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Cell-free circulating tumor DNA in colorectal cancer: a proof of concept with simplified methodology

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Abstract

Background

Cell-free DNA analysis (cfDNA) holds promise for residual disease or tumor burden quantification in colorectal cancer, with reduced costs and diagnostic equipment compared to gold standard-specific tumor DNA (ctDNA) analysis.

Methods

This prospective case–control study included 46 colorectal cancer patients and healthy controls to perform cfDNA quantification by fluorometry using Quantus Fluorometer (Promega, Madison, WI) and using cell-free DNA ScreenTape assay (Agilent) and 4200 TapeStation instrument (Agilent Technologies, Inc., Santa Clara, CA, USA). cfDNA quantification results were correlated with stage, clinical and histopathological features.

Results

33 localized (8 stage I, 12 stage II, and 13 stage III) and 13 advanced colorectal cancer patients were included. No differences in cfDNA quantification by fluorometry were demonstrated depending on stage or histopathological features in localized disease patients. Differences in cfDNA quantification by fluorometry could be demonstrated in patients with advanced disease depending on the presence of liver metastases and synchronous or metachronous metastatic disease. Differences in cfDNA quantification by fluorometry could be demonstrated between advanced colorectal cancer patients and both localized disease patients and healthy controls. Secondary cfDNA analysis by electrophoresis, although showing more specificity to measure ctDNA in cfDNA values, could not improve the capacity to detect differences between analyzed a groups beyond previously achieved with fluorometry.

Conclusion

This exploratory analysis of cfDNA based on fluorometry and electrophoresis methods showed promising results discriminating colorectal cancer and non-cancer patients, as well as different colorectal cancer stages and disease profiles. Further studies are needed to increase our knowledge and to help to overcome barriers to broader implementation and applications.

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Funding

This study was promoted by the Medical Oncology Department of Hospital General Universitario de Elche trough FISABIO (Valencia Biomedical and Health Investigation Promoting Foundation).

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Authors and Affiliations

  1. Medical Oncology Department, Elche University Hospital, Camí Làlmazara s/n, 03203, Elche, Spain

    Javier Bosque, Carlos Guirao, Asia Ferrández, Noelia Suarez, Diana Anguita, María Pamies & Javier Gallego Plazas

  2. Molecular Genetics Unit, Elche University Hospital, 03203, Elche, Spain

    Maria Isabel Castillejo & José Luis Soto

  3. Foundation for the Promotion of Health and Biomedical Research of Valencia Region (FISABIO), FISABIO-Elche Health Department, 03203, Elche, Spain

    Maria Isabel Castillejo & José Luis Soto

  4. Biostatistics Department, FISABIO-Elche Health Department, 03203, Elche, Spain

    Alejandro Moya

Authors
  1. Javier Bosque
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  2. Carlos Guirao
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  3. Asia Ferrández
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  4. Noelia Suarez
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  5. Maria Isabel Castillejo
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  6. Diana Anguita
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  7. María Pamies
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  8. Alejandro Moya
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  9. José Luis Soto
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  10. Javier Gallego Plazas
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Corresponding author

Correspondence to Javier Gallego Plazas.

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Conflict of interest

All authors certify they do not have potential conflicts of interests related to the content of this investigation and manuscript.

Ethical approval

This study was approved by the local Ethics Committee of the Hospital General Universitario de Elche on October 31st of 2018 with registration number PI 39/2018.

Informed consent

Before study entry, all colorectal cancer patients signed informed consent related to study. Non-cancer patient controls were obtained from de Red Valenciana de Biobancos (RVB), a regional institutional Biobank net. All controls signed an informed consent related to the Biobank net.

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Bosque, J., Guirao, C., Ferrández, A. et al. Cell-free circulating tumor DNA in colorectal cancer: a proof of concept with simplified methodology. Clin Transl Oncol 24, 1924–1931 (2022). https://doi.org/10.1007/s12094-022-02841-8

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  • DOI: https://doi.org/10.1007/s12094-022-02841-8

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