Longitudinal cell-free DNA characterization by low-coverage whole-genome sequencing in patients undergoing high-dose radiotherapy

Radiother Oncol. 2024 Aug:197:110364. doi: 10.1016/j.radonc.2024.110364. Epub 2024 Jun 2.

Abstract

Background and purpose: Current radiotherapy guidelines rely heavily on imaging-based monitoring. Liquid biopsy monitoring promises to complement imaging by providing frequent systemic information about the tumor. In particular, cell-free DNA (cfDNA) sequencing offers a tumor-agnostic approach, which lends itself to monitoring heterogeneous cohorts of cancer patients.

Methods: We collected plasma cfDNA from oligometastatic patients (OMD) and head-and-neck cancer patients (SCCHN) at six time points before, during, and after radiotherapy, and compared them to the plasma samples of healthy and polymetastatic volunteers. We performed low-pass (on average 7x) whole-genome sequencing on 93 plasma cfDNA samples and correlated copy number alterations and fragment length distributions to clinical and imaging findings.

Results: We observed copy number alterations in 4/7 polymetastatic cancer patients, 1/7 OMD and 1/7 SCCHN patients, these patients' imaging showed progression following radiotherapy. Using unsupervised learning, we identified cancer-specific fragment length features that showed a strong correlation with copy number-based tumor fraction estimates. In 4/4 HPV-positive SCCHN patient samples, we detected viral DNA that enabled the monitoring of very low tumor fraction samples.

Conclusions: Our results indicate that an elevated tumor fraction is associated with tumor aggressiveness and systemic tumor spread. This information may be used to adapt treatment strategies. Further, we show that by detecting specific sequences such as viral DNA, the sensitivity of detecting cancer from cell-free DNA sequencing data can be greatly increased.

Keywords: Cell-free DNA; Copy number alteration; Fragmentomics; Headand-neck cancer; Human papillomavirus; Liquid biopsy; Oligometastasis; Polymetastasis; Radiotherapy.

MeSH terms

  • Adult
  • Aged
  • Cell-Free Nucleic Acids* / blood
  • DNA Copy Number Variations
  • Female
  • Head and Neck Neoplasms* / blood
  • Head and Neck Neoplasms* / genetics
  • Head and Neck Neoplasms* / pathology
  • Head and Neck Neoplasms* / radiotherapy
  • Humans
  • Male
  • Middle Aged
  • Radiotherapy Dosage
  • Squamous Cell Carcinoma of Head and Neck / blood
  • Squamous Cell Carcinoma of Head and Neck / genetics
  • Squamous Cell Carcinoma of Head and Neck / radiotherapy
  • Whole Genome Sequencing*

Substances

  • Cell-Free Nucleic Acids