Filovirus Infection Disrupts Epithelial Barrier Function and Ion Transport in Human iPSC-Derived Gut Organoids

Gastrointestinal (GI) dysfunction, characterized by severe diarrhea and dehydration, is a central contributor to morbidity and mortality in filovirus disease in patients, yet the role of the epithelium in this clinical outcome remains poorly defined. Here, we employ induced pluripotent stem cell (iPSC)-derived human intestinal (HIOs) and colonic organoids (HCOs) to model Ebola virus (EBOV) and Marburg virus (MARV) infection. These organoids are permissive to filovirus infection and support viral replication. Bulk RNA sequencing revealed distinct intestinal and colonic epithelial responses, including apical and junctional disruption and a delayed virus-specific induction of interferon-stimulated genes. Moreover, infection impaired adenylate cyclase signaling and CFTR-mediated ion transport, providing mechanistic insight into virus- induced secretory diarrhea. This platform recapitulates key features of human GI pathology in filoviral disease and serves as a powerful system to dissect host-pathogen interactions and identify therapeutic targets. Overall design: To assess the effects of filovirus infection on the human gastrointestinal tract, we exposed induced pluripotent stem cell (iPSC)-derived human colonic organoids (HCOs) to Ebola virus (EBOV) and Marburg (MARV) virus. Bulk RNA sequencing (bulk RNA-seq) was performed on proximal HIOs (n = 3), distal HIOs (n = 3), and mock-infected controls (n = 3) at 1 and 3 days post-infection to identify transcriptomic changes. These findings were further supported by immunohistochemistry and immunofluorescence assays to validate viral infection.