Parasympathetic neurons derived from human pluripotent stem cells model human diseases and development
Parasympathetic neurons (parasymNs) belong to the autonomic nervous system (ANS) and are critical for unconscious body responses, including rest-and-digest and calming the body. ParasymN dysfunction has been found involved in neural diseases such as autonomic neuropathy and neurological autoimmune disease; and parasymN innervation is important for organ development. However, human parasymN function and dysfunction is vastly understudied, due to the lack of a model system. Human pluripotent stem cell (hPSC)-derived neurons can fill this void and serve for disease modeling, drug screening, and transplantation therapy. Here, we developed a differentiation paradigm detailing for the first time, the derivation of functional human parasymNs from Schwann cell progenitors (SCP). We employ these neurons (i) to assess human ANS development paradigms, (ii) to model neuropathy in the genetic disorder Familial Dysautonomia (FD), (iii) to show parasymN dysfunction during SARS-CoV-2 infection, (iv) to model the autoimmune disease Sjörgen's syndrome and, (v) to show that parasymN innervation of white adipocytes during development and help mature the tissue. Our model system will become instrumental for future disease mechanistic and drug discovery studies as well as for human developmental studies. Overall design: To understand the development of parasympathetic neurons, their function, and their missregulation in diseases, we developed a protocol to differentiate parasympathetic neurons from human pluripotent stem cells. To characterize their transcriptome at different developmental stages, we collected RNA on different days throuhgout the differentiation (day 10, 14, 16, and 30) from three biological replicates (independent differentiations). We also collected RNA from parasympathetic neurons (day 30) differentiated from iPSCs dervied from one Familial dysautonomia patient (S2). We performed RNAseq of each sample. To characterize their cellular composition, we collected single cell RNA data on day 30 from two independent differentiations (biological replicates).