Direct Induction and Functional Maturation of Forebrain GABAergic Neurons from Human Pluripotent Stem Cells

Alfred Xuyang Sun; Qiang Yuan; Shawn Tan; Yixin Xiao; Danlei Wang; Audrey Tze Ting Khoo; Levena Sani; Hoang-Dai Tran; Paul Kim; Yong Seng Chiew; Kea Joo Lee; Yi-Chun Yen; Huck Hui Ng; Bing Lim; Hyunsoo Shawn Je

doi:10.1016/j.celrep.2016.07.035

Direct Induction and Functional Maturation of Forebrain GABAergic Neurons from Human Pluripotent Stem Cells

Cell Rep. 2016 Aug 16;16(7):1942-53. doi: 10.1016/j.celrep.2016.07.035. Epub 2016 Aug 4.

Authors

Affiliations

¹ Cancer Stem Cell Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore; Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore; Department of Neurology, National Neuroscience Institute, 20 College Road, Singapore 169856, Singapore. Electronic address: [email protected].
² Molecular Neurophysiology Laboratory, Signature Program in Neuroscience and Behavioral Disorders, Duke-NUS Medical School, 8 College Road, Singapore 169857, Singapore; Graduate School for integrative Sciences and Engineering, National University of Singapore, Singapore 117456, Singapore.
³ Molecular Neurophysiology Laboratory, Signature Program in Neuroscience and Behavioral Disorders, Duke-NUS Medical School, 8 College Road, Singapore 169857, Singapore.
⁴ Cancer Stem Cell Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore.
⁵ Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore.
⁶ Department of Structure and Function of Neural Network, Korea Brain Research Institute, Daegu 701-300, Republic of Korea.
⁷ Stem Cell and Regenerative Biology, Genome Institute of Singapore, 60 Biopolis Street, Singapore 138672, Singapore; Graduate School for integrative Sciences and Engineering, National University of Singapore, Singapore 117456, Singapore.
⁸ Molecular Neurophysiology Laboratory, Signature Program in Neuroscience and Behavioral Disorders, Duke-NUS Medical School, 8 College Road, Singapore 169857, Singapore; Department of Physiology, Yong Loo Lin School of Medicine, National University of Singapore, Singapore 117597, Singapore. Electronic address: [email protected].

PMID: 27498872
DOI: 10.1016/j.celrep.2016.07.035

Abstract

Gamma-aminobutyric acid (GABA)-releasing interneurons play an important modulatory role in the cortex and have been implicated in multiple neurological disorders. Patient-derived interneurons could provide a foundation for studying the pathogenesis of these diseases as well as for identifying potential therapeutic targets. Here, we identified a set of genetic factors that could robustly induce human pluripotent stem cells (hPSCs) into GABAergic neurons (iGNs) with high efficiency. We demonstrated that the human iGNs express neurochemical markers and exhibit mature electrophysiological properties within 6-8 weeks. Furthermore, in vitro, iGNs could form functional synapses with other iGNs or with human-induced glutamatergic neurons (iENs). Upon transplantation into immunodeficient mice, human iGNs underwent synaptic maturation and integration into host neural circuits. Taken together, our rapid and highly efficient single-step protocol to generate iGNs may be useful to both mechanistic and translational studies of human interneurons.

Keywords: GABA; direct conversion; human pluripotent stem cells; interneuron; synapse.

MeSH terms

Animals
Basic Helix-Loop-Helix Transcription Factors / genetics*
Basic Helix-Loop-Helix Transcription Factors / metabolism
Biomarkers / metabolism
Cell Differentiation
Cell Line
Cerebral Cortex / cytology
Cerebral Cortex / metabolism*
Coculture Techniques
GABAergic Neurons / cytology
GABAergic Neurons / metabolism*
Gene Expression
Homeodomain Proteins / genetics
Homeodomain Proteins / metabolism
Humans
Interneurons / cytology
Interneurons / metabolism
LIM-Homeodomain Proteins / genetics
LIM-Homeodomain Proteins / metabolism
Mice
Mice, Inbred NOD
Mice, SCID
Nerve Tissue Proteins / genetics
Nerve Tissue Proteins / metabolism
Neural Stem Cells / cytology
Neural Stem Cells / metabolism
Neuroglia / cytology
Neuroglia / metabolism
Nuclear Proteins / genetics
Nuclear Proteins / metabolism
Patch-Clamp Techniques
Pluripotent Stem Cells / cytology
Pluripotent Stem Cells / metabolism*
Primary Cell Culture
Prosencephalon / cytology
Prosencephalon / metabolism*
Synapses / physiology
Synaptic Transmission / physiology
Thyroid Nuclear Factor 1
Transcription Factors / genetics
Transcription Factors / metabolism
gamma-Aminobutyric Acid / metabolism*

Substances

ASCL1 protein, human
Basic Helix-Loop-Helix Transcription Factors
Biomarkers
DLX2 protein, human
Homeodomain Proteins
LHX6 protein, human
LIM-Homeodomain Proteins
Nerve Tissue Proteins
Nuclear Proteins
Thyroid Nuclear Factor 1
Transcription Factors
gamma-Aminobutyric Acid