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Clinical Trial
. 2020 Feb 21;21(4):1475.
doi: 10.3390/ijms21041475.

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

Vera Cappelletti  1 , Elena Verzoni  2 , Raffaele Ratta  2 , Marta Vismara  1 , Marco Silvestri  1 , Rosanna Montone  3 , Patrizia Miodini  1 , Carolina Reduzzi  1 , Melanie Claps  2 , Pierangela Sepe  2 , Maria Grazia Daidone  1 , Giuseppe Procopio  2
Affiliations
Clinical Trial

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

Vera Cappelletti et al. Int J Mol Sci. .

Abstract

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.

Keywords: cancer evolution; circulating tumor cells; copy number alterations; heterogeneity; renal cell cancer; single-cell analysis.

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Conflict of interest statement

C.R. received two travel grants from Menarini Silicon Biosystems for attending the DEPArray User Meeting in Philadelphia, PA Sept 2018 and the 4th ACTC Meeting at Corfù, Greece October 2019. The other authors declare no conflict of interest. The funders had no role in the design of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or in the decision to publish the results.

Figures

Figure 1
Figure 1
Genomic Identification of Significant Targets in Cancer (GISTIC) amplification (left) and deletion (right) plots on single CTCs. The genome is oriented vertically from top to bottom, and the GISTIC q-values at each locus are plotted from left to right on a log scale. The green line represents the significance threshold (q-value = 0.1).
Figure 2
Figure 2
Matrix reporting the top genomic gains (red) and losses (blue) in our CTCs. Color codes refer to blood collection timing and CTC phenotype.
Figure 3
Figure 3
Matrix reporting genomic gains (red) and losses (blue) commonly observed in renal cell cancer for 14 CTCs isolated from patient K017. Color codes refer to blood collection timing and CTC phenotype.
Figure 4
Figure 4
Translational oncology (TRONCO) was used to order 14 single CTCs isolated from patient K017 into a clonal hierarchy. The figure represents phylogenies of single CTCs grouped into three distinct clones by considering genomic regions altered in at least 50% of all CTCs. Loss at 9p21.3 (indicated by blue color) was observed across all the three clones.
See this image and copyright information in PMC

References

    1. Flanigan R.C., Yonover P.M. The role of radical nephrectomy in metastatic renal cell carcinoma. Semin. Urol. Oncol. 2001;19:98–102. - PubMed
    1. Motzer R.J., Hutson T.E., Tomczak P., Michaelson M.D., Bukowski R.M., Oudard S., Negrier S., Szczylik C., Pili R., Bjarnason G.A., et al. Overall survival and updated results for sunitinib compared with interferon alfa in patients with metastatic renal cell carcinoma. J. Clin. Oncol. 2009;27:3584–3590. doi: 10.1200/JCO.2008.20.1293. - DOI - PMC - PubMed
    1. Sternberg C.N., Davis I.D., Mardiak J., Szczylik C., Lee E., Wagstaff J., Barrios C.H., Salman P., Gladkov O.A., Kavina A., et al. Pazopanib in locally advanced or metastatic renal cell carcinoma: Results of a randomized phase III trial. J. Clin. Oncol. 2010;28:1061–1068. doi: 10.1200/JCO.2009.23.9764. - DOI - PubMed
    1. Motzer R.J., Hutson T.E., Cella D., Reeves J., Hawkins R., Guo J., Nathan P., Staehler M., de Souza P., Merchan J.R., et al. Pazopanib versus sunitinib in metastatic renal-cell carcinoma. N. Engl. J. Med. 2013;369:722–731. doi: 10.1056/NEJMoa1303989. - DOI - PubMed
    1. Flanigan R.C., Salmon S.E., Blumenstein B.A., Bearman S.I., Roy V., McGrath P.C., Caton J.R., Jr., Munshi N., Crawford E.D. Nephrectomy followed by interferon alfa-2b compared with interferon alfa-2b alone for metastatic renal-cell cancer. N. Engl. J. Med. 2001;345:1655–1659. doi: 10.1056/NEJMoa003013. - DOI - PubMed

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