to Mecp2 knockdown. Taken together, our data support a novel miRNA-mediated pathway downstream of MeCP2 that influences neurogenesis via interactions with central molecular hubs linked to autism spectrum disorders.
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We thank Ting Fu, Tatiana Karadimitriou, Chuong Le, Anita Liu, Jonathan Woodson and Arooshi Kumar for excellent technical support and Chloe Delepine for advice on the manuscript, as well as additional members of the Jaenisch, Haggarty and Sur laboratories, for helpful discussion. We also thank Dr Nora Perrone-Bizzozero, Dr Andrea Allan and Dr Meilian Liu, as well as Robert Oliver, Elizabeth Solomon and Xing Zhang, for technical advice and assistance. This manuscript was supported by an NIH grant MH085802 (to MS), 2R01MH104610-15 (to RJ) and the Simons Foundation Autism Research Initiative (to MS, RJ), NIH/NIMH R01MH095088 (to SJH), Pitt-Hopkins Research Foundation (to SJH), FRAXA Research Foundation (to SJH) and Human Frontier Science Program Long-Term Fellowship (to JPKI).
The authors declare no conflict of interest.
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Mellios, N., Feldman, D., Sheridan, S. et al. MeCP2-regulated miRNAs control early human neurogenesis through differential effects on ERK and AKT signaling. Mol Psychiatry 23, 1051–1065 (2018). https://doi.org/10.1038/mp.2017.86
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