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.

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