Beyond polio: advanced human in vitro models for studying non-polio enterovirus pathogenesis.

While efforts to eradicate poliovirus are ongoing, non-polio enteroviruses (NPEV) continue to pose a substantial public health threat. NPEVs cause a wide range of diseases, from mild self-limiting infections to severe respiratory, neurological, and cardiac complications, with the highest burden occurring in children and immunocompromised populations. Recent advances in human stem cell-derived cultures, organoids, air-liquid interface systems, and assembloids have transformed the study of NPEV pathogenesis by providing physiologically relevant models that recapitulate human tissue architecture, cellular complexity, and host responses. In this review, we discuss how these advanced in vitro systems have enhanced our understanding of the determinants of viral tropism, including receptor usage, epithelial polarity, tissue-specific susceptibility, and viral genetic variation. We further highlight insights into serotype-dependent differences in innate immune responses, tissue injury, and dissemination mechanisms. Finally, we explore the potential of these models for antiviral discovery and personalized therapeutic development, emphasizing their value for advancing both fundamental and translational NPEV research.