Transcriptional Programming of Human Mechanosensory Neuron Subtypes from Pluripotent Stem Cells

In vitro generation of human peripheral sensory neurons may provide a framework for novel drug screening platforms and disease models of touch and pain. However, derivation of a functionally pure sensory neuron population remains a major unmet challenge. We discovered that, by expressing the transcription factors NGN2 and BRN3A, human pluripotent stem cells can be induced to differentiate into a surprisingly homogenous culture of cold- and mechano-sensing neurons. Although such a neuronal subtype has not been reported in mice, we found molecular evidence of its existence in adult human sensory ganglia. Combining NGN2 and BRN3A programming with neural crest patterning, we produced two additional populations of sensory neurons, including a more specialized mechanosensory neuron subtype. Finally, we applied this system to model a rare inherited sensory disorder, characterized by profound impairment of touch sensation and proprioception, caused by inactivating mutations in PIEZO2. Together these findings establish an approach to specify distinct sensory neuron subtypes in vitro, underscoring the utility of stem cell technology to capture human-specific features of physiology and disease. Overall design: RNA sequencing was performed on a total of 13 human samples. Of these samples, one represented a pool of human Dorsal Root Ganglia (hDRG), three represented human undifferentiated induced pluripotent stem cells (iPSC), three represented sensory neurons induced by NGN2-BRN3A expression in iPSCs (iSN), three represented sensory neurons induced by long term NGN2-BRN3A expression in iPSC-derived neural crest (NC_iSN1), and three represented sensory neurons induced by short term NGN2-BRN3A expression in iPSC-derived neural crest (NC_iSN2).