MP39-13 DEVELOPMENT OF A NOVEL METHOD FOR DETECTING RENAL CELL CARCINOMA CIRCULATING TUMOR CELLS

Abstract

You have accessJournal of UrologyKidney Cancer: Basic Research I1 Apr 2015MP39-13 DEVELOPMENT OF A NOVEL METHOD FOR DETECTING RENAL CELL CARCINOMA CIRCULATING TUMOR CELLS Michael Gorin, Mark Ball, Darren Davis, Phillip Pierorazio, Hans Hammers, Kenneth Pienta, and Mohamad Allaf Michael GorinMichael Gorin More articles by this author , Mark BallMark Ball More articles by this author , Darren DavisDarren Davis More articles by this author , Phillip PierorazioPhillip Pierorazio More articles by this author , Hans HammersHans Hammers More articles by this author , Kenneth PientaKenneth Pienta More articles by this author , and Mohamad AllafMohamad Allaf More articles by this author View All Author Informationhttps://doi.org/10.1016/j.juro.2015.02.763AboutPDF ToolsAdd to favoritesDownload CitationsTrack CitationsPermissionsReprints ShareFacebookTwitterLinked InEmail INTRODUCTION AND OBJECTIVES The isolation of circulating tumor cells (CTCs) from patients with renal cell carcinoma (RCC) has been met with limited success. This is due to the fact that most available CTC isolation technologies rely on the positive selection of cells using the surface protein EpCAM, an epithelial marker which is expressed in a minority of RCCs. ApoStream (ApoCell Inc., Houston, TX) is a novel technology which utilizes dielectrophoresis and microfluidics for the antibody-free isolation of CTCs. In this study, we developed a novel method for detecting RCC CTCs using the ApoStream platform and fluorescence in situ hybridization (FISH) for loss of the VHL gene. This assay was then tested in three patients with metastatic clear cell RCC. METHODS The optimal operating frequency for enrichment of RCC CTCs was determined using fluorescently labeled 786-O cells spiked in peripheral blood mononuclear cells from healthy donors. In parallel, conditions were optimized for performing FISH for the VHL gene (ZytoLight VHL/centromere 3 probe, ZytoVision GmbH, Germany) on isolated cells. Following successful assay development, 7.5 mL of blood from each of three patients with metastatic clear cell RCC was enriched for CTCs using the ApoStream instrument. From each sample, 1400 isolated mononuclear cells were evaluated with the optimized FISH assay. A CTC was defined as any mononuclear cell demonstrating net loss of the VHL gene. RESULTS Five operating frequencies (25, 45, 65, 85, 105 Hz) were evaluated for the optimal isolation of 786-O cells from peripheral blood mononuclear cells. Among the tested frequencies, 85 Hz demonstrated the highest mean rate of cell recovery (65%). In clinical testing, two of three (66.7%) patients had detectable CTCs (3 and 5 CTCs each). Figure 1 includes representative images of cells isolated from these two patients. CONCLUSIONS Antibody-independent isolation with dielectrophoresis and subsequent FISH for the VHL gene is a promising novel method for CTC detection in patients with metastatic RCC. Future work aims to validate this assay in larger patient cohort. © 2015 by American Urological Association Education and Research, Inc.FiguresReferencesRelatedDetails Volume 193Issue 4SApril 2015Page: e459 Advertisement Copyright & Permissions© 2015 by American Urological Association Education and Research, Inc.MetricsAuthor Information Michael Gorin More articles by this author Mark Ball More articles by this author Darren Davis More articles by this author Phillip Pierorazio More articles by this author Hans Hammers More articles by this author Kenneth Pienta More articles by this author Mohamad Allaf More articles by this author Expand All Advertisement Advertisement PDF downloadLoading ...