Tao Deng1, Carlos A. Tristan1, Claire Weber1, Pei-Hsuan Chu1, Seungmi Ryu1, Vukasin M. Jovanovic1, Pinar Ormanoglu1, Prisca Twumasi1, Jaehoon Shim2,3, Selwyn Jayakar2,3, Han-Xiong Bear Zhang3, Sooyeon Jo3, Ty C. Voss1, Anton Simeonov1, Bruce P. Bean3, Clifford J. Woolf2,3, Ilyas Singeç
1National Center for Advancing Translational Sciences (NCATS), Stem Cell Translation Laboratory (SCTL), National Institutes of Health (NIH), Rockville, MD, USA
2F.M. Kirby Neurobiology Center, Boston Children’s Hospital, Boston, MA 02115, USA
3Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA
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.