Tao Deng,1,5 Vukasin M. Jovanovic,1,5 Carlos A. Tristan,1 Claire Weber,1 Pei-Hsuan Chu,1 Jason Inman,1 Seungmi Ryu,1 Yogita Jethmalani,1 Juliana Ferreira de Sousa,1 Pinar Ormanoglu,1 Prisca Twumasi,1 Chaitali Sen,1 Jaehoon Shim,2,3 Selwyn Jayakar,2,3 Han-Xiong Bear Zhang,3 Sooyeon Jo,3 Weifeng Yu,4 Ty C. Voss,1 Anton Simeonov,1 Bruce P. Bean,3 Clifford J. Woolf,2,3 and Ilyas Singeç,1,*
1National Center for Advancing Translational Sciences (NCATS), Division of Preclinical Innovation, Stem Cell Translation Laboratory (SCTL), National
Institutes of Health (NIH), Rockville, MD 20850, USA
2F.M. Kirby Neurobiology Center, Boston Children’s Hospital, Boston, MA 02115, USA
3Department of Neurobiology, Harvard Medical School, Boston, MA 02115, USA
4Sophion Bioscience, North Brunswick, NJ
Abstract
Development of new non-addictive analgesics requires advanced strategies to differentiate human pluripotent stem cells (hPSCs) into relevant cell types. Following principles of developmental biology and translational applicability, here we developed an efficient stepwise differentiation method for peptidergic and non-peptidergic nociceptors. By modulating specific cell signaling pathways, hPSCs were first converted into SOX10+ neural crest, followed by differentiation into sensory neurons. Detailed characterization, including ultrastructural analysis, confirmed that the hPSC-derived nociceptors displayed cellular and molecular 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 relevant for pain research and axonal growth/regeneration studies (e.g., TRPV1, NAV1.7, NAV1.8, TAC1, CALCA, GAP43, DPYSL2, NMNAT2). 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.
