A call for a unified and multimodal definition of cellular identity in the enteric nervous system

The enteric nervous system (ENS), the largest division of autonomic nervous system, is a tantalizing frontier in neuroscience. With the advent of single-cell transcriptomics, the ENS has been increasingly well-characterized. Precise functional mapping of enteric neuron diversity is critical for understanding ENS biology and disease, but technical barriers remain. We used different approaches to compare and contrast functional annotations of several independently-reported ENS datasets. Differential module scoring, co-expression and correlation analysis, unbiased biological function hierarchical clustering, data integration and label transfer highlighted substantial discrepancies stemming from an overreliance on transcriptomics data without adequate tissue validations. For understanding enteric neurons’ functional identity, it is imperative to expand tissue sources and incorporate technologies such as multiplexed imaging, electrophysiology, spatial transcriptomics, as well as comprehensive epigenome, proteome, and metabolome profiling. Harnessing human pluripotent stem cell models provides unique opportunities for ENS lineage tracing and offers unparalleled scalability and amenability to genetic and functional screens. We encourage a paradigm shift in our comprehension of ENS cellular and functional complexity by calling for large-scale collaborations and research investments.