Prognostic Implications of HPV Cell-Free DNA Serial Testing During Follow-Up of p16 Positive Oropharyngeal Squamous Cell Carcinoma After Curative-Intent Treatment

V. Salati; M. Adamowicz; L McKean; D. Noble; D Srinivasan; J MacKenzie; S Linton; C. Callaghan; C. Robert; K. Cuschieri; B. Conn; A. Hay; T.J. Aitman; I.J. Nixon

doi:10.1016/j.clon.2025.103807

Prognostic Implications of HPV Cell-Free DNA Serial Testing During Follow-Up of p16 Positive Oropharyngeal Squamous Cell Carcinoma After Curative-Intent Treatment

V. Salati, M. Adamowicz, L McKean, D. Noble, D Srinivasan, J MacKenzie, S Linton, C. Callaghan, C. Robert, K. Cuschieri, B. Conn, A. Hay, T.J. Aitman^*, I.J. Nixon

^*Corresponding author for this work

Research output: Contribution to journal › Article › peer-review

Abstract

Introduction: Plasma circulating HPV cell-free DNA has high sensitivity and specificity for the detection of HPV-mediated oropharyngeal squamous cell carcinoma. We investigated the clinical significance of serial testing after curative-intent treatments.
Materials and Methods: Patients with concordant p16 positive tumour or neck node biopsy and positive high-risk HPV plasma cell-free DNA were prospectively recruited. HPV cell-free DNA were obtained using digital droplet polymerase chain reaction (ddPCR) and were collected at diagnosis and at every clinical follow-up. Three months after completion of curative-intent treatments, patients were stratified according to treatment response on computed tomography. Complete responders (CR) were followed-up clinically, partial responders (PR) underwent further imaging and surgical/medical management if appropriate, patients with
progressive disease (PD) received palliative treatments.
Results: A hundred and fourteen patients were included and 717 HPV cfDNA ddPCR samples were analysed during a median follow-up of 103 weeks (IQR, 40.2e147.8). Ninety (78.9%) patients were classified as CR, 18 (15.8%) as PR and all except one, who was rapidly diagnosed with PD, had negative HPV ddPCR at 12 weeks follow-up; 6 (5.3%) had PD and all except one had positive HPV ddPCR. Eleven had recurrent disease, 6 in the CR group (6.6%) and 5 among PR (27.7%).
Ninety patients had consistently negative HPV ddPCR at all time points and one developed a recurrence (NPV 99%, 95% C.I., 93.2e99.8%). Eighteen patients developed positive HPV ddPCR and 10 developed recurrent disease (PPV 55%, 95% C.I., 38.6e71.4%). Ten patients had two consecutively positive HPV ddPCR and all had proven disease (PPV 100%, 95% C.I., 69.2e100%). Nine patients had transiently positive HPV ddPCR and none developed disease at that time.
Conclusions: Post-treatment HPV ddPCR reflected treatment response on imaging and serial testing had high PPV and NPV in detecting recurrent disease.

Original language	English
Article number	103807
Journal	Clinical Oncology
Volume	41
Early online date	19 Mar 2025
DOIs	https://doi.org/10.1016/j.clon.2025.103807
Publication status	Published - 1 May 2025

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https://linkinghub.elsevier.com/retrieve/pii/S0936655525000627

Comprehensive longitudinal characterisation of refractory and relapsing oropharyngeal squamous cell carcinoma by tumour and blood-based genome sequence analysis
Aitman, T. (Principal Investigator) & Thomson, J. (Co-investigator)
Scottish Executive
1/05/21 → 31/05/24
Project: Research

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Salati, V., Adamowicz, M., McKean, L., Noble, D., Srinivasan, D., MacKenzie, J., Linton, S., Callaghan, C., Robert, C., Cuschieri, K., Conn, B., Hay, A., Aitman, T. J., & Nixon, I. J. (2025). Prognostic Implications of HPV Cell-Free DNA Serial Testing During Follow-Up of p16 Positive Oropharyngeal Squamous Cell Carcinoma After Curative-Intent Treatment. Clinical Oncology, 41, Article 103807. https://doi.org/10.1016/j.clon.2025.103807

@article{a0f29f63d6794e249d0a32f93548c78c,

title = "Prognostic Implications of HPV Cell-Free DNA Serial Testing During Follow-Up of p16 Positive Oropharyngeal Squamous Cell Carcinoma After Curative-Intent Treatment",

abstract = "Introduction: Plasma circulating HPV cell-free DNA has high sensitivity and specificity for the detection of HPV-mediated oropharyngeal squamous cell carcinoma. We investigated the clinical significance of serial testing after curative-intent treatments. Materials and Methods: Patients with concordant p16 positive tumour or neck node biopsy and positive high-risk HPV plasma cell-free DNA were prospectively recruited. HPV cell-free DNA were obtained using digital droplet polymerase chain reaction (ddPCR) and were collected at diagnosis and at every clinical follow-up. Three months after completion of curative-intent treatments, patients were stratified according to treatment response on computed tomography. Complete responders (CR) were followed-up clinically, partial responders (PR) underwent further imaging and surgical/medical management if appropriate, patients with progressive disease (PD) received palliative treatments. Results: A hundred and fourteen patients were included and 717 HPV cfDNA ddPCR samples were analysed during a median follow-up of 103 weeks (IQR, 40.2e147.8). Ninety (78.9\%) patients were classified as CR, 18 (15.8\%) as PR and all except one, who was rapidly diagnosed with PD, had negative HPV ddPCR at 12 weeks follow-up; 6 (5.3\%) had PD and all except one had positive HPV ddPCR. Eleven had recurrent disease, 6 in the CR group (6.6\%) and 5 among PR (27.7\%). Ninety patients had consistently negative HPV ddPCR at all time points and one developed a recurrence (NPV 99\%, 95\% C.I., 93.2e99.8\%). Eighteen patients developed positive HPV ddPCR and 10 developed recurrent disease (PPV 55\%, 95\% C.I., 38.6e71.4\%). Ten patients had two consecutively positive HPV ddPCR and all had proven disease (PPV 100\%, 95\% C.I., 69.2e100\%). Nine patients had transiently positive HPV ddPCR and none developed disease at that time. Conclusions: Post-treatment HPV ddPCR reflected treatment response on imaging and serial testing had high PPV and NPV in detecting recurrent disease.",

author = "V. Salati and M. Adamowicz and L McKean and D. Noble and D Srinivasan and J MacKenzie and S Linton and C. Callaghan and C. Robert and K. Cuschieri and B. Conn and A. Hay and T.J. Aitman and I.J. Nixon",

year = "2025",

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doi = "10.1016/j.clon.2025.103807",

language = "English",

volume = "41",

journal = "Clinical Oncology",

issn = "0936-6555",

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Salati, V, Adamowicz, M, McKean, L, Noble, D, Srinivasan, D, MacKenzie, J, Linton, S, Callaghan, C, Robert, C , Cuschieri, K, Conn, B, Hay, A, Aitman, TJ & Nixon, IJ 2025, 'Prognostic Implications of HPV Cell-Free DNA Serial Testing During Follow-Up of p16 Positive Oropharyngeal Squamous Cell Carcinoma After Curative-Intent Treatment', Clinical Oncology, vol. 41, 103807. https://doi.org/10.1016/j.clon.2025.103807

TY - JOUR

T1 - Prognostic Implications of HPV Cell-Free DNA Serial Testing During Follow-Up of p16 Positive Oropharyngeal Squamous Cell Carcinoma After Curative-Intent Treatment

AU - Salati, V.

AU - Adamowicz, M.

AU - McKean, L

AU - Noble, D.

AU - Srinivasan, D

AU - MacKenzie, J

AU - Linton, S

AU - Callaghan, C.

AU - Robert, C.

AU - Cuschieri, K.

AU - Conn, B.

AU - Hay, A.

AU - Aitman, T.J.

AU - Nixon, I.J.

PY - 2025/5/1

Y1 - 2025/5/1

N2 - Introduction: Plasma circulating HPV cell-free DNA has high sensitivity and specificity for the detection of HPV-mediated oropharyngeal squamous cell carcinoma. We investigated the clinical significance of serial testing after curative-intent treatments. Materials and Methods: Patients with concordant p16 positive tumour or neck node biopsy and positive high-risk HPV plasma cell-free DNA were prospectively recruited. HPV cell-free DNA were obtained using digital droplet polymerase chain reaction (ddPCR) and were collected at diagnosis and at every clinical follow-up. Three months after completion of curative-intent treatments, patients were stratified according to treatment response on computed tomography. Complete responders (CR) were followed-up clinically, partial responders (PR) underwent further imaging and surgical/medical management if appropriate, patients with progressive disease (PD) received palliative treatments. Results: A hundred and fourteen patients were included and 717 HPV cfDNA ddPCR samples were analysed during a median follow-up of 103 weeks (IQR, 40.2e147.8). Ninety (78.9%) patients were classified as CR, 18 (15.8%) as PR and all except one, who was rapidly diagnosed with PD, had negative HPV ddPCR at 12 weeks follow-up; 6 (5.3%) had PD and all except one had positive HPV ddPCR. Eleven had recurrent disease, 6 in the CR group (6.6%) and 5 among PR (27.7%). Ninety patients had consistently negative HPV ddPCR at all time points and one developed a recurrence (NPV 99%, 95% C.I., 93.2e99.8%). Eighteen patients developed positive HPV ddPCR and 10 developed recurrent disease (PPV 55%, 95% C.I., 38.6e71.4%). Ten patients had two consecutively positive HPV ddPCR and all had proven disease (PPV 100%, 95% C.I., 69.2e100%). Nine patients had transiently positive HPV ddPCR and none developed disease at that time. Conclusions: Post-treatment HPV ddPCR reflected treatment response on imaging and serial testing had high PPV and NPV in detecting recurrent disease.

AB - Introduction: Plasma circulating HPV cell-free DNA has high sensitivity and specificity for the detection of HPV-mediated oropharyngeal squamous cell carcinoma. We investigated the clinical significance of serial testing after curative-intent treatments. Materials and Methods: Patients with concordant p16 positive tumour or neck node biopsy and positive high-risk HPV plasma cell-free DNA were prospectively recruited. HPV cell-free DNA were obtained using digital droplet polymerase chain reaction (ddPCR) and were collected at diagnosis and at every clinical follow-up. Three months after completion of curative-intent treatments, patients were stratified according to treatment response on computed tomography. Complete responders (CR) were followed-up clinically, partial responders (PR) underwent further imaging and surgical/medical management if appropriate, patients with progressive disease (PD) received palliative treatments. Results: A hundred and fourteen patients were included and 717 HPV cfDNA ddPCR samples were analysed during a median follow-up of 103 weeks (IQR, 40.2e147.8). Ninety (78.9%) patients were classified as CR, 18 (15.8%) as PR and all except one, who was rapidly diagnosed with PD, had negative HPV ddPCR at 12 weeks follow-up; 6 (5.3%) had PD and all except one had positive HPV ddPCR. Eleven had recurrent disease, 6 in the CR group (6.6%) and 5 among PR (27.7%). Ninety patients had consistently negative HPV ddPCR at all time points and one developed a recurrence (NPV 99%, 95% C.I., 93.2e99.8%). Eighteen patients developed positive HPV ddPCR and 10 developed recurrent disease (PPV 55%, 95% C.I., 38.6e71.4%). Ten patients had two consecutively positive HPV ddPCR and all had proven disease (PPV 100%, 95% C.I., 69.2e100%). Nine patients had transiently positive HPV ddPCR and none developed disease at that time. Conclusions: Post-treatment HPV ddPCR reflected treatment response on imaging and serial testing had high PPV and NPV in detecting recurrent disease.

U2 - 10.1016/j.clon.2025.103807

DO - 10.1016/j.clon.2025.103807

M3 - Article

SN - 0936-6555

VL - 41

JO - Clinical Oncology

JF - Clinical Oncology

M1 - 103807

ER -

Prognostic Implications of HPV Cell-Free DNA Serial Testing During Follow-Up of p16 Positive Oropharyngeal Squamous Cell Carcinoma After Curative-Intent Treatment

Abstract

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Comprehensive longitudinal characterisation of refractory and relapsing oropharyngeal squamous cell carcinoma by tumour and blood-based genome sequence analysis

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