The Effect of Different Surgical Methods on the Number of Circulating Tumor Cells in the Peripheral Blood of Patients with Renal Cell Carcinoma

e21148 Background: Enumeration of circulating tumor cells (CTCs) in peripheral blood represents a prognostic marker for select solid tumors. Commercially available technologies for CTC detection rely on enrichment of EpCAM-expressing cells. However, not all tumor histologies demonstrate high constitutive EpCAM expression, including renal cell carcinoma (RCC). New techniques for more complex immunophenotypic characterization of CTCs are therefore of great interest. Methods: The AccuCyte system for characterization of rare blood cell populations retains all nucleated cells for analysis. Fluorochrome-conjugated monoclonal antibodies (mAbs) are added to 3 ml peripheral blood samples processed by density separation with a specialized tube and float. Centrifugation deposits all nucleated cells (Buffy coat) onto a surface designed for imaging with an automated fluorescence microscope. Sensitivity for CTC identification was measured by spiking blood from healthy donors with defined numbers of MDA-MD-453 breast tumor cells analyzed with mAbs to EpCAM and CD45. Peripheral blood samples from 4 patients (pts) with metastatic RCC and whose primary tumor had been resected were analyzed with a mAb specific for the RCC-associated antigen carbonic anhydrase IX (CAIX), as well as mAbs for CD10, EGFR and EpCAM. Results: In 13 blood samples from healthy donors, an average of 89% of spiked-in MDA-MD-453 tumor cells defined as EpCAM+/CD45- were detected by AccuCyte analysis with linearity from 0 to 139 cells (slope=0.8899; r2=0.9351) and a sensitivity of approximately 1 CTC per 106 nucleated blood cells. 3 of 4 peripheral blood samples from pts with clear cell RCC contained putative CTCs (range 1-6 cells/ml) defined as CD45-/CAIX+ nucleated cells. 3 CTCs from one patient were also CD10+ or EGFR+. All putative CTCs were EpCAM-. Conclusions: The detection of EpCAM-/CAIX+ cells in RCC pts demonstrates the adaptability of the AccuCyte platform for CTC (or other rare blood cell) identification defined by customized markers. Further development of marker panels for CTC analyses in pts with RCC and other tumor histologies as well as automation of image analysis is ongoing.

Download Full-text

Abstract 5215: Detection of circulating tumor cells in peripheral blood of patients with renal cell carcinoma

10.1158/1538-7445.am2011-5215 ◽

2011 ◽

Author(s):

Steffen Goebel ◽

Karen Bluemke ◽

Udo Bilkenroth ◽

Axel Meye ◽

Susanne Fuessel ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Peripheral Blood ◽

Renal Cell

Download Full-text

Prognostic Role of Circulating Tumor Cells ( CTCS ) in Metastatic Renal Cell Carcinoma: A Large, Multicenter Prospective Trial

The Oncologist ◽

10.1002/onco.13842 ◽

2021 ◽

Author(s):

Umberto Basso ◽

Antonella Facchinetti ◽

Elisabetta Rossi ◽

Marco Maruzzo ◽

Vincenza Conteduca ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Metastatic Renal Cell Carcinoma ◽

Renal Cell ◽

Prospective Trial ◽

Prognostic Role

Download Full-text

Analysis of Single Circulating Tumor Cells in Renal Cell Carcinoma Reveals Phenotypic Heterogeneity and Genomic Alterations Related to Progression

International Journal of Molecular Sciences ◽

10.3390/ijms21041475 ◽

2020 ◽

Vol 21 (4) ◽

pp. 1475

Author(s):

Vera Cappelletti ◽

Elena Verzoni ◽

Raffaele Ratta ◽

Marta Vismara ◽

Marco Silvestri ◽

...

Keyword(s):

Renal Cell Carcinoma ◽

Cell Carcinoma ◽

Tumor Cells ◽

Circulating Tumor Cells ◽

Renal Cell ◽

Progression Free Survival ◽

Response Prediction ◽

Surface Proteins ◽

Cell Recovery ◽

Epithelial Cell Surface

Circulating tumor cells (CTCs) are promising biomarkers for prognosis, therapeutic response prediction, and treatment monitoring in cancer patients. Despite its epithelial origin, renal cell carcinoma (RCC) shows low expression of epithelial markers hindering CTC-enrichment approaches exploiting epithelial cell surface proteins. In 21 blood samples serially collected from 10 patients with metastatic RCC entering the TARIBO trial, we overcame this limitation using the marker-independent Parsortix™ approach for CTC-enrichment coupled with positive and negative selection with the DEPArray™ with single cell recovery and analysis for copy number alterations (CNA) by next generation sequencing NGS. Two CTC subpopulations were identified: epithelial CTC (eCTC) and non-conventional CTC (ncCTC) lacking epithelial and leukocyte markers. With a threshold ≥1CTC/10 mL of blood, the positivity rates were 28% for eCTC, 62% for ncCTCs, and 71% considering both CTC types. In two patients with detectable eCTCs at baseline, progression free survival was less than 5 months. In an index case, hierarchical structure by translational oncology (TRONCO) identified three clones among 14 CTCs collected at progression and at baseline, each containing cells with a 9p21.3loss, a well-known metastasis driving subclonal alteration. CTCs detection in RCC can be increased by marker-independent approaches, and CTC molecular characterization can allow detection of subclonal events possibly related to tumor progression.

Download Full-text