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Open Access 01-12-2021 | Research article

Cell-free DNA and circulating tumor cell kinetics in a pre-clinical head and neck Cancer model undergoing radiation therapy

Authors: Nidal Muhanna, Donovan Eu, Harley H. L. Chan, Catriona Douglas, Jason L. Townson, Marco A. Di Grappa, Reza M. Mohamadi, Shana O. Kelley, Scott V. Bratman, Jonathan C. Irish

Published in: BMC Cancer | Issue 1/2021

Abstract

Background

Monitoring circulating tumor DNA (ctDNA) and circulating tumor cells (CTCs), known as liquid biopsies, continue to be developed as diagnostic and prognostic markers for a wide variety of cancer indications, mainly due to their minimally invasive nature and ability to offer a wide range of phenotypic and genetic information. While liquid biopsies maintain significant promising benefits, there is still limited information regarding the kinetics of ctDNA and CTCs following radiation therapy which remains a vital treatment modality in head and neck cancers. This study aims to describe the kinetics of ctDNA and CTCs following radiation exposure in a preclinical rabbit model with VX2 induced buccal carcinoma.

Methods

Seven rabbits were inoculated with VX2 cells in the buccal mucosa and subjected to radiation. At selected time points, blood sampling was performed to monitor differing levels of ctDNA and CTC. Plasma ctDNA was measured with quantitative PCR for papillomavirus E6 while CTCs were quantified using an immunomagnetic nanoparticles within a microfluidic device. Comparisons of CTC detection with EpCAM compared to multiple surface markers (EGFR, HER2 and PSMA) was evaluated and correlated with the tumor size.

Results

Plasma ctDNA reflects the overall tumor burden within the animal model. Analysis of correlations between ctDNA with tumor and lymph node volumes showed a positive correlation (R = 0.452 and R = 0.433 [p < 0.05]), respectively. Over the course of treatment, ctDNA levels declined and quickly becomes undetectable following tumor eradication. While during the course of treatment, ctDNA levels were noted to rise particularly upon initiation of radiation following scheduled treatment breaks. Levels of CTCs were observed to increase 1 week following inoculation of tumor to the primary site. For CTC detection, the use of multiple surface markers showed a greater sensitivity when compared to detection using only EpCAM. Plasma CTC levels remained elevated following radiation therapy which may account for an increased shedding of CTCs following radiation.

Conclusion

This study demonstrates the utility of ctDNA and CTCs detection in response to radiation treatment in a preclinical head and neck model, allowing for better understanding of liquid biopsy applications in both clinical practice and research development.

Jacobson RA, Munding E, Hayden DM, Levy M, Kuzel TM, Pappas SG, Masood A: Evolving Clinical Utility of Liquid Biopsy in Gastrointestinal Cancers. Cancers (Basel) 2019, 11(8), 11, 8, DOI: https://doi.org/10.3390/cancers11081164.

Luo H, Zhao Q, Wei W, Zheng L, Yi S, Li G, Wang W, Sheng H, Pu H, Mo H, Zuo Z., Liu Z., Li C., Xie C., Zeng Z., Li W., Hao X., Liu Y., Cao S., Liu W., Gibson S., Zhang K., Xu G., Xu R.H.: Circulating tumor DNA methylation profiles enable early diagnosis, prognosis prediction, and screening for colorectal cancer. Sci Transl Med 2020, 12(524), 12, 524, DOI: https://doi.org/10.1126/scitranslmed.aax7533.

Peng M, Xie Y, Li X, Qian Y, Tu X, Yao X, et al. Resectable lung lesions malignancy assessment and cancer detection by ultra-deep sequencing of targeted gene mutations in plasma cell-free DNA. J Med Genet. 2019;56(10):647–53. https://doi.org/10.1136/jmedgenet-2018-105825.CrossRefPubMed

Kulasinghe A, Perry C, Kenny L, Warkiani ME, Nelson C, Punyadeera C. PD-L1 expressing circulating tumour cells in head and neck cancers. BMC Cancer. 2017;17(1):333. https://doi.org/10.1186/s12885-017-3316-3.CrossRefPubMedPubMedCentral

Toss A, Mu Z, Fernandez S, Cristofanilli M. CTC enumeration and characterization: moving toward personalized medicine. Ann Transl Med. 2014;2(11):108. https://doi.org/10.3978/j.issn.2305-5839.2014.09.06.CrossRefPubMedPubMedCentral

Mazel M, Jacot W, Pantel K, Bartkowiak K, Topart D, Cayrefourcq L, et al. Frequent expression of PD-L1 on circulating breast cancer cells. Mol Oncol. 2015;9(9):1773–82. https://doi.org/10.1016/j.molonc.2015.05.009.CrossRefPubMedPubMedCentral

Mandel P, Metais P. Nuclear acids in human blood plasma. C R Seances Soc Biol Fil. 1948;142(3–4):241–3.PubMed

Ashworth TR. A case of Cancer in which cells similar to those in the Tumours were seen in the blood after death. Med J Aust. 1869;14:146–7.

Chan KC, Hung EC, Woo JK, Chan PK, Leung SF, Lai FP, et al. Early detection of nasopharyngeal carcinoma by plasma Epstein-Barr virus DNA analysis in a surveillance program. Cancer. 2013;119(10):1838–44. https://doi.org/10.1002/cncr.28001.CrossRefPubMed

10.

Chan KCA, Woo JKS, King A, Zee BCY, Lam WKJ, Chan SL, et al. Analysis of plasma Epstein-Barr virus DNA to screen for nasopharyngeal Cancer. N Engl J Med. 2017;377(6):513–22. https://doi.org/10.1056/NEJMoa1701717.CrossRefPubMed

11.

Wang Y, Springer S, Mulvey CL, Silliman N, Schaefer J, Sausen M, et al. Detection of somatic mutations and HPV in the saliva and plasma of patients with head and neck squamous cell carcinomas. Sci Transl Med. 2015;7(293):293ra104.CrossRef

12.

Merker JD, Oxnard GR, Compton C, Diehn M, Hurley P, Lazar AJ, et al. Circulating tumor DNA analysis in patients with Cancer: American Society of Clinical Oncology and College of American Pathologists Joint Review. J Clin Oncol. 2018;36(16):1631–41. https://doi.org/10.1200/JCO.2017.76.8671.CrossRefPubMed

13.

Aceto N, Toner M, Maheswaran S, Haber DA. En route to metastasis: circulating tumor cell clusters and epithelial-to-mesenchymal transition. Trends Cancer. 2015;1(1):44–52. https://doi.org/10.1016/j.trecan.2015.07.006.CrossRefPubMed

14.

Mittal V. Epithelial mesenchymal transition in tumor metastasis. Annu Rev Pathol. 2018;13(1):395–412. https://doi.org/10.1146/annurev-pathol-020117-043854.CrossRefPubMed

15.

Allard WJ, Matera J, Miller MC, Repollet M, Connelly MC, Rao C, et al. Tumor cells circulate in the peripheral blood of all major carcinomas but not in healthy subjects or patients with nonmalignant diseases. Clin Cancer Res. 2004;10(20):6897–904. https://doi.org/10.1158/1078-0432.CCR-04-0378.CrossRefPubMed

16.

Figg WD 2nd, Reid J. Monitor tumor burden with circulating tumor DNA. Cancer Biol Ther. 2013;14(8):697–8. https://doi.org/10.4161/cbt.25361.CrossRefPubMed

17.

Ma B, Hui EP, King A, Leung SF, Kam MK, Mo F, et al. Prospective evaluation of plasma Epstein-Barr virus DNA clearance and fluorodeoxyglucose positron emission scan in assessing early response to chemotherapy in patients with advanced or recurrent nasopharyngeal carcinoma. Br J Cancer. 2018;118(8):1051–5. https://doi.org/10.1038/s41416-018-0026-9.CrossRefPubMedPubMedCentral

18.

Muhanna N, Di Grappa MA, Chan HHL, Khan T, Jin CS, Zheng Y, et al. Cell-free DNA kinetics in a pre-clinical model of head and neck Cancer. Sci Rep. 2017;7(1):16723. https://doi.org/10.1038/s41598-017-17079-6.CrossRefPubMedPubMedCentral

19.

Muhanna N, Mepham A, Mohamadi RM, Chan H, Khan T, Akens M, et al. Nanoparticle-based sorting of circulating tumor cells by epithelial antigen expression during disease progression in an animal model. Nanomedicine. 2015;11(7):1613–20. https://doi.org/10.1016/j.nano.2015.04.017.CrossRefPubMed

20.

Chaudhuri AA, Binkley MS, Osmundson EC, Alizadeh AA, Diehn M. Predicting radiotherapy responses and treatment outcomes through analysis of circulating tumor DNA. Semin Radiat Oncol. 2015;25(4):305–12. https://doi.org/10.1016/j.semradonc.2015.05.001.CrossRefPubMedPubMedCentral

21.

Khoja L, Backen A, Sloane R, Menasce L, Ryder D, Krebs M, et al. A pilot study to explore circulating tumour cells in pancreatic cancer as a novel biomarker. Br J Cancer. 2012;106(3):508–16. https://doi.org/10.1038/bjc.2011.545.CrossRefPubMed

22.

Morris KL, Tugwood JD, Khoja L, Lancashire M, Sloane R, Burt D, et al. Circulating biomarkers in hepatocellular carcinoma. Cancer Chemother Pharmacol. 2014;74(2):323–32. https://doi.org/10.1007/s00280-014-2508-7.CrossRefPubMed

23.

Pailler E, Adam J, Barthelemy A, Oulhen M, Auger N, Valent A, et al. Detection of circulating tumor cells harboring a unique ALK rearrangement in ALK-positive non-small-cell lung cancer. J Clin Oncol. 2013;31(18):2273–81. https://doi.org/10.1200/JCO.2012.44.5932.CrossRefPubMed

24.

Kulasinghe A, Kapeleris J, Kimberley R, Mattarollo SR, Thompson EW, Thiery JP, et al. The prognostic significance of circulating tumor cells in head and neck and non-small-cell lung cancer. Cancer Med. 2018;7(12):5910–9. https://doi.org/10.1002/cam4.1832.CrossRefPubMedPubMedCentral

25.

Kulasinghe A, Warkiani ME, Punyadeera C. The isolation and characterization of circulating tumor cells from head and neck Cancer patient blood samples using spiral microfluidic technology. Methods Mol Biol. 2019;2054:129–36. https://doi.org/10.1007/978-1-4939-9769-5_8.CrossRefPubMed

26.

Montero PH, Patel SG. Cancer of the oral cavity. Surg Oncol Clin N Am. 2015;24(3):491–508. https://doi.org/10.1016/j.soc.2015.03.006.CrossRefPubMedPubMedCentral

27.

Choi JJ, Reich CF 3rd, Pisetsky DS. The role of macrophages in the in vitro generation of extracellular DNA from apoptotic and necrotic cells. Immunology. 2005;115(1):55–62. https://doi.org/10.1111/j.1365-2567.2005.02130.x.CrossRefPubMedPubMedCentral

28.

Grisanti S, Almici C, Consoli F, Buglione M, Verardi R, Bolzoni-Villaret A, et al. Circulating tumor cells in patients with recurrent or metastatic head and neck carcinoma: prognostic and predictive significance. PLoS One. 2014;9(8):e103918. https://doi.org/10.1371/journal.pone.0103918.CrossRefPubMedPubMedCentral

29.

Nichols AC, Lowes LE, Szeto CC, Basmaji J, Dhaliwal S, Chapeskie C, Todorovic B, Read N, Venkatesan V, Hammond A et al: Detection of circulating tumor cells in advanced head and neck cancer using the CellSearch system. Head Neck 2012, 34(10):1440–1444, Detection of circulating tumor cells in advanced head and neck cancer using the cellsearch system, DOI: https://doi.org/10.1002/hed.21941.

30.

Lv J, Chen Y, Zhou G, Qi Z, Tan KRL, Wang H, et al. Liquid biopsy tracking during sequential chemo-radiotherapy identifies distinct prognostic phenotypes in nasopharyngeal carcinoma. Nat Commun. 2019;10(1):3941. https://doi.org/10.1038/s41467-019-11853-y.CrossRefPubMedPubMedCentral

31.

Lin JC, Wang WY, Chen KY, Wei YH, Liang WM, Jan JS, et al. Quantification of plasma Epstein-Barr virus DNA in patients with advanced nasopharyngeal carcinoma. N Engl J Med. 2004;350(24):2461–70. https://doi.org/10.1056/NEJMoa032260.CrossRefPubMed

32.

Cao H, Banh A, Kwok S, Shi X, Wu S, Krakow T, et al. Quantitation of human papillomavirus DNA in plasma of oropharyngeal carcinoma patients. Int J Radiat Oncol Biol Phys. 2012;82(3):e351–8. https://doi.org/10.1016/j.ijrobp.2011.05.061.CrossRefPubMed

Title: Cell-free DNA and circulating tumor cell kinetics in a pre-clinical head and neck Cancer model undergoing radiation therapy
Authors: Nidal Muhanna
Donovan Eu
Harley H. L. Chan
Catriona Douglas
Jason L. Townson
Marco A. Di Grappa
Reza M. Mohamadi
Shana O. Kelley
Scott V. Bratman
Jonathan C. Irish
Publication date: 01-12-2021
Publisher: BioMed Central
Published in: BMC Cancer / Issue 1/2021
Electronic ISSN: 1471-2407
DOI: https://doi.org/10.1186/s12885-021-08791-8

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Abstract

Background

Methods

Results

Conclusion

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