Dataset - Dynamic persistence of intracellular bacterial communities of uropathogenic Escherichia coli in a human bladder-chip model of urinary tract infections

Abstract

Abstract Uropathogenic Escherichia coli (UPEC) is the most common causative agent of urinary tract infections and is a major cause for antibiotic prescriptions. Previous studies have shown that infection of terminally differentiated urinary bladder cells leads to the formation of intracellular bacterial communities (IBCs). However, the precise role of IBCs in recurrence of infection and antibiotic persistence, is not completely understood in part because the in situ dynamic responses of bacteria within these structures to antibiotic stress is difficult to assess in animal models. Here, we develop and characterize a human bladder-chip model of UPEC infection wherein superficial bladder epithelial cells and bladder microvascular endothelial cells are co-cultured under flow in urine and nutritive media respectively, and the mechanics of bladder filling and voiding cycles mimicked by application and release of linear strain. Time-lapse microscopy showed that infection of epithelial cells under shear stress in diluted urine led to the rapid recruitment and diapedesis of neutrophils across the endothelial-epithelial barrier and the formation of neutrophil swarms and neutrophil extracellular traps. Subsequently, two cycles of antibiotic administration interspersed with recovery periods revealed both non-growing and rapidly proliferating IBCs. Multiple stages of IBC formation captured on-chip with single-cell resolution revealed that that bacterial killing within IBCs was substantially delayed, outcomes such as shedding of bacteria and exfoliation are not mutually exclusive and rapidly reseeded the infection, and in rare instances bacterial growth in IBCs continued for the entire period of antibiotic administration. These new insights into the early stages of pathogenesis revisit the role of IBCs as harbours of persistent bacterial populations, with significant consequences for non-compliance with antibiotic regimens.