Quantification and Characterization of CTCs and Clusters in Pancreatic Cancer by Means of the Hough Transform Algorithm

Circulating Tumor Cells (CTCs) are considered a prognostic marker in pancreatic cancer. In this study we present a new approach for counting CTCs and CTC clusters in patients with pancreatic cancer using the Isoflux(TM) System with the Hough transform algorithm (Hough-Isoflux(TM)). The Hough-Isoflux(TM) approach is based on the counting of an array of pixels with a nucleus and cytokeratin expression excluding the CD45 signal. Total CTCs including free and CTC clusters were evaluated in healthy donor samples mixed with pancreatic cancer cells (PCCs) and in samples from patients with pancreatic ductal adenocarcinoma (PDAC). The Isoflux(TM) System with manual counting was used in a blinded manner by three technicians who used Manual-Isoflux(TM) as a reference. The accuracy of the Hough-Isoflux(TM) approach for detecting PCC based on counted events was 91.00% [84.50, 93.50] with a PCC recovery rate of 80.75 ± 16.41%. A high correlation between the Hough-Isoflux(TM) and Manual-Isoflux(TM) was observed for both free CTCs and for clusters in experimental PCC (R(2) = 0.993 and R(2) = 0.902 respectively). However, the correlation rate was better for free CTCs than for clusters in PDAC patient samples (R(2) = 0.974 and R(2) = 0.790 respectively). In conclusion, the Hough-Isoflux(TM) approach showed high accuracy for the detection of circulating pancreatic cancer cells. A better correlation rate was observed between Hough-Isoflux(TM) approach and with the Manual-Isoflux(TM) for isolated CTCs than for clusters in PDAC patient samples.