TERT promoter mutated circulating tumor DNA as a biomarker for prognosis in hepatocellular carcinoma

Abstract

Background and aims

Plasma circulating tumor DNA (ctDNA) with tumor-specific mutations is an attractive biomarker. The telomerase reverse transcriptase (TERT) C228T promoter mutation is the most prevalent tumor-associated mutation in hepatocellular carcinoma (HCC). We evaluated the presence and prognostic value of the TERT C228T mutation in plasma and tissue in a Danish HCC cohort.

Methods

We analyzed ctDNA from 95 HCC patients and 45 liver cirrhotic patients without HCC for the TERT mutation using droplet digital polymerase chain reaction. We also analyzed DNA from the corresponding primary tumor tissues in 34 HCC patients.

Results

The plasma TERT C228T mutation was detected in 42/95 HCC patients (44%) but in none of the non-HCC patients. The TERT mutation was detected in 23/34 tumor samples (68%). The TERT mutation was associated with increased mortality when detected in plasma (adjusted HR 2.16 (1.20–3.88), p = .010) but not in tumor tissue (adjusted HR 1.11 (0.35–3.56), p = .860). There was a positive correlation between the presence of the TERT mutation in plasma and an advanced TNM stage (p < .0001) and vascular invasion (p = .005). Analysis of the TERT mutation in plasma and tumor DNA from the same patient was concordant in 21/34 samples (62%; kappa value 0.31, p = .014). Non-concordance was associated with an early TNM stage.

Conclusion

The plasma TERT mutation was detected in 44% of HCC patients and in none of non-HCC cirrhotic patients; and was associated with increased mortality. We propose the TERT C228T mutation in ctDNA as a promising HCC biomarker for prognosis.

Keywords:

• Hepatobiliary-clinical
• hepatocellular carcinoma
• circulating tumor DNA
• droplet digital PCR
• cancer genomics
• oncology-clinical

Acknowledgments

We thank clinical personnel for helping gather samples and laboratory technicians at Department of Hepatology and Gastroenterology, Clinical Biochemistry and Pathology for sample preparation.

Ethical approval

This study was approved by the Central Denmark Region Committee on Biomedical Research Ethics (no. 1-10-72-240-16) and conducted in accordance with the Declaration of Helsinki. All participants gave written informed consent to participate in the study.

Disclosure statement

No potential conflict of interest was reported by the author(s).

Author contribution

SKO, JK, HG, BW and BSS designed the study. SKO authored the first draft of the manuscript and performed the statistical analysis. MSC and BSS developed the assay for TERT analysis and validated the assay. MP evaluated all CT scans. NKA and GEV collected samples from cirrhosis-only patients including interpretation of results and clinical data. SJHD secured data from tissue samples including extraction of DNA and revision of samples. All authors were involved in revising and editing the final manuscript.

Additional information

Funding

Danish Cancer Society, Aarhus University, Health Research Fund of Central Denmark Region, Einar Willumsen Memorial Fund.