Comparative Enzymatic Profiling of Enteric vs. Cortical Neural Stem Cells: Establishing a Methodological Foundation for Modeling Enteric Neurodegeneration

Background The enteric nervous system (ENS) is increasingly implicated in the gut-brain axis pathophysiology of neurodegenerative diseases. Given the structural similarities between pathogenic amyloid-beta and industrial food protein amyloid fibrils (FPAFs), assessing enteric neurotoxicity is critical. However, current immortalized cell models fail to recapitulate specific neuro-glial vulnerabilities. Methods We established a robust primary model by isolating enteric neural stem cells (ENSCs) and cortical neural stem cells (NSCs) from E14.5 mouse embryos. Following comparable neuro-glial differentiation, we performed functional enzymatic profiling of glycolysis, the TCA cycle, and lipid metabolism. These profiles were mapped via multivariate analysis and compared alongside an Apoe knockout (KO) NSC lineage. Results ENSCs demonstrated a fundamentally distinct and more robust metabolic profile than cortical NSCs. ENSCs exhibited a potent mitochondrial-lipogenic axis—marked by significantly elevated citrate synthase, ATP-citrate lyase, and carnitine acetyltransferase—alongside a higher glycolytic flux (elevated lactate dehydrogenase). Principal Component Analysis confirmed ENSCs form a completely separate bioenergetic cluster. Furthermore, Apoe deficiency in cortical cells did not replicate this enteric phenotype but instead triggered a bioenergetic collapse and a stress-driven shift toward acetate utilization and peroxisomal oxidation. Conclusions The unique metabolic identity of the ENS disqualifies cortical or immortalized surrogates for accurate enteric modeling. This primary ENSC model, utilizing enzymatic activities as early biosensors, provides a crucial framework for evaluating the neurotoxic risks of dietary amyloidogenic proteins.