Recapitulating dengue virus infection with human pluripotent stem cell-derived liver organoids for antiviral screening

Dengue virus (DENV) poses a major global health threat, affecting an estimated 100 to 400 million people annually. The infection and pathogenesis remain incompletely understood, and no antiviral drug is currently approved for DENV treatment. Here, we develop a human pluripotent stem cell (hPSC)-derived liver organoid (hPLO) model to characterize DENV infection and screen for antivirals. The hPLOs, containing various liver cell types, are highly susceptible to DENV-2 infection, resulting in severe cell death and morphological changes that mimic the pathology observed in severe dengue cases. Single-cell RNA sequencing (scRNA-seq) of DENV-2 infected hPLOs reveals that proliferating hepatocyte-like cells are the primary target cells of DENV-2, with significant mitochondrial damage and alterations in cell-type composition. Further drug screening in hPLOs identifies oxyresveratrol (Oresveratrol, ORES) and omaveloxolone (RTA 408) as potent anti-DENV candidates. These compounds enhance resistance to DENV-2 infection by activating the NRF2 pathway, reducing oxidative stress, and preserving mitochondrial function. The efficacy of ORES and RTA 408 is further validated in the established AG6 mouse model. Our study not only establishes hPLOs as a valuable platform for studying DENV infection and pathogenesis, but also highlights the vital role of NRF2-mediated mitochondrial function for antiviral development.