1140 EVALUATION OF URINARY EXOSOMES IN BLADDER CANCER PATIENTS

Abstract

You have accessJournal of UrologyBladder Cancer: Basic Research (III)1 Apr 20131140 EVALUATION OF URINARY EXOSOMES IN BLADDER CANCER PATIENTS Jennifer J. Ahn, Guillermo Salazar, Matthew Seaman, Anna Scott, Piruz Motamedinia, Crystal Castaneda, Wayne Comper, James M. McKiernan, and Leileata Russo Jennifer J. AhnJennifer J. Ahn New York, NY More articles by this author , Guillermo SalazarGuillermo Salazar New York, NY More articles by this author , Matthew SeamanMatthew Seaman New York, NY More articles by this author , Anna ScottAnna Scott New York, NY More articles by this author , Piruz MotamediniaPiruz Motamedinia New York, NY More articles by this author , Crystal CastanedaCrystal Castaneda New York, NY More articles by this author , Wayne ComperWayne Comper New York, NY More articles by this author , James M. McKiernanJames M. McKiernan New York, NY More articles by this author , and Leileata RussoLeileata Russo New York, NY More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2013.02.755AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES Despite significant strides in understanding the molecular pathogenesis of bladder cancer (BCa), diagnosis and surveillance continue to rely upon cystoscopy and cytology. As a result, there has been substantial interest in the use of biomarkers for diagnosis and risk stratification. However, current assays rely on tumor cells to be shed into the urine, or examine proteins that may not be specific for cancer. We have previously shown that urinary exosomes and microvesicles (collectively referred to as exosomes) are a consistent and reproducible means of sampling high integrity genetic material, but this has yet to be investigated in the context of BCa. Here, we examined the feasibility of evaluating the urinary exosomes of bladder cancer patients. METHODS Following Columbia University Medical Center IRB approval, voided urine samples were collected from 12 patients. BCa status at time of collection, as determined by cystoscopy and biopsy, was high-grade (HG) tumor (n = 5), low-grade (LG) tumor (n = 2), or no evidence of disease (NED) (n = 5). Samples were stored at 4°C and 0.8 μm filtration was used to remove whole cells and debris. Urinary exosomal RNA (exoRNA) was isolated using an in-house technique (Exosome Diagnostics, NY) and analyzed using RT-qPCR for 18 genes previously described as being differentially expressed in BCa. To compare expression levels, genes were standardized to 18S and relative quantitation (RQ) was calculated using DataAssist software (v.3.0). RESULTS Of the 18 genes analyzed, ICAM1 showed significantly higher expression in patients with HG BCa, as compared to NED patients (RQ 37.0, p = 0.0084). A similar trend was seen when comparing patients with any BCa (HG or LG) to NED patients (RQ 14.2, p = 0.051). UPK2 expression levels were not significantly different between the groups HG versus NED (RQ 1.6, p = 0.958) or LG versus NED (RQ 1.0, p = 0.975). CONCLUSIONS We successfully isolated exosomal RNA from the voided urine of patients with concurrent bladder cancer, as well as those with no evidence of disease. Additionally, significant upregulation of ICAM1 was seen in patients with HG disease. Ultimately, this technique shows promise as a non-invasive method of assessing gene expression in bladder cancer, independent of examining urothelial cells. © 2013 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 189Issue 4SApril 2013Page: e465-e466 Advertisement Copyright & Permissions© 2013 by American Urological Association Education and Research, Inc.MetricsAuthor Information Jennifer J. Ahn New York, NY More articles by this author Guillermo Salazar New York, NY More articles by this author Matthew Seaman New York, NY More articles by this author Anna Scott New York, NY More articles by this author Piruz Motamedinia New York, NY More articles by this author Crystal Castaneda New York, NY More articles by this author Wayne Comper New York, NY More articles by this author James M. McKiernan New York, NY More articles by this author Leileata Russo New York, NY More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...