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

Maria Sundberg; Hannah Pinson; Richard S. Smith; Kellen D. Winden; Pooja Venugopal; Derek J.C. Tai; James F. Gusella; Michael E. Talkowski; Christopher A. Walsh; Max Tegmark; Mustafa Sahin

doi:10.1038/s41467-021-23113-z

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

Maria Sundberg, Hannah Pinson, Richard S. Smith, Kellen D. Winden, Pooja Venugopal, Derek J.C. Tai, James F. Gusella, Michael E. Talkowski, Christopher A. Walsh, Max Tegmark, Mustafa Sahin^*

^*Corresponding author for this work

Pharmacology

Research output: Contribution to journal › Article › peer-review

37 Scopus citations

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.

Original language	English (US)
Article number	2897
Journal	Nature communications
Volume	12
Issue number	1
DOIs	https://doi.org/10.1038/s41467-021-23113-z
State	Published - Dec 1 2021

Funding

We would like to thank the Flow Cytometry Core at the Hematology Oncology Division at Boston Children’s Hospital for assistance with the neural cell sorting and Dr. Lee Barrett for assistance with the image analyses at the Human Neuron Core at Boston Children’s Hospital (BCH IDDRC, U54HD090255). We are grateful to Drs. Ville Kujala, Meera Modi, Denise McGinnis, and members of the Sahin lab for critical reviewing of the manuscript and helpful comments. This study was supported by Tommy Fuss Center for Neuropsychiatric Disease Research and the Rothberg Family Fund for Cognitive Science at Boston Children’s Hospital and MIT. H.P. was supported by a fellowship from the Research Foundation Flanders (FWO-Vlaanderen) under Grant No. 11A6819N. R.S.S. was supported by F32NS100338 and K99NS112604. K.D.W. was supported by the American Academy of Neurology Neuroscience Research Training Scholarship and NIH NINDS 5K08NS112598. J.F.G., M.E.T., and D.J.C.T. received support from NIH Grants NS093200, HD096326, and GM061354. C.A.W. is an Investigator of the Howard Hughes Medical Institute. C.A.W. is supported by grants from the NINDS (R01NS032457) and NIMH (U01MH106883). M.S. is supported by NINDS R01NS113591.

ASJC Scopus subject areas

General Chemistry
General Biochemistry, Genetics and Molecular Biology
General Physics and Astronomy

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Sundberg, M., Pinson, H., Smith, R. S., Winden, K. D., Venugopal, P., Tai, D. J. C., Gusella, J. F., Talkowski, M. E., Walsh, C. A., Tegmark, M., & Sahin, M. (2021). 16p11.2 deletion is associated with hyperactivation of human iPSC-derived dopaminergic neuron networks and is rescued by RHOA inhibition in vitro. Nature communications, 12(1), Article 2897. https://doi.org/10.1038/s41467-021-23113-z

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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.",

author = "Maria Sundberg and Hannah Pinson and Smith, {Richard S.} and Winden, {Kellen D.} and Pooja Venugopal and Tai, {Derek J.C.} and Gusella, {James F.} and Talkowski, {Michael E.} and Walsh, {Christopher A.} and Max Tegmark and Mustafa Sahin",

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Sundberg, M, Pinson, H, Smith, RS, Winden, KD, Venugopal, P, Tai, DJC, Gusella, JF, Talkowski, ME, Walsh, CA, Tegmark, M & Sahin, M 2021, '16p11.2 deletion is associated with hyperactivation of human iPSC-derived dopaminergic neuron networks and is rescued by RHOA inhibition in vitro', Nature communications, vol. 12, no. 1, 2897. https://doi.org/10.1038/s41467-021-23113-z

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T1 - 16p11.2 deletion is associated with hyperactivation of human iPSC-derived dopaminergic neuron networks and is rescued by RHOA inhibition in vitro

AU - Sundberg, Maria

AU - Pinson, Hannah

AU - Smith, Richard S.

AU - Winden, Kellen D.

AU - Venugopal, Pooja

AU - Tai, Derek J.C.

AU - Gusella, James F.

AU - Talkowski, Michael E.

AU - Walsh, Christopher A.

AU - Tegmark, Max

AU - Sahin, Mustafa

PY - 2021/12/1

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N2 - 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.

AB - 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.

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

Abstract

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