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NIH Research Festival

September 12 – 14, 2018

Conversion of human pluripotent stem cells to sensory neurons by transcriptional programming

Thursday, September 13, 2018 – Poster Session IV
3:30 – 5:00 p.m.

FAES Terrace

NINDS

STEMCELL-4

Authors

  • AR Nickolls
  • MM Lee
  • R Lam
  • M Szczot
  • D Espinoza
  • ME Ward
  • AT Chesler
  • CG Bonnemann

Abstract

The in vitro production of peripheral sensory neurons from pluripotent stem cells has been traditionally accomplished by complex induction protocols, often ineffective and time-consuming. To overcome these obstacles, we developed a simplified system to rapidly and efficiently generate human induced sensory neurons. We found that forced expression of two transcription factors, NGN2 and BRN3A, rapidly and efficiently converts human pluripotent stem cells into peripheral sensory neurons. Within seven days, >90% of cells adopted neuronal morphology and expressed both pan-neuronal and sensory-specific markers. Further, we detected mRNA transcripts for a variety of receptor genes that bestow particular sensory functions in peripheral neurons, including TRPM8, TRPA1, P2RX3, and notably PIEZO2. Given PIEZO2’s role as a mechanotransduction channel, we used a micromanipulator probe to indent the cell membrane while measuring electrical responses. Mechanical stimulation elicited robust excitatory currents in the converted cells, a hallmark of mechanosensory neurons in vivo. We applied this technique to neurons derived from patients with PIEZO2 deficiency syndrome – a condition characterized by profound loss of proprioception and touch sensation. Remarkably, assessment of PIEZO2-deficient neurons revealed a complete absence of mechanically activated currents. Collectively, these data confirm PIEZO2’s essential role in human mechanosensation and demonstrate loss-of-function mutations with profound impact on sensory neuron physiology. This streamlined model system may broadly serve as a tool in high-throughput screens for drug discovery and identifying molecular processes involved in peripheral nervous system disorders.

Scientific Focus Area: Stem Cell Biology

This page was last updated on Friday, March 26, 2021

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