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Human bladder organoids model urinary tract infection and bacteriophage therapy

Zulk, J. J.; Robertson, C. M.; Ottinger, S.; Kambal, A.; Tostado, A. R.; Fleck, R. C.; Shea, A. E.; Coarfa, C.; Blutt, S. E.; Maresso, A. W.; Patras, K. A.

2025-07-30 microbiology
10.1101/2025.07.30.667685 bioRxiv
Show abstract

Urinary tract infections (UTIs), primarily caused by uropathogenic Escherichia coli (UPEC), are among the most common antibiotic-resistant infections. Despite this, currently available preclinical UTI models lack the breadth of morphotypic and heterogenous cell populations of the human bladder, impairing the development of novel therapies. To address these limitations, we developed human bladder organoids derived from the bladder stem cells of multiple healthy donors which recapitulate cellular diversity of the urothelium. Using bulk and single cell RNA-sequencing, we characterized organoid responses to UPEC and phage exposure individually and in combination to model phage therapy. Although phage minimally affected the uroepithelium in the absence of infection, during UTI, phage treatment reduced bacterial burdens and dampened inflammatory responses and barrier disruption. Collectively, our findings highlight human bladder organoids as a tool for capturing conserved and individual-specific uroepithelial responses to infection while also providing preclinical efficacy and safety testing for therapeutic development.

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