Non-invasive detection of mutagenic exposure to aristolochic acid using ultra-deep sequencing of blood and urine samples
- 1Kirk Kerkorian School of Medicine, University of Nevada, Las Vegas; Nevada Institute of Personalized Medicine, University of Nevada, Las Vegas
Abstract
Aristolochic acids (AAs) are potent nephrotoxins and carcinogens found in certain herbal remedies, driving a high burden of urothelial and liver malignancies globally. Currently, at a genomic level, AA exposure is detected retrospectively via DNA sequencing of clonal tumors, which limits opportunities to detect the mutational imprints of the exposure before tumor formation, when risk stratification may be possible. In a cohort of 12 urothelial carcinoma of the urinary tract patients, we profiled matched tumor and normal tissues alongside peripheral blood, urine sediments, and sorted leukocyte fractions. Rather than seeking tumor-derived clonal mutations, we instead interrogate non-clonal mutations associated with AA exposure. Here, we demonstrate that AA-induced mutagenesis can be non-invasively identified using ultra-deep duplex sequencing (DS), with the canonical AA signature, SBS22a, recovered in the blood and urine from patients harboring AA-positive tumors, but absent in AA-negative tumors. Single-cell WGS and DS of fractionated immune cells localized the SBS22a signal to the adaptive lymphoid compartment, identifying these long-lived cells as a systemic reservoir of mutational history. These findings show that a urothelial carcinogen leaves a durable, cell-type-resolved record in long-lived circulating lymphocytes, and establish a proof-of-concept framework for non-invasive carcinogen exposure surveillance.
Related
- Case Report: Necrotizing fasciitis caused by Vibrio vulnificus without recalled seafood or seawater exposure diagnosed by metagenomic next-generation sequencing
- Haplotype-based non-invasive prenatal testing of F8 complex rearrangement via targeted capture sequencing of maternal plasma.
- Exploring vertical genomic heterogeneity and chemotherapy response using targeted genomic sequencing in muscle-invasive bladder cancer
- Src Homology Phosphatase 2 Disruption in Podocytes Attenuates Markers of Aristolochic Acid 1-Induced Renal Injury, Inflammation, and Fibrosis in Mice
- Urinary dual-site proenkephalin (PENK) methylation testing for detection of high-grade or invasive recurrence during surveillance of non-muscle-invasive bladder cancer
BibTeX
@misc{nguyen2026noninvasive,
title = {{Non-invasive detection of mutagenic exposure to aristolochic acid using ultra-deep sequencing of blood and urine samples}},
author = {P. T. Nguyen and A. Boot and P.-H. Lin and S. Sharma and W. Yu and M. Liu and N. Jiang and E. Schmidt and C. C. Valentine and V. Chew and J. J. Salk and F. Y. Lo and J. S. T. Pang and S. G. Rozen and M. Zhivagui},
howpublished = {medRxiv},
year = {2026},
month = oct,
doi = {10.64898/2026.10.02.26363145},
url = {https://www.medrxiv.org/content/10.64898/2026.10.02.26363145v1},
note = {CC BY},
}
RIS
TY - UNPB
TI - Non-invasive detection of mutagenic exposure to aristolochic acid using ultra-deep sequencing of blood and urine samples
AU - P. T. Nguyen
AU - A. Boot
AU - P.-H. Lin
AU - S. Sharma
AU - W. Yu
AU - M. Liu
AU - N. Jiang
AU - E. Schmidt
AU - C. C. Valentine
AU - V. Chew
AU - J. J. Salk
AU - F. Y. Lo
AU - J. S. T. Pang
AU - S. G. Rozen
AU - M. Zhivagui
PY - 2026
DA - 2026/10/02
PB - medRxiv
DO - 10.64898/2026.10.02.26363145
UR - https://www.medrxiv.org/content/10.64898/2026.10.02.26363145v1
AB - Aristolochic acids (AAs) are potent nephrotoxins and carcinogens found in certain herbal remedies, driving a high burden of urothelial and liver malignancies globally. Currently, at a genomic level, AA exposure is detected retrospectively via DNA sequencing of clonal tumors, which limits opportunities to detect the mutational imprints of the exposure before tumor formation, when risk stratification may be possible. In a cohort of 12 urothelial carcinoma of the urinary tract patients, we profiled matched tumor and normal tissues alongside peripheral blood, urine sediments, and sorted leukocyte fractions. Rather than seeking tumor-derived clonal mutations, we instead interrogate non-clonal mutations associated with AA exposure. Here, we demonstrate that AA-induced mutagenesis can be non-invasively identified using ultra-deep duplex sequencing (DS), with the canonical AA signature, SBS22a, recovered in the blood and urine from patients harboring AA-positive tumors, but absent in AA-negative tumors. Single-cell WGS and DS of fractionated immune cells localized the SBS22a signal to the adaptive lymphoid compartment, identifying these long-lived cells as a systemic reservoir of mutational history. These findings show that a urothelial carcinogen leaves a durable, cell-type-resolved record in long-lived circulating lymphocytes, and establish a proof-of-concept framework for non-invasive carcinogen exposure surveillance.
ER -