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. 2023 Feb 21;24(5):4278.
doi: 10.3390/ijms24054278.

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

Francisco José Calero-Castro  1   2 , Sheila Pereira  1   2 , Imán Laga  1   2 , Paula Villanueva  1   2 , Gonzalo Suárez-Artacho  1   2 , Carmen Cepeda-Franco  1   2 , Patricia de la Cruz-Ojeda  2 , Elena Navarro-Villarán  2 , Sandra Dios-Barbeito  1   2 , María José Serrano  3 , Cristóbal Fresno  4 , Javier Padillo-Ruiz  1   2
Affiliations

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

Francisco José Calero-Castro et al. Int J Mol Sci. .

Abstract

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 IsofluxTM System with the Hough transform algorithm (Hough-IsofluxTM). The Hough-IsofluxTM 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 IsofluxTM System with manual counting was used in a blinded manner by three technicians who used Manual-IsofluxTM as a reference. The accuracy of the Hough-IsofluxTM 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-IsofluxTM and Manual-IsofluxTM was observed for both free CTCs and for clusters in experimental PCC (R2 = 0.993 and R2 = 0.902 respectively). However, the correlation rate was better for free CTCs than for clusters in PDAC patient samples (R2 = 0.974 and R2 = 0.790 respectively). In conclusion, the Hough-IsofluxTM approach showed high accuracy for the detection of circulating pancreatic cancer cells. A better correlation rate was observed between Hough-IsofluxTM approach and with the Manual-IsofluxTM for isolated CTCs than for clusters in PDAC patient samples.

Keywords: Hough transform; circulating tumor cell; cluster; pancreatic cancer.

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Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Row one shows the CTC staining characterized by Hoechst 33342+/CK+/CD45-. The fifth image in the first row shows the selection of pixels with CTC staining, and the last photo shows a CTC surrounded by a red circumference counted by the Hough transform. Row two shows the staining of some lymphocytes. Row three shows a free-CTC and a cluster consisting of two cells. Columns five and six show that the Hough transform algorithm did not count the CTCs if the element had no CK staining.
Figure 2
Figure 2
(a). Correlation of total PCCs in control blood samples between control and Manual-IsofluxTM System. (b). Total PCCs in control blood samples between control and Hough-IsofluxTM System. (c). Total PCCs in control blood samples between Manual-IsofluxTM System and Hough-IsofluxTM System.
Figure 3
Figure 3
(a). Cluster measurement in control blood samples: Manual-IsofluxTM System vs. Hough-IsofluxTM System. (b). Clustered-PCC measurement in control blood samples: Manual-IsofluxTM System vs. Hough-IsofluxTM System. (c). Free PCC measurement in control blood samples: Manual-IsofluxTM System vs. Hough-IsofluxTM System.
Figure 4
Figure 4
Correlation plot of CTC/mL in pancreatic cancer patient samples counted with the Manual-IsofluxTM System and with the Hough-IsofluxTM System. (a). Total-CTC correlation. (b). Free-CTC correlation. (c). Cluster correlation. (d). Clustered-CTC correlation.
See this image and copyright information in PMC

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