Fluid flow sensitizes bacterial pathogens to chemical stress

Padron, G. C.; Shuppara, A. M.; Sharma, A.; Koch, M. D.; Palalay, J.-J. S.; Radin, J. N.; Kehl-Fie, T. E.; Imlay, J. A.; Sanfilippo, J. E.

doi:10.1101/2022.09.07.506966

bioRxiv · 10.1101/2022.09.07.506966

Fluid flow sensitizes bacterial pathogens to chemical stress

Padron, G. C.·Shuppara, A. M.·Sharma, A.·Koch, M. D.·Palalay, J.-J. S.·Radin, J. N.·Kehl-Fie, T. E.·Imlay, J. A.·Sanfilippo, J. E.

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Abstract

Cells regularly experience fluid flow in natural systems. However, most experimental systems rely on batch cell culture and fail to consider the effect of flow-driven dynamics on cell physiology. Using microfluidics and single-cell imaging, we discover that the interplay of physical shear rate (a measure of fluid flow) and chemical stress trigger a transcriptional response in the human pathogen Pseudomonas aeruginosa. In batch cell culture, cells protect themselves by quickly scavenging the ubiquitous chemical stressor hydrogen peroxide (H2O2) from the media. In microfluidic conditions, we observe that cell scavenging generates spatial gradients of H2O2. High shear rates replenish H2O2, abolish gradients, and generate a stress response. Combining mathematical simulations and biophysical experiments, we find that cells in flow are sensitive to a H2O2 concentration that is 100-1000 times lower than traditionally studied in batch cell culture. Surprisingly, the shear rate and H2O2 concentration required to trigger a transcriptional response closely match their respective values in the human bloodstream. Thus, our results explain a long-standing discrepancy between H2O2 levels in experimental and natural systems. Finally, we demonstrate that the shear rate and H2O2 concentration found in the human bloodstream trigger gene expression in the blood-relevant human pathogen Staphylococcus aureus, suggesting that flow sensitizes bacteria to chemical stress in natural environments.

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Padron, G. C., Shuppara, A. M., Sharma, A., Koch, M. D., Palalay, J.-J. S., Radin, J. N., Kehl-Fie, T. E., Imlay, J. A., Sanfilippo, J. E.. 2022-09-07. Fluid flow sensitizes bacterial pathogens to chemical stress. https://doi.org/10.1101/2022.09.07.506966

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