Urothelial genotoxicity of household chemicals in healthy canine urinary bladder organoids relative to observed urinary exposures in pet dogs - Research Article - Genotoxicity thresholds and in vitro organoid findings

Sep 29, 2026

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Study characterizes urothelial genotoxicity of the bladder carcinogens acrolein, inorganic arsenic, and 2,6-dimethylaniline (2,6-DMA) using a novel in vitro canine urinary bladder organoid model, with alkaline CometChip assays measuring DNA strand breaks and oxidative DNA damage. Genotoxic thresholds were established for each chemical, then compared with urinary concentrations reported in prior case-control surveys of pet dogs with and without urothelial carcinoma. Results indicate genotoxic urinary concentrations for acrolein (20 uM) and 2,6-DMA (0.01 uM) in subsets of dogs, while inorganic arsenic showed none at the single measured time point.

LanguageEnglish

CategoryResearch & Report

FormatPDF

Pages8

Reading Time20 min

AccessFree to download

Introduction

Study aim and relevance to canine urothelial carcinoma

Background on urinary exposures and prior genotoxicity evidence

Methods

Canine urinary bladder organoids

Results

Genotoxicity thresholds for acrolein, inorganic arsenic, and 2,6-DMA

Conclusion

Implications for household exposures and future assessment

TYPE Original Research PUBLISHED 23 December 2025 DOI 10.3389/fvets.2025.1702980 OPEN ACCESS EDITED BY Zhigang Zhang, Northeast Agricultural University, China REVIEWED BY Saurabh Dilip Bhandare, Foxabell-Laboratorium Investigativum,“Laboratorium Scientiae et Studiorum Investigativorum”, India Maria Malvina Tsamouri, Athos Therapeutics, United States *CORRESPONDENCE Lauren A. Trepanier [lauren.trepanier@wisc.edu](lauren.trepanier@wisc.edu) RECEIVED 10 September 2025 REVISED 10 November 2025 ACCEPTED 24 November 2025 PUBLISHED 23 December 2025 CITATION Peterson HM, Zdyrski C, Allenspach K, Mochel JP and Trepanier LA (2025) Urothelial genotoxicity of household chemicals in healthy canine urinary bladder organoids relative to observed urinary exposures in pet dogs. Front. Vet. Sci. 12:1702980 . doi: 10.3389/fvets.2025.1702980 COPYRIGHT © 2025 Peterson, Zdyrski, Allenspach, Mocheland Trepanier. This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY) . The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. Urothelial genotoxicity of household chemicals in healthy canine urinary bladder organoids relative to observed urinary exposures in pet dogs Hannah M. Peterson 1, Christopher Zdyrski 2,3, Karin Allenspach 2,3, Jonathan P. Mochel 2,3,4 and Lauren A. Trepanier 1* 1Department of Medical Sciences, University of Wisconsin-Madison, Madison, WI, United States, 2Department of Pathology, University of Georgia, Athens, Georgia, 3Precision One Health Initiative, University of Georgia, Athens, Georgia, 4Office of Biotechnology, Iowa State University, Ames, IA, United States Introduction: Urothelial carcinoma (UC) in pet dogs closely resembles human muscle-invasive UC, which is associated with environmental chemical carcinogens. The aim of this study was to determine whether urinary concentrations of the bladder carcinogens acrolein, inorganic arsenic, and 2,6-dimethylaniline (2,6-DMA) reach genotoxic concentrations in pet dogs with and without UC. Methods: We first established thresholds for DNA damage from these chemicals using a novel in vitro organoid model. Healthy canine urinary bladder organoids were exposed to acrolein, sodium arsenite, and 2,6-DMA in vitro and we used the alkaline CometChip assay without and with the enzyme Fpg (formamidopyrimidine [fapy]-DNA glycosylase) to measure DNA strand breaks and oxidative DNA damage. Results: For acrolein, we found a genotoxic threshold of 20 uM for combined DNA strand breaks and oxidative DNA damage. These findings suggest potentially genotoxic urinary acrolein exposures in 20% of pet dogs (15 of 74) previously surveyed, with no differences between cases and controls. For inorganic arsenic, we observed genotoxicity at 20 uM in canine organoids; none of 74 pet dogs reached this urinary concentration when assayed at a single time point. For 2,6-DMA, the genotoxic threshold was 0.01 uM for combined DNA strand breaks and oxidative DNA damage. Among dogs previously surveyed, 8% of UC cases (3 of 37) and none of 36 controls reached this threshold (p = 0.07) . Conclusion: Acrolein and 2,6-DMA could reach genotoxic urinary concentrations after household exposures in some pet dogs, and the role of 2,6-DMA in canine bladder cancer risk deserves assessment in a larger sample size. KEYWORDS comet assay, DNA damage, bladder cancer, aromatic amines, arsenic, acrolein Introduction Urothelial carcinoma (UC) is the most common type of bladder cancer in pet dogs, affecting an estimated 20,000 dogs per year ( 1) . Canine UC can lead to hematuria, dysuria, and urinary obstruction, and it typically results in euthanasi

FAQs

What was the study’s main goal?

To determine whether urinary concentrations of acrolein, inorganic arsenic, and 2,6-DMA reach genotoxic levels in pet dogs with and without urothelial carcinoma by using a canine urinary bladder organoid model.

How were genotoxicity levels measured in the study?

Healthy canine urinary bladder organoids were exposed in vitro to each chemical, and genotoxicity was assessed using the alkaline CometChip assay, including measurements with and without the enzyme Fpg to evaluate oxidative DNA damage.

What genotoxicity thresholds did the researchers find?

The study identified a genotoxic threshold of 20 uM for acrolein and 20 uM for inorganic arsenic in canine organoids, and a much lower threshold of 0.01 uM for 2,6-DMA for combined DNA strand breaks and oxidative DNA damage.

Sep 29, 2026

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