TY - JOUR
T1 - Regulatory networks specifying cortical interneurons from human embryonic stem cells reveal roles for CHD2 in interneuron development
AU - Meganathan, Kesavan
AU - Lewis, Emily M.A.
AU - Gontarz, Paul
AU - Liu, Shaopeng
AU - Stanley, Edouard G.
AU - Elefanty, Andrew G.
AU - Huettner, James E.
AU - Zhang, Bo
AU - Kroll, Kristen L.
N1 - Funding Information:
ACKNOWLEDGMENTS. We thank Matheus Victor and Andrew Yoo for assisting in cIN transplantations and for the SYN–GFP plasmid; David Mu for the NKX2-1 vector (Addgene); and Andrew Yoo and Harrison W. Gabel for constructive suggestions on the manuscript. This project was supported by NIH Grants GM66815 (to K.L.K.) and NS3088 (to J.E.H.); March of Dimes Grant 1-FY13-413 (to K.L.K.); grants from the American Epilepsy Society, the McDonnell Center for Cellular and Molecular Neurobiology at Washington University (WU), and the WU Center for Regenerative Medicine (CRM); WU Institute of Clinical and Translational Sciences Grant UL1 TR000448; the WU Animal Surgery Core; and the WU Gene Technology Access Center (supported by Siteman Cancer Center Grants CA91842 and UL1 TR000448). K.M. was supported by a Postdoctoral Fellowship from the WU CRM, and E.M.A.L. was supported by a National Institute of General Medical Science T32 Training Grant.
PY - 2017/12/26
Y1 - 2017/12/26
N2 - Cortical interneurons (cINs) modulate excitatory neuronal activity by providing local inhibition. During fetal development, several cIN subtypes derive from the medial ganglionic eminence (MGE), a transient ventral telencephalic structure. While altered cIN development contributes to neurodevelopmental disorders, the inaccessibility of human fetal brain tissue during development has hampered efforts to define molecular networks controlling this process. Here, we modified protocols for directed differentiation of human embryonic stem cells, obtaining efficient, accelerated production of MGE-like progenitors and MGE-derived cIN subtypes with the expected electrophysiological properties. We defined transcriptome changes accompanying this process and integrated these data with direct transcriptional targets of NKX2-1, a transcription factor controlling MGE specification. This analysis defined NKX2-1-associated genes with enriched expression during MGE specification and cIN differentiation, including known and previously unreported transcription factor targets with likely roles in MGE specification, and other target classes regulating cIN migration and function. NKX2-1-associated peaks were enriched for consensus binding motifs for NKX2-1, LHX, and SOX transcription factors, suggesting roles in coregulating MGE gene expression. Among the NKX2-1 direct target genes with cIN-enriched expression was CHD2, which encodes a chromatin remodeling protein mutated to cause human epilepsies. Accordingly, CHD2 deficiency impaired cIN specification and altered later electrophysiological function, while CHD2 coassociated with NKX2-1 at cis-regulatory elements and was required for their transactivation by NKX2-1 in MGE-like progenitors. This analysis identified several aspects of gene-regulatory networks underlying human MGE specification and suggested mechanisms by which NKX2-1 acts with chromatin remodeling activities to regulate gene expression programs underlying cIN development.
AB - Cortical interneurons (cINs) modulate excitatory neuronal activity by providing local inhibition. During fetal development, several cIN subtypes derive from the medial ganglionic eminence (MGE), a transient ventral telencephalic structure. While altered cIN development contributes to neurodevelopmental disorders, the inaccessibility of human fetal brain tissue during development has hampered efforts to define molecular networks controlling this process. Here, we modified protocols for directed differentiation of human embryonic stem cells, obtaining efficient, accelerated production of MGE-like progenitors and MGE-derived cIN subtypes with the expected electrophysiological properties. We defined transcriptome changes accompanying this process and integrated these data with direct transcriptional targets of NKX2-1, a transcription factor controlling MGE specification. This analysis defined NKX2-1-associated genes with enriched expression during MGE specification and cIN differentiation, including known and previously unreported transcription factor targets with likely roles in MGE specification, and other target classes regulating cIN migration and function. NKX2-1-associated peaks were enriched for consensus binding motifs for NKX2-1, LHX, and SOX transcription factors, suggesting roles in coregulating MGE gene expression. Among the NKX2-1 direct target genes with cIN-enriched expression was CHD2, which encodes a chromatin remodeling protein mutated to cause human epilepsies. Accordingly, CHD2 deficiency impaired cIN specification and altered later electrophysiological function, while CHD2 coassociated with NKX2-1 at cis-regulatory elements and was required for their transactivation by NKX2-1 in MGE-like progenitors. This analysis identified several aspects of gene-regulatory networks underlying human MGE specification and suggested mechanisms by which NKX2-1 acts with chromatin remodeling activities to regulate gene expression programs underlying cIN development.
KW - CHD2
KW - Cortical interneurons
KW - Human embryonic stem cells
KW - Medial ganglionic eminence
KW - NKX2-1
UR - http://www.scopus.com/inward/record.url?scp=85039729973&partnerID=8YFLogxK
U2 - 10.1073/pnas.1712365115
DO - 10.1073/pnas.1712365115
M3 - Article
C2 - 29229852
AN - SCOPUS:85039729973
SN - 0027-8424
VL - 114
SP - E11180-E11189
JO - Proceedings of the National Academy of Sciences of the United States of America
JF - Proceedings of the National Academy of Sciences of the United States of America
IS - 52
ER -