Deciphering and targeting the pathogenic circuit of nonlytic hepatitis E virus infection using macrophage-augmented organoids.

The pathophysiology caused by nonlytic viral infections is complex, often driven by macrophage-mediated immune responses that lead to hyperinflammation and collateral tissue damage. To conceptualize this complexity, we propose a pathogenic circuit comprising three interconnected nodes: nonlytic infection, inflammation, and immune-mediated cell death. To investigate this circuit, we combined hepatitis E virus (HEV), a prototypical nonlytic RNA virus, and macrophage-augmented organoids (MaugOs) as an innovative model. Here, we report successful recapitulation of the pathogenic circuit induced by HEV infection in MaugOs. Nonlytic HEV infection triggered robust inflammatory responses and subsequent cell death involving pyroptosis, apoptosis, and necroptosis pathways. By pharmacologically targeting individual circuit nodes as well as individual cell death pathways, we have dissected their interactions and identified potential therapeutic targets. Finally, we developed multitarget strategies by simultaneously targeting two or three nodes through rational drug combinations to effectively disrupt the pathogenic loop. Collectively, these findings elucidate the architecture of the pathogenic circuit underlying nonlytic HEV infection in MaugOs and inform the development of innovative multitarget therapies for improved disease treatment.