(1151-D) Large-scale Generation of Human iPSC-derived Sensory Neurons for Pain Research

Abstract: Sensory neurons of the peripheral somatic nervous system respond to a variety of impulses from sensory organs including touch, position in space (proprioception), temperature, and pain. Sensory neurons are of great value in pain research to aid in the development of better chemotherapy drugs with manageable side effects such as chemotherapy-induced peripheral neuropathy (CIPN) and hematologic toxicity. Differentiating human induced pluripotent stem cells (iPSC) enables access to authentic human sensory neurons, but challenges arise when attempting to consistently produce large quantities of these cells. Here we report the large-scale, directed differentiation of human sensory neurons from two (2) different iPSC lines, including both male and female donors. These iPSC-derived sensory neurons have high purity (>80% BRN3A+/UCHL1+) and express hallmark nociceptive channels (i.e., Nav1.7 and Nav1.8) and receptors (i.e., TRPV1 and P2RX). We characterized sensory function of these neurons using electrophysiology and calcium imaging. Notably, we confirmed the presence of P2RX and TRP channels and their response to different stimuli including capsaicin, menthol, and ATP using calcium imaging. Additionally, these sensory neurons display cytotoxicity to chemotherapeutic drugs (i.e., paclitaxel) in a dose-dependent manner and release substance-P and CGRP peptides in response to pain mediators. These data demonstrate a process for robustly generating iPSC-derived sensory neurons across different iPSC donor lines that result in expression of characteristic markers and respond appropriately to known pharmacology in functional assays. Large-scale production of such human sensory neurons will pave the road for studying sensory neuron properties and has the potential for use in high-throughput drug screening for pain research.