TY - JOUR TI - Dynamic persistence of UPEC intracellular bacterial communities in a human bladder-chip model of urinary tract infection AU - Sharma, Kunal AU - Dhar, Neeraj AU - Thacker, Vivek V AU - Simonet, Thomas M AU - Signorino-Gelo, Francois AU - Knott, Graham W AU - McKinney, John D A2 - Helaine, Sophie A2 - Storz, Gisela A2 - Radisic, Milica VL - 10 PY - 2021 DA - 2021/07/05 SP - e66481 C1 - eLife 2021;10:e66481 DO - 10.7554/eLife.66481 UR - https://doi.org/10.7554/eLife.66481 AB - Uropathogenic Escherichia coli (UPEC) proliferate within superficial bladder umbrella cells to form intracellular bacterial communities (IBCs) during early stages of urinary tract infections. However, the dynamic responses of IBCs to host stresses and antibiotic therapy are difficult to assess in situ. We develop a human bladder-chip model wherein umbrella cells and bladder microvascular endothelial cells are co-cultured under flow in urine and nutritive media respectively, and bladder filling and voiding mimicked mechanically by application and release of linear strain. Using time-lapse microscopy, we show that rapid recruitment of neutrophils from the vascular channel to sites of infection leads to swarm and neutrophil extracellular trap formation but does not prevent IBC formation. Subsequently, we tracked bacterial growth dynamics in individual IBCs through two cycles of antibiotic administration interspersed with recovery periods which revealed that the elimination of bacteria within IBCs by the antibiotic was delayed, and in some instances, did not occur at all. During the recovery period, rapid proliferation in a significant fraction of IBCs reseeded new foci of infection through bacterial shedding and host cell exfoliation. These insights reinforce a dynamic role for IBCs as harbors of bacterial persistence, with significant consequences for non-compliance with antibiotic regimens. KW - urinary tract infections KW - organ-on-chip KW - antibiotics KW - neutrophil extracellular traps KW - intracellular bacterial communities KW - time-lapse microscopy JF - eLife SN - 2050-084X PB - eLife Sciences Publications, Ltd ER -