iPSC-induced multilineage liver organoids, small intestinal organoids and brain organoids sustain pangenotype hepatitis E virus propagation

Background Hepatitis E virus (HEV), the leading global cause of acute viral hepatitis, lacks robust in vitro models for virology and pathogenesis research. Objective We evaluated induced pluripotent stem cell (iPSC)-induced human liver, intestinal and brain organoids (hLOs, hIOs and hBOs) as platforms for HEV infection and replication. Methods Multilineage organoids were infected with clinical HEV genotypes 1, 3 and 4. Viral tropism, host responses and antiviral efficacy were assessed. Results All organoids supported the complete life cycle of HEV. hLOs exhibited infection in hepatocytes, cholangiocytes, macrophages and stellate cells, accompanied by elevated interleukin-6 levels, impaired hepatic function (reduced secretion of albumin and Factor IX) and increased levels of alanine aminotransferase and aspartate aminotransferase, indicating hepatocellular injury. hIOs demonstrated broad epithelial (enterocyte, goblet, Paneth and endocrine) and mesenchymal infection, with disrupted barrier function (loss of tight junction proteins), proinflammatory cytokines upregulation and initiation of epithelial–mesenchymal transition. hBOs showed neuronal tropism, infecting glutamatergic, dopaminergic and GABAergic neurons, as well as astrocytes and oligodendrocytes. An increased number of TH + dopaminergic neurons was observed. Ribavirin treatment in all three models partially reversed this phenotype. Conclusion This study established iPSC-induced multilineage organoid infection models, confirming HEV’s capacity for pan-tissue infection and revealing potential pathogenic mechanisms in the liver, intestine and nervous system. This platform provides valuable tools for HEV virology research and antiviral drug development, underscoring the unique value of organoid technology in infectious disease research.