MP66-14 DEVELOPMENT & VALIDATION OF A METHOD FOR DETECTION OF CIRCULATING TUMOUR DNA IN MUSCLE-INVASIVE BLADDER CANCER

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research & Pathophysiology III (MP66)1 Sep 2021MP66-14 DEVELOPMENT & VALIDATION OF A METHOD FOR DETECTION OF CIRCULATING TUMOUR DNA IN MUSCLE-INVASIVE BLADDER CANCER Ruchira Nandurkar, Pavel Sluka, Hady Wardan, Ian Davis, and Shomik Sengupta Ruchira NandurkarRuchira Nandurkar More articles by this author , Pavel SlukaPavel Sluka More articles by this author , Hady WardanHady Wardan More articles by this author , Ian DavisIan Davis More articles by this author , and Shomik SenguptaShomik Sengupta More articles by this author View All Author Informationhttps://doi.org/10.1097/JU.0000000000002106.14AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookLinked InTwitterEmail Abstract INTRODUCTION AND OBJECTIVE: Survival from muscle-invasive Bladder Cancer (MIBC) remains variable despite optimal therapy. Current clinical and pathological variables are limited in their prognostic and predictive utility. Identifying the presence and amount of circulating tumour DNA (ctDNA) in the bloodstream is of great interest as an additional tool. Currently, this relies on next-generation sequencing of tumour samples, which is expensive and can take a long time for results, thus precluding responsive clinical decision-making. This study aimed to develop a quick, sensitive, inexpensive and reliable method for detection of ctDNA in MIBC using droplet digital PCR (ddPCR). METHODS: We selected a combination of 10 mutations found in MIBC that were not only common, but when sequentially added to a gene panel, each would increase the panel’s capacity to detect a wider cohort of MIBC patients, thereby maximising the detection rate whilst maintaining assay feasibility. Mutations were selected based upon data published by The Cancer Genome Atlas (TCGA). Our 10-gene panel was tested against archival MIBC specimens. DNA extracted from archival paraffin-embedded tissue blocks was run against the gene panel using ddPCR to assess for presence and fractional abundance of the mutation in each sample. RESULTS: To date, 66 MIBC samples have been processed, of which 65 possessed at least one of the mutations within our gene panel, conferring a 98.5% detection rate, with a median of five panel mutations detected per sample (range 0-9). Our use of ddPCR may be accountable for our higher detection rate compared to the 40% predicted from the TCGA, which used Next Generation Sequencing technology. Further analyses are being undertaken to compare samples before and after neo-adjuvant chemotherapy. CONCLUSIONS: This technique utilising a 10 gene panel appears to be sensitive enough to detect mutations in the majority of patients with MIBC. Prospective validation is underway to determine the potential clinical utility of this technique. Source of Funding: Robert Bulley Research and Innovation Grant, Eastern Health Foundation © 2021 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 206Issue Supplement 3September 2021Page: e1134-e1135 Advertisement Copyright & Permissions© 2021 by American Urological Association Education and Research, Inc.MetricsAuthor Information Ruchira Nandurkar More articles by this author Pavel Sluka More articles by this author Hady Wardan More articles by this author Ian Davis More articles by this author Shomik Sengupta More articles by this author Expand All Advertisement Loading ...