Modelling pain states with hiPSC derived sensory neurons

We report the development of a sensory neuron differentiation protocol which can be used to produce populations of nociceptive sensory neurons. Following transcriptomic and electrophysiological characterisation, we used cultures to investigate molecular pain pathways by exposing cells to three pain related stimuli: heat, cold and a migraine model. Samples were taken at four time points, up to 18 hours after stimulus exposure. Bulk RNA sequencing of samples showed that for temperature models, gene expression changes were dominated by those involved in cell stress and cell death mechanisms. For the migraine model, we obseved a complex pattern of exposure-associated gene expression changes, including the up-regulation of several genes and pathways involved in neuronal and synaptic function. We processed 32 samples in total. Three replicates were taken for each treatment type at each exposure time point. Heat stimulus = 45degC; cold stimulus = 4degC; migraine stimulus = 10uM bradykinin, 1uM histamine, 1uM prostaglandin E2 and 500nM serotonin. Exposure times were Short = 20 mins, Mid = 50 mins, Long = 90 mins and Overnight = 18 hours.