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Genetics and Genomics

Circulating tumour cells and cell-free DNA as a prognostic factor in metastatic colorectal cancer: the OMITERC prospective study

British Journal of Cancer volume 125, pages 94–100 (2021)Cite this article

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Abstract

Background

Within the OMITERC prospective study (OMIcs application from solid to liquid biopsy for a personalised ThERapy of Cancer), we explored the prognostic role of liquid biopsy encompassing cell-free DNA (cfDNA) and circulating tumour cells (CTCs) in KRAS mutated metastatic colorectal cancer (mCRC).

Methods

We defined a workflow including pre-analytical and analytical procedures collecting blood before therapy and every 3 months until disease progression (PD). CTCs were counted by CellSearch® and isolated by DEPArray™. NGS sequencing of CTCs and cfDNA was performed using a panel of cancer/CRC related genes respectively.

Results

KRAS mutational status was mostly concordant between tumour tissues and liquid biopsy. The percentage of cfDNA samples with mutations in CRC driver genes was in line with literature. In longitudinal monitoring circulating biomarkers anticipated or overlapped conventional diagnostic tools in predicting PD. The presence of CTCs at baseline was confirmed a negative prognostic marker.

Conclusions

Cell-free DNA and CTCs are readily available candidates for clinical application in mCRC. While CTCs demonstrated a prognostic significance at baseline, cfDNA was confirmed an easily accessible material for monitoring the mutational status of the tumour over time. Moreover, in the longitudinal study, the two markers emerged as complementary in assessing disease progression.

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Fig. 1: Kaplan–Meier estimates of probabilities of progression-free survival (PFS) and overall survival (OS) of mCRC patients presenting (CTC POS) or not (CTC NEG) CTCs at baseline (time 0).
Fig. 2: Longitudinal disease monitoring in two mCRC patients by liquid biopsy evaluation.

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References

  1. Zavridou, M., Mastoraki, S., Strati, A., Tzanikou, E., Chimonidou, M. & Lianidou, E. Evaluation of preanalytical conditions and implementation of quality control steps for reliable gene expression and DNA methylation analyses in liquid biopsies. Clin. Chem. 64, 1522–1533 (2018).

    Article  CAS  Google Scholar 

  2. Alix-Panabières, C. & Pantel, K. Clinical applications of circulating tumor cells and circulating tumor DNA as liquid biopsy. Cancer Discov. 6, 479–491 (2016).

    Article  Google Scholar 

  3. Neumann, M. H. D., Bender, S., Krahn, T. & Schlange, T. ctDNA and CTCs in liquid biopsy—current status and where we need to progress. Comput. Struct. Biotechnol. J. 16, 190–195 (2018).

    Article  Google Scholar 

  4. Salvianti, F., Rotunno, G., Galardi, F., De Luca, F., Pestrin, M., Vannucchi, A. M. et al. Feasibility of a workflow for the molecular characterization of single cells by next generation sequencing. Biomol. Detect. Quantif. 5, 23–29 (2015).

    Article  CAS  Google Scholar 

  5. De Luca, F., Rotunno, G., Salvianti, F., Galardi, F., Pestrin, M., Gabellini, S. et al. Mutational analysis of single circulating tumor cells by next generation sequencing in metastatic breast cancer. Oncotarget 7, 26107–26109 (2016).

    Article  Google Scholar 

  6. Molnár, B., Galamb, O., Kalmár, A., Barták, B. K., Nagy, Z. B., Tóth, K. et al. Circulating cell-free nucleic acids as biomarkers in colorectal cancer screening and diagnosis—an update. Expert. Rev. Mol. Diagn. 19, 477–498 (2019).

    Article  Google Scholar 

  7. Cohen, S. J., Punt, C. J., Iannotti, N., Saidman, B. H., Sabbath, K. D., Gabrail, N. Y. et al. Relationship of circulating tumor cells to tumor response, progression-free survival, and overall survival in patients with metastatic colorectal cancer. J. Clin. Oncol. 26, 3213–3221 (2008).

    Article  Google Scholar 

  8. Tol, J., Koopman, M., Miller, M. C., Tibbe, A., Cats, A., Creemers, G. J. et al. Circulating tumour cells early predict progression-free and overall survival in advanced colorectal cancer patients treated with chemotherapy and targeted agents. Ann. Oncol. 21, 1006–1012 (2010).

    Article  CAS  Google Scholar 

  9. Bidard, F. C., Ferrand, F. R., Huguet, F., Hammel, P., Louvet, C., Malka, D. et al. Disseminated and circulating tumor cells in gastrointestinal oncology. Crit. Rev. Oncol. Hematol. 82, 103–115 (2012).

    Article  CAS  Google Scholar 

  10. Raimondi, C., Nicolazzo, C., Gradilone, A., Giannini, G., De Falco, E., Chimenti, I. et al. Circulating tumor cells: exploring intratumor heterogeneity of colorectal cancer. Cancer Biol. Ther. 15, 496–503 (2014).

    Article  CAS  Google Scholar 

  11. Salvianti, F., Pazzagli, M. & Pinzani, P. Single circulating tumor cell sequencing as an advanced tool in cancer management. Expert. Rev. Mol. Diagn. 16, 51–63 (2016).

    Article  CAS  Google Scholar 

  12. Salvianti, F. & Pinzani, P. The diagnostic potential of mutation detection from single circulating tumor cells in cancer patients. Expert. Rev. Mol. Diagn. 17, 975–981 (2017).

    Article  CAS  Google Scholar 

  13. Fehm, T. N., Meier-Stiegen, F., Driemel, C., Jäger, B., Reinhardt, F., Naskou, J. et al. Diagnostic leukapheresis for CTC analysis in breast cancer patients: CTC frequency, clinical experiences and recommendations for standardized reporting. Cytom. A. 93, 1213–1219 (2018).

    Article  Google Scholar 

  14. Kin, C., Kidess, E., Poultsides, G. A., Visser, B. C. & Jeffrey, S. S. Colorectal cancer diagnostics: biomarkers, cell-free DNA, circulating tumor cells and defining heterogeneous populations by single-cell analysis. Expert. Rev. Mol. Diagn. 13, 581–599 (2013).

    Article  CAS  Google Scholar 

  15. Huang, D., Sun, W., Zhou, Y., Li, P., Chen, F., Chen, H. et al. Mutations of key driver genes in colorectal cancer progression and metastasis. Cancer Metastasis Rev. 37, 173–187 (2018).

    Article  CAS  Google Scholar 

  16. Boussios, S., Ozturk, M. A., Moschetta, M., Karathanasi, A., Zakynthinakis-Kyriakou, N., Katsanos, K. H. et al. The developing story of predictive biomarkers in colorectal cancer. J. Pers. Med. 9, E12 (2019).

    Article  Google Scholar 

  17. Rosty, C., Young, J. P., Walsh, M. D., Clendenning, M., Sanderson, K., Walters, R. J. et al. PIK3CA activating mutation in colorectal carcinoma: associations with molecular features and survival. PLoS ONE 8, e65479 (2013).

    Article  CAS  Google Scholar 

  18. McCarthy, A. J. & Chetty, R. Smad4/DPC4. J. Clin. Pathol. 71, 661–664 (2018).

    Article  Google Scholar 

  19. Korphaisarn, K., Morris, V. K., Overman, M. J., Fogelman, D. R., Kee, B. K., Raghav, K. P. S. et al. FBXW7 missense mutation: a novel negative prognostic factor in metastatic colorectal adenocarcinoma. Oncotarget 8, 39268–39279 (2017).

    Article  Google Scholar 

  20. Chang, C. C., Lin, H. H., Lin, J. K., Lin, C. C., Lan, Y. T., Wang, H. S. et al. FBXW7 mutation analysis and its correlation with clinicopathological features and prognosis in colorectal cancer patients. Int. J. Biol. Markers 30, e88–e95 (2015).

    Article  CAS  Google Scholar 

  21. Jardim, D. L., Wheler, J. J., Hess, K., Tsimberidou, A. M., Zinner, R., Janku, F. et al. FBXW7 mutations in patients with advanced cancers: clinical and molecular characteristics and outcomes with mTOR inhibitors. PLoS ONE 9, e89388 (2014).

    Article  Google Scholar 

Download references

Acknowledgements

We wish to thank the OMITERC consortium and the Fondazione Sandro Pitigliani per la lotta contro i tumori ONLUS.

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Author notes

  1. These authors contributed equally: Pamela Pinzani, Lorenzo Antonuzzo

Authors and Affiliations

  1. Department of Experimental and Clinical Biomedical Sciences “Mario Serio”, University of Florence, Florence, Italy

    Francesca Salvianti, Stefania Gelmini, Irene Mancini, Mario Pazzagli & Pamela Pinzani

  2. Clinical Oncology Unit, Careggi University Hospital, Florence, Italy

    Serena Pillozzi, Elisa Giommoni, Marco Brugia, Francesco Di Costanzo & Lorenzo Antonuzzo

  3. Sandro Pitigliani Medical Oncology Department, Hospital of Prato, Istituto Toscano Tumori, Prato, Italy

    Francesca Galardi & Francesca De Luca

  4. Histopathology and Molecular Diagnostics Unit, Careggi University Hospital, Florence, Italy

    Francesca Castiglione

  5. Department of Experimental and Clinical Medicine, University of Florence, Florence, Italy

    Luca Messerini & Lorenzo Antonuzzo

Authors
  1. Francesca Salvianti
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  2. Stefania Gelmini
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Contributions

F.S.: single CTC isolation, mutational analysis of CTCs and cfDNA, data analysis and manuscript writing; S.G.: management of clinical samples, data analysis and manuscript writing; I.M.: NGS data analysis; F.G. and F.D.L.: CTC counting by CellSearch; S.P.: clinical data management and critical revision of the manuscript; E.G. and M.B.: patients’ enrolment, clinical data collection and analysis; F.C. and L.M. mutational analysis of tumour tissues; M.P. and F.D.C.: conceptualisation, funding acquisition and critical revision of manuscript; P.P.: study design, project supervision and manuscript writing; L.A.: patients’ enrolment, clinical data management and study design.

Corresponding author

Correspondence to Pamela Pinzani.

Ethics declarations

Ethics approval and consent to participate

The study was approved by the Local Ethical Committee (Comitato Etico di Area Vasta Centro AOU Careggi, Firenze, Italy), reference number CEAVC BIO 16.028_10033_bio. Written informed consent to participate was obtained from all patients. The study was performed in accordance with the Declaration of Helsinki.

Consent to publish

Not applicable.

Data availability

The datasets analysed during the current study are available from the corresponding author on reasonable request.

Competing interests

The authors declare no competing interests.

Funding information

This research was funded by Regione Toscana FAS 2007-2013, call FAS salute 2014, OMITERC project, grant 4421.02102014.072000037.

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Salvianti, F., Gelmini, S., Mancini, I. et al. Circulating tumour cells and cell-free DNA as a prognostic factor in metastatic colorectal cancer: the OMITERC prospective study. Br J Cancer 125, 94–100 (2021). https://doi.org/10.1038/s41416-021-01399-6

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