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

Nat Commun. 2021 May 18;12(1):2897. 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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Differentiation / drug effects
  • Cell Differentiation / genetics
  • Cells, Cultured
  • Chromosome Deletion*
  • Chromosomes, Human, Pair 16 / genetics*
  • DNA Copy Number Variations
  • Dopaminergic Neurons / cytology
  • Dopaminergic Neurons / metabolism*
  • Dopaminergic Neurons / physiology
  • Gene Expression / drug effects
  • Humans
  • Induced Pluripotent Stem Cells / cytology
  • Induced Pluripotent Stem Cells / metabolism*
  • Nerve Net / drug effects
  • Nerve Net / metabolism*
  • Organic Chemicals / pharmacology
  • Reverse Transcriptase Polymerase Chain Reaction
  • Synaptic Transmission / drug effects
  • Synaptic Transmission / genetics*
  • rhoA GTP-Binding Protein / antagonists & inhibitors
  • rhoA GTP-Binding Protein / genetics*
  • rhoA GTP-Binding Protein / metabolism


  • Organic Chemicals
  • rhosin
  • RHOA protein, human
  • rhoA GTP-Binding Protein