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Castration immunoregulates toll-like receptor-4 in male bladder cancer

  • Urology - Original Paper
  • Published:
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Abstract

Purpose

Among diverse Pattern Recognition Receptors (PRRs), Toll-like receptor-4 (TLR-4) is a key urothelial trigger for innate immune response impacting urothelial bladder carcinoma (BC). Androgen activation promotes immunotolerance, playing an immunoregulatory role by unknown mechanisms. We explored the castration impact on urothelial TLR-4 modulation in carcinogenesis and immunotherapeutic scenario.

Methods

Intact (SHAM) versus castrated male Fisher-344 rats were evaluated in 2 scenarios: (A) Carcinogenesis: After randomization to SHAM (n = 5) and Castration (n = 5), carcinogenesis was induced by four intravesical doses of 1.5 mg/kg n-methyl-n-nitrosourea (MNU) every 15 days. (B) Treatment: After ultrasonographic confirmed MNU-induced papillary BC on week 8, rats were randomized to SHAM (n = 5) and Castration (n = 5) and offered 6 weekly intravesical treatment of 106 CFU of bacillus Calmette Guerin (BCG) in 0.2 ml saline. After 15 weeks the urinary bladders underwent histopathology. Urothelial cell proliferation was measured by Ki-67 immunohistochemistry (IHC), and TLR-4 expression was quantified by IHC and WB.

Results

Castration induced higher TLR-4 urothelial expression (p = 0.007) and anticarcinogenic effect with fewer urothelial tumors (60 vs. 80%) and lower urothelial cell proliferation compared to intact animals (p = 0.008). In the intravesical BCG treatment setting, castration has potentialized the BCG activation of TLR-4 (p = 0.007) with no residual in situ carcinoma compared to intact animals, suggesting the potential to amplify the BCG immune response.

Conclusion

To our knowledge, this is the first description of TLR-4 urothelial expression hormonal modulation. The described castration-mediated immunomodulation will help to improve the knowledge of urothelial cancer gender diversities and PRRs modulations with treatment implications.

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Acknowledgements

We would like to thank the expert uropathologist, Prof. Athanase Billis for the urothelial tumors’ classification.

Funding

Ministério da Ciência e Tecnologia/Conselho Nacional de Desenvolvimento Científico e Tecnológico—CNPq, processo 304747/2018–1.

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Authors and Affiliations

  1. UroScience, School of Medical Sciences, State University of Campinas, UNICAMP, Campinas, SP, Brazil

    Leonardo Oliveira Reis, Ana Clara Ciglioni Salustiano, Diego Moreira Capibaribe, Isis Gallani Avansini Kiehl & Fernandes Denardi

  2. Pontifical Catholic University of Campinas, PUC-Campinas, Sao Paulo, Brazil

    Leonardo Oliveira Reis

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  1. Leonardo Oliveira Reis
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  2. Ana Clara Ciglioni Salustiano
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  3. Diego Moreira Capibaribe
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  4. Isis Gallani Avansini Kiehl
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Contributions

RLO: Protocol/project development, Data analysis, Manuscript writing/editing. Salustiano ACC and Kiehl IGA: Data collection/management/analysis. Capibaribe DM, Denardi F: Data analysis.

Corresponding author

Correspondence to Leonardo Oliveira Reis.

Ethics declarations

Conflicts of interest

None.

Research involving human participants and/or animals

Local Animal Research Ethics Committee Approval no 4541–1/2017 (CEUA/UNICAMP).

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Not applicable.

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Supplementary Information

Below is the link to the electronic supplementary material.

11255_2022_3336_MOESM1_ESM.tiff

Supplementary Fig. 1: Study design: groups, interventions, and weeks of protocol (b: bladder, p: prostate) (TIFF 2377 KB)

11255_2022_3336_MOESM2_ESM.pdf

Supplementary Fig. 2: Representative bladder urothelial cancer pTa, pT1 low-grade and pT1 high-grade in the male counterpart of Fisher 344 immunocompetent bladder cancer model (PDF 3576 KB)

11255_2022_3336_MOESM3_ESM.tiff

Supplementary Fig. 3. Protein quantification of TLR-4 in the bladder, Western Blotting. Different letters denote p < 0.05 (TIFF 2377 KB)

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Reis, L.O., Salustiano, A.C.C., Capibaribe, D.M. et al. Castration immunoregulates toll-like receptor-4 in male bladder cancer. Int Urol Nephrol 54, 2845–2853 (2022). https://doi.org/10.1007/s11255-022-03336-9

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  • DOI: https://doi.org/10.1007/s11255-022-03336-9

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