Associated Director FUJIFILM Cellular Dynamics Madison, Wisconsin, United States
Abstract: With the recent FDA approval of Suzetrigine, Vertex’s selective pain inhibitor targeting NaV1.8 sodium channels, there is renewed biopharma focus on non-opioid based pain drugs. The desire to improve predictability is fueling interest in new human-relevant models, with a strong focus on induced pluripotent stem cell (iPSC)-derived peripheral sensory neurons. These cells have the potential to advance pain research, however, a deeper understanding of how they recapitulate relevant expression and function of non-opioid pain targets is needed. In this study, baseline transcriptomic characterization of a novel human iPSC-derived sensory neurons, iCell® Sensory Neurons (FUJIFILM Cellular Dynamics), was performed and compared to human dorsal root ganglion (hDRG) and alternative published iPSC-derived sensory neuron protocols. The bulk RNAseq data demonstrated that iPSC-based methods yielded cells that clustered distinctly from hDRG. Within this grouping of iPSC-derived sensory neurons, subclusters were identified, suggesting that variations in differentiation protocols can impact the resulting sensory neuron populations. Importantly, iCell Sensory Neurons showed the highest expression of non-opioid pain-relevant genes (i.e., SCN9A, SCN10A, TRPV1, PEIZO2, and P2RX3) which was corroborated with single cell RNA-seq data showing these genes were expressed in the majority of the population. In addition, these sensory neurons also displayed accentuated expression of neural maturation markers, calcium handing genes, sodium channels, and potassium channels, as compared to cells from other iPSC differentiation protocols. These genomic data correlate with functional data in cell-based assays, including robust calcium influx responses to sensory-specific molecules (i.e., capsaicin or Yoda2) and relevant electrophysiology (low spontaneous activity and sensory-induced activity) on MEA. In summary, iPSC-derived sensory neurons have a transcriptome profile relevant for human in vitro modeling of pain and neuropathy. Importantly, iCell Sensory Neurons express premier non-opioid pain targets within a population that is enriched in ion channels and intracellular signaling pathways that correlate with advanced sensory function.
