Proteomic and functional profiling of peripheral sensitization in human pluripotent stem cell-derived sensory neurons

PROJECT SUMMARY
In the dorsal root ganglia (DRG), specialized sensory neurons known as nociceptors play a crucial role in
detecting and transmitting painful stimuli. During inflammation, sensory neurons exhibit an increase in their
excitability - a form of plasticity known as peripheral sensitization that is associated with the development of
chronic pain. Though several mechanisms contributing to peripheral sensitization have been predominantly
studied in rodent DRGs, it remains unknown whether these processes also contributes to hyperexcitability in
human sensory neurons. Recently, our lab has developed a novel and scalable protocol that reproducibly
generates human pluripotent stem cell (hPSC)-derived sensory neurons. These sensory neurons exhibit key
markers found in both rodent and native DRGs, functional ion channels that regulate their excitability, and can
be activated in response to inflammatory mediators, all of which demonstrate these cells' utility in uncovering the
key signaling pathways involved in peripheral sensitization. I hypothesize that changes in de novo protein
synthesis and translation rates contribute to peripheral sensitization in hPSC-derived sensory neurons,
facilitating remodeling of the cell surface proteome and neuronal hyperexcitability. The objective of the proposed
research is to combine stem cell technology, patch-clamp electrophysiology, multi-electrode array recordings,
proximity labeling, and ribosomal and proteomic profiling to demonstrate de novo synthesis (Aim 1) and
membrane trafficking of proteins (Aim 2) are important for regulating peripheral sensitization. Furthermore, we
will also characterize whether translational rates are altered during sensitization (Aim 1) and the surface
proteome of human sensory neurons following inflammation (Aim 2). The significance of this project lies in the
potential to identify a distinct proteomic signature associated with peripheral sensitization in human DRGs, which
is highly relevant to understanding pain etiology.

Grant Number: 1FI2GM154677-01
NIH Institute/Center: NIH
Principal Investigator: David Castellano