Scalable Generation of Pseudo-Unipolar Sensory Neurons from Human Pluripotent Stem Cells

Deng, T.; Tristan, C. A.; Weber, C.; Chu, P.-H.; Ryu, S.; Jovanovic, V. M.; Ormanoglu, P.; Twumasi, P.; Shim, J.; Jayakar, S.; Zhang, H.-X. B.; Jo, S.; Voss, T. C.; Simeonov, A.; Bean, B. P.; Woolf, C. J.; Singec, I.

doi:10.1101/2022.03.24.485622

bioRxiv · 10.1101/2022.03.24.485622

Scalable Generation of Pseudo-Unipolar Sensory Neurons from Human Pluripotent Stem Cells

Deng, T.·Tristan, C. A.·Weber, C.·Chu, P.-H.·Ryu, S.·Jovanovic, V. M.·Ormanoglu, P.·Twumasi, P.·Shim, J.·Jayakar, S.·Zhang, H.-X. B.·Jo, S.·Voss, T. C.·Simeonov, A.·Bean, B. P.·Woolf, C. J.·Singec, I.

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Abstract

Development of new non-addictive analgesics requires advanced strategies to differentiate human pluripotent stem cells (hPSCs) into relevant cell types amenable for translational research. Here, we developed a highly efficient and reproducible method that differentiates hPSCs into peptidergic and non-peptidergic nociceptors. By modulating specific cell signaling pathways, hPSCs were first converted into SOX10+ neural crest cells, followed by differentiation into sensory neurons with an in vivo-like pseudo-unipolar morphology. Detailed characterization confirmed that the hPSC-derived nociceptors displayed molecular and cellular features comparable to native dorsal root ganglion (DRG) neurons, and expressed high-threshold primary sensory neuron markers, transcription factors, neuropeptides, and over 150 ion channels and receptors, including critical pain-relevant drug targets (e.g., TRPV1, TAC1, CALCA, NAV1.7, NAV1.8). Moreover, after confirming robust functional activities and differential response to noxious stimuli and specific drugs, a robotic cell culture system was employed to produce large quantities of human sensory neurons, which can be used to develop nociceptor-selective analgesics.

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Deng, T., Tristan, C. A., Weber, C., Chu, P.-H., Ryu, S., Jovanovic, V. M., Ormanoglu, P., Twumasi, P., Shim, J., Jayakar, S., Zhang, H.-X. B., Jo, S., Voss, T. C., Simeonov, A., Bean, B. P., Woolf, C. J., Singec, I.. 2022-03-27. Scalable Generation of Pseudo-Unipolar Sensory Neurons from Human Pluripotent Stem Cells. https://doi.org/10.1101/2022.03.24.485622

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