Feasibility Study of Blood Biomarker Identification in Cervical Cancer

This study is Open label, prospective, experimental study of patients with cervical cancer.

The primary aim of this study is to evaluate the feasibility, clinical utility and analytical performance of extracellular vesicles (EVs) and cell-free DNA (cfDNA) as minimally invasive blood-based biomarkers for the early detection, treatment monitoring and post-treatment surveillance of cervical cancer, with the potential to inform future screening and therapeutic strategies.

Cervical cancer continues to impose significant physical, emotional, and financial burdens on women worldwide. Despite advances in prevention and treatment, there remains a critical need for more effective, non-invasive tools to support early detection, monitor treatment response, and guide clinical decision-making. As the field of cervical cancer diagnosis evolves, there is growing interest in the use of blood-based biomarkers, particularly extracellular vesicles (EVs) and cell-free DNA (cfDNA), for improving patient care.

This study aims to evaluate the potential of EVs and cfDNA as minimally invasive biomarkers in the management of cervical cancer. By analysing EVs and cfDNA, we seek to gain deeper insights into their roles in tumour development, progression, and intercellular communication. The anticipated outcomes include the identification of EV- and cfDNA-based signatures that can:

• Facilitate earlier detection of cervical cancer
• Enable real-time monitoring of treatment response
• Predict recurrence and disease progression
• Support the development of personalised therapeutic strategies

By generating preliminary comparative data on the diagnostic accuracy of EVs and cfDNA, this study will provide a critical foundation for future large-scale investigations. It aims to contribute to the development of a novel, blood-based screening and monitoring tool that could enable earlier intervention and improve long-term outcomes.

A reliable, non-invasive test would also serve as a more accessible and acceptable alternative to current cervical screening methods, particularly for women who face cultural, systemic, or personal barriers to participation. This is especially important for under-screened populations, including Aboriginal and Torres Strait Islander women and culturally diverse communities.

Ultimately, this research is expected to advance our understanding of the biological role of EVs and cfDNA in cervical cancer pathogenesis and progression. It supports the development of clinically effective, personalised diagnostic tools that improve outcomes, reduce healthcare costs, and contribute to Australia's national strategy to eliminate cervical cancer as a public health issue while addressing persistent inequities in access to care.

Study Intervention:

This prospective cohort study will enrol 100 patients with stage IA to IVB cervical cancer who are undergoing either primary surgical treatment, radiotherapy (RT) or chemoradiotherapy (CRT). The intervention in this study involves the collection of approximately 40mL of blood at baseline, following the initial diagnosis of cervical cancer (for the analysis of cell-free DNA, extracellular vesicles, and germline DNA). Additional blood samples of the same volume will be collected at two subsequent time points: at 8 weeks following surgery or completion of chemoradiation therapy (CRT), and at 6 months post-baseline. In addition, quality-of-life questionnaires (as detailed in Sections 7) will be completed by participants at the following time points:

• The first timepoint will be at baseline: In patients who have primary surgery, baseline is the date of surgery. In patients who have definitive radiotherapy or chemoradiotherapy, the baseline is the date of first radiation dose. Baseline assessments can be performed up to 30 days prior to the date of surgery or up to 30 days prior to the first radiation dose.
• The second timepoint will occur immediately after completion of radiotherapy, chemoradiotherapy (+ 14 days) or 8 weeks after surgery (± 14 days).
• The third timepoint will be 6 months post baseline (±14 days). Follow up (24 months): Follow up of participant disease status will be recorded at 24 months (± 30 days) by accessing the participant's medical records.

In accordance with current ESMO (European Society for Medical Oncology) guidelines [31], patients diagnosed with early-stage invasive cervical cancer (e.g., FIGO stage IA to IB2, and selected IIA1) typically undergo primary surgical treatment, such as radical hysterectomy or simple hysterectomy. For these patients, the second blood collection will occur at 8 weeks post-surgery (±14 days).

For patients with locally advanced cervical cancer (LACC), in whom primary surgery is not recommended due to poor outcomes or increased morbidity from combined treatments (e.g., FIGO stages IB3, IIA2, IIB, IIIA-IIIB), the preferred or definitive treatment is concurrent chemoradiotherapy (CRT) followed by internal radiation (brachytherapy). In these cases, the second blood sample will be collected at the end of adjuvant treatment (+14 days).

Inclusion criteria

• Females, over 18 years, with histologically confirmed squamous cell carcinoma or adenocarcinoma of the cervix (stage 1a to 4b)
• Undergo intensive primary surgical treatment, radiotherapy (RT) or chemoradiotherapy (CRT).
• Signed written informed consent

Exclusion criteria

• Unable to provide informed consent
• Serious concomitant systemic disorders incompatible with the study (at the discretion of the investigator)
• Prior malignancies <5 years before inclusion, except for successfully treated keratinocyte skin cancers, or ductal carcinoma in situ.