Cytologically-targeted next generation sequencing: A synergy for diagnosing urothelial carcinoma

Abstract: Introduction Cytology and cystoscopy are used to detect urothelial cancer (UC), but together they still fail to detect some UC cases and are not suitable for screening asymptomatic individuals. Mutations are present in more than 98% of UC, mutations have therapeutic significance, and they can be detected by next generation sequencing (NGS) in urine samples. We review the role of NGS in UC detection. Materials and Methods Comprehensive literature review on UC genetics, economics of NGS, and previous reports of UC detection by NGS. Results The raw costs of NGS have decreased to about 14,000 base pairs per penny, making it appear economically feasible to use NGS widely. Reported NGS assays fall short of predicted sensitivity. Decreased sensitivity is attributed to a low frequency of mutant alleles in many urine samples. Attempts to increase the percentage of mutant alleles, by using cell-free urinary DNA, or by using cell sorting and microfluidics, have been unsuccessful or remain unproven. However, cytologic examination can immediately enable NGS: Urine cytologies with sufficient proportions of abnormal cells could be directly triaged to NGS with high sensitivity for UC detection. For cases with a low proportion of abnormal cells, cytologically-targeted micro-dissection of cells for NGS should maintain sensitivity and decrease sequencing costs. Cytologically-targeted urothelial cells for NGS could allow a screening test for low grade UC. Conclusion Cytology is immediately poised to allow NGS to improve the diagnosis of UC, allowing NGS to be an ancillary test for atypical cytologies, and potentially allowing a screening test for low-grade UC.