American Association for Cancer Research
10780432ccr200401-sup-236661_3_supp_6438200_qdrkdv.pdf (633.4 kB)

Supplementary Figure S2 from Noninvasive Detection of Urothelial Carcinoma by Cost-effective Low-coverage Whole-genome Sequencing from Urine-Exfoliated Cell DNA

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journal contribution
posted on 2023-03-31, 22:12 authored by Shuxiong Zeng, Yidie Ying, Naidong Xing, Baiyun Wang, Ziliang Qian, Zunlin Zhou, Zhensheng Zhang, Weidong Xu, Huiqing Wang, Lihe Dai, Li Gao, Tie Zhou, Jiatao Ji, Chuanliang Xu

Comparison of UroCAD results of urine samples stored for 2 hours and 72 hours.


Shanghai Sailing Program

Discipline development plan in Changhai Hospital

National Natural Science Foundation of China

Science and Technology Commission of Shanghai City

Shanghai Key Laboratory of Cell Engineering

Science and Technology Committee of Shanghai City

National Science and Technology Major Project

Shanghai Clinical Medical Center of Urological Diseases Program

Department of Science & Technology



Urothelial carcinoma is a malignant cancer with frequent chromosomal aberrations. Here, we investigated the application of a cost-effective, low-coverage whole-genome sequencing technology in detecting all chromosomal aberrations. Patients with urothelial carcinomas and nontumor controls were prospectively recruited in clinical trial NCT03998371. Urine-exfoliated cell DNA was analyzed by Illumina HiSeq XTen, followed by genotyping with a customized bioinformatics workflow named Urine Exfoliated Cells Copy Number Aberration Detector (UroCAD). In the discovery phase, urine samples from 126 patients with urothelial carcinomas and 64 nontumor disease samples were analyzed. Frequent chromosome copy-number changes were found in patients with tumor as compared with nontumor controls. A novel diagnosis model, UroCAD, was built by incorporating all the autosomal chromosomal changes. The model reached performance of AUC = 0.92 (95% confidence interval, 89.4%–97.3%). At the optimal cutoff, |Z| ≥ 3.21, the sensitivity, specificity, and accuracy were 82.5%, 96.9%, and 89.0%, respectively. The prediction positivity was found correlated with tumor grade (P = 0.01). In the external validation cohort of 95 participants, the UroCAD assay identified urothelial carcinomas with an overall sensitivity of 80.4%, specificity of 94.9%, and AUC of 0.91. Meanwhile, UroCAD assay outperformed cytology tests with significantly improved sensitivity (80.4% vs. 33.9%; P < 0.001) and comparable specificity (94.9% vs. 100%; P = 0.49). UroCAD could be a robust urothelial carcinoma diagnostic method with improved sensitivity and similar specificity as compared with cytology tests. It may be used as a noninvasive approach for diagnosis and recurrence surveillance in urothelial carcinoma prior to the use of cystoscopy, which would largely reduce the burden on patients.