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BOO induces fibrosis and EMT in urothelial cells which can be recapitulated in vitro through elevated storage and voiding pressure cycles

  • Urology - Original Paper
  • Published:
International Urology and Nephrology Aims and scope Submit manuscript

Abstract

Purpose

To determine the unique contributions from elevated voiding and storage pressures in the development of fibrosis and the epithelial-to-mesenchymal transition (EMT) in urothelial cells, and how progressive BOO pressure cycling is an important mechanical cue leading to these pathological changes.

Materials and methods

Urothelial cells isolated from control, SHAM, 2 (acute)- or 6 (chronic)-week BOO rats treated with an inflammasome inhibitor or no drug. Total RNA was isolated and RT-PCR was conducted with custom primers for pro-fibrotic and EMT genes. In separate experiments, a rat urothelial cell line was exposed to cyclic pressure regimes characteristic of acute and chronic BOO in the presence or absence of an inflammasome inhibitor. Following exposure, RT-PCR was conducted, collagen content was determined and intracellular caspase-1 activity was measured.

Results

Urothelial cells isolated from acute and chronic BOO rat models demonstrated expression of pro-fibrotic and EMT genes. Similarly, MYP3 rat urothelial cells subjected to pressure cycling regimes that reflect intravesical pressures in the acute or chronic BOO bladder also demonstrated increased expression of pro-fibrotic and EMT genes, along with elevated soluble collagen. Treatment with inflammasome inhibitors reduced expression of pro-fibrotic genes in the rat model and pressure cycling model but had a limited effect on EMT.

Conclusion

These results indicate that acute and chronic BOO pressure cycling are essential in the initiation and progression of fibrosis in the bladder via the NLRP3 inflammasome, but also provide new evidence that there is also an alternative NLRP3-independent pathway leading to EMT and fibrosis.

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Funding

Grants: NIH (R01DK103534, P20GM103444, P20GM121342), NSF (1264579).

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Authors and Affiliations

  1. Department of Bioengineering, 301 Rhodes Engineering Research Center, Clemson University, Clemson, SC, 29634-0905, USA

    Cody L. Dunton, J. Todd Purves, Francis M. Hughes Jr. & Jiro Nagatomi

  2. Division of Urology, Department of Surgery, Duke University Medical Center, Durham, NC, USA

    J. Todd Purves & Francis M. Hughes Jr.

  3. Department of Pediatrics, Duke University Medical Center, Durham, NC, USA

    J. Todd Purves

Authors
  1. Cody L. Dunton
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  2. J. Todd Purves
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  4. Jiro Nagatomi
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Contributions

CLD, JTP, FMH and JN conceived the project; CLD, JTP, FMH and JN designed the experiments; CLD and FMH performed the experiments; CLD analyzed the data; CLD, JTP, FMH and JN interpreted the results of experiments; CLD prepared the figures; CLD drafted the manuscript; CLD, JTP, FMH and JN edited and revised the manuscript CLD, JTP, FMH and JN approved the final version of the manuscript.

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Correspondence to Jiro Nagatomi.

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Dunton, C.L., Purves, J.T., Hughes, F.M. et al. BOO induces fibrosis and EMT in urothelial cells which can be recapitulated in vitro through elevated storage and voiding pressure cycles. Int Urol Nephrol 53, 2007–2018 (2021). https://doi.org/10.1007/s11255-021-02942-3

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  • DOI: https://doi.org/10.1007/s11255-021-02942-3

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