Abstract
Reciprocal copy number variations (CNVs) of 16p11.2 are associated with a wide spectrum of neuropsychiatric and neurodevelopmental disorders. Here, we use human induced pluripotent stem cells (iPSCs)-derived dopaminergic (DA) neurons carrying CNVs of 16p11.2 duplication (16pdup) and 16p11.2 deletion (16pdel), engineered using CRISPR-Cas9. We show that 16pdel iPSC-derived DA neurons have increased soma size and synaptic marker expression compared to isogenic control lines, while 16pdup iPSC-derived DA neurons show deficits in neuronal differentiation and reduced synaptic marker expression. The 16pdel iPSC-derived DA neurons have impaired neurophysiological properties. The 16pdel iPSC-derived DA neuronal networks are hyperactive and have increased bursting in culture compared to controls. We also show that the expression of RHOA is increased in the 16pdel iPSC-derived DA neurons and that treatment with a specific RHOA-inhibitor, Rhosin, rescues the network activity of the 16pdel iPSC-derived DA neurons. Our data suggest that 16p11.2 deletion-associated iPSC-derived DA neuron hyperactivation can be rescued by RHOA inhibition.
Highlights
Reciprocal copy number variations (CNVs) of 16p11.2 are associated with a wide spectrum of neuropsychiatric and neurodevelopmental disorders
We investigated the functional and genetic characteristics of the DA neurons derived from isogenic human induced pluripotent stem cells (iPSCs) lines with CRISPR-Cas9induced 16p11.2 CNVs30
Our findings indicate that the KCTD13 pathway and Ras homolog family member A (RHOA) activation have significant roles in the hyperactivation of DA neuron networks in neuropsychiatric disorders with 16pdel, and the inhibition of this signaling pathway can serve as a therapeutic target for these disorders
Summary
Reciprocal copy number variations (CNVs) of 16p11.2 are associated with a wide spectrum of neuropsychiatric and neurodevelopmental disorders. Patients with 16pdel have an increased risk of developmental delay and ASD5, and behavioral problems associated with this CNV include attention deficit hyperactivity disorder[6] Another meta-analysis of 6882 SCZ patients and 6316 controls determined that 0.35% of the patients had 16pdup, as compared with a 0.03% in the control population[2]. Induced pluripotent stem cell (iPSC)-derived cortical neurons with CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-Cas[9] induced loss of KCTD13 display deficits in neurite formation and network activity[14] While these studies indicate that the KCTD13 pathway may have a central role in the formation of functional cortical neuron networks in 16p11.2 disorders like autism and SCZ, its effects on development of other neuron subtypes have not been studied in detail. Our findings indicate that the KCTD13 pathway and Ras homolog family member A (RHOA) activation have significant roles in the hyperactivation of DA neuron networks in neuropsychiatric disorders with 16pdel, and the inhibition of this signaling pathway can serve as a therapeutic target for these disorders
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