16p11.2 deletion is associated with hyperactivation of human iPSC-derived dopaminergic neuron networks and is rescued by RHOA inhibition in vitro

• Published in: Nature communications 2021-05, Vol.12 (1), p.2897-20, Article 2897
• Main Authors: Sundberg, Maria, Pinson, Hannah, Smith, Richard S., Winden, Kellen D., Venugopal, Pooja, Tai, Derek J. C., Gusella, James F., Talkowski, Michael E., Walsh, Christopher A., Tegmark, Max, Sahin, Mustafa
• Format: Article
• Language: English
• Subjects: 13/100; 14/19; 14/63; 38/91; 631/378; 631/378/1689; 631/378/2183; Biomarkers; Cell culture; Cell Differentiation - drug effects; Cell Differentiation - genetics; Cells, Cultured; Chromosome Deletion; Chromosomes, Human, Pair 16 - genetics; Copy number; CRISPR; Deletion; Disorders; DNA Copy Number Variations; Dopamine receptors; Dopaminergic Neurons - cytology; Dopaminergic Neurons - metabolism; Dopaminergic Neurons - physiology; Gene Expression - drug effects; Humanities and Social Sciences; Humans; Induced Pluripotent Stem Cells - cytology; Induced Pluripotent Stem Cells - metabolism; Inhibitory postsynaptic potentials; Mental disorders; multidisciplinary; Nerve Net - drug effects; Nerve Net - metabolism; Neural networks; Neurodevelopmental disorders; Neurons; Organic Chemicals - pharmacology; Pluripotency; Reverse Transcriptase Polymerase Chain Reaction; rhoA GTP-Binding Protein - antagonists & inhibitors; rhoA GTP-Binding Protein - genetics; rhoA GTP-Binding Protein - metabolism; RhoA protein; Science; Science (multidisciplinary); Stem cells; Synaptic Transmission - drug effects; Synaptic Transmission - genetics
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-021-23113-z

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. 16p11.2 CNVs are associated with neurodevelopmental disorders. Here, the authors show that 16p11.2 deletion is associated with hyperactivation of human iPSC-derived dopaminergic neuron networks and is rescued by RHOA inhibition in vitro.