HESC-derived sensory neurons reveal an unexpected role for PIEZO2 in nociceptor mechanotransduction

Abstract

Somatosensation, the detection and transduction of external and internal stimuli, has fascinated scientists for centuries. But still, some of the mechanisms how distinct stimuli are detected and transduced are not entirely understood. Over the past decade major progress has increased our understanding in areas such as mechanotransduction or sensory neuron classification. Additionally, the accessibility to human pluripotent stem cells and the possibility to generate and study human sensory neurons has enriched the somatosensory research field.Based on our previous work, the generation of functional human mechanoreceptors, we describe here the generation of hESC-derived nociceptor-like cells. We show that by varying the differentiation strategy, we can produce different nociceptive subpopulations. One protocol in particular allowed the generation of a sensory neuron population, homogeneously expressing TRPV1, a prototypical marker for nociceptors. Accordingly, we find the cells to homogenously respond to capsaicin, to become sensitized upon inflammatory stimuli, and to respond to temperature stimulation. Surprisingly, all of the generated subtypes show mechano-nociceptive characteristics and, quite unexpectedly, loss of mechanotransduction in the absence of PIEZO2.

The Form and Function of PIEZO2

“…Long-term coexpression of NGN2 and BRN3A converts iPSCs into a subtype of cold-sensing mechanoreceptor that also relies on PIEZO2 for mechanosensitivity (90). Overexpression of NGN1 instead generates various populations of heat-sensing nociceptors (159).…”

Mechanical sensitization, increased axonal excitability, and spontaneous activity in C-nociceptors after ultraviolet B irradiation in pig skin

“…This nociceptor class was also showing a particular high expression of the voltage-sensitive sodium channel NaV1.8 and the high affinity receptor for nerve growth factor trkA. 43 Such an expression pattern would be in-line with the TTX-resistant silent nociceptors in the pig. 18 Thus, axonal excitability as assessed by slowing of conduction is highly useful for classification of C-nociceptor classes.…”

Genetic pain loss disorders

“…Fig. 295,315 . B| Differentiation of iPSCs to nociceptors using lentiviral mediated ectopic expression of neurogenin 1 (NGN1) in neural crest like cells (NCLCs).…”

“…B| Differentiation of iPSCs to nociceptors using lentiviral mediated ectopic expression of neurogenin 1 (NGN1) in neural crest like cells (NCLCs). Cells are transduced with a lentiviral vector carrying the coding sequences of NGN1, GFP and a puromycin resistence (PuroR) under the control of a doxycycline inducible promoter (TetON) 316 . C| Immunostaining of human iPSC-derived sensory neurons with β-III-tubulin (green), BRN3A (red) and Hoechst 33342 (blue).…”