Predicting response to radical (chemo)radiotherapy with circulating HPV DNA in locally advanced head and neck squamous carcinoma

Jen Y Lee, Isaac Garcia-Murillas, Rosalind J Cutts, David Gonzalez De Castro, Lorna Grove, Tara Hurley, Fuqiang Wang, Christopher Nutting, Katie Newbold, Kevin Harrington, Nicholas Turner, Shreerang Bhide, Jen Y Lee, Isaac Garcia-Murillas, Rosalind J Cutts, David Gonzalez De Castro, Lorna Grove, Tara Hurley, Fuqiang Wang, Christopher Nutting, Katie Newbold, Kevin Harrington, Nicholas Turner, Shreerang Bhide

Abstract

Background: Following chemo-radiotherapy (CCRT) for human papilloma virus positive (HPV+) locally advanced head and neck cancer, patients frequently undergo unnecessary neck dissection (ND) and/or repeated biopsies for abnormal PET-CT, which causes significant morbidity. We assessed the role of circulating HPV DNA in identifying 'true' residual disease.

Methods: We prospectively recruited test (n=55) and validation (n=33) cohorts. HPV status was confirmed by E7 RT-PCR. We developed a novel amplicon-based next generation sequencing assay (HPV16-detect) to detect circulating HPV DNA. Circulating HPV DNA levels post-CCRT were correlated to disease response (PET-CT).

Results: In pre-CCRT plasma, HPV-detect demonstrated 100% sensitivity and 93% specificity, and 90% sensitivity and 100% specificity for the test (27 HPV+) and validation (20 HPV+) cohorts, respectively. Thirty-six out of 37 patients (test and validation cohort) with complete samples-set had negative HPV-detect at end of treatment. Six patients underwent ND (3) and repeat primary site biopsies (3) for positive PET-CT but had no viable tumour. One patient had positive HPV-detect and positive PET-CT and liver biopsy, indicating 100% agreement for HPV-detect and residual cancer.

Conclusions: We demonstrate that HPV16-detect is a highly sensitive and specific test for identification of HPV DNA in plasma at diagnosis. HPV DNA post-treatment correlates with clinical response.

Conflict of interest statement

The authors declare no conflict of interest.

Figures

Figure 1
Figure 1
Comparison of HPV16 status by E7 mRNA, p16 staining and HPV16-detect (NGS) assay in tissue and plasma. (A) There was 100% agreement between the HPV16 status assigned by the ‘gold standard’ E7 mRNA expression and HPV16-detect. (B) Test cohort: HPV16-detect assay demonstrated 100% sensitivity and 92.9% specificity in detecting HPV DNA in plasma compared to tissue. (C) Validation cohort: HPV16-detect assay demonstrated 90% sensitivity and 100% specificity in assigning HPV status in plasma compared to p16 staining in tissue. NPV=negative predictive value; PPV=positive predictive value.
Figure 2
Figure 2
HPV DNA tracking in treatment responsive HPV+ patients using the HPV16-detect assay. HPV DNA levels decreased below threshold at 12-weeks post-treatment in patients receiving C-CRT (n=19, A) and in patients who received induction chemotherapy (IC) and CRT (n=7, B). Red plots in (A) highlight the three patients in (C–E). Residual disease was evident in three patients by 18F-FDG PET-CT (C–E), with negative biopsies and below threshold levels of HPV DNA in the plasma taken at the time of biopsies (PA). PA=post-assessment; post-IC=post-induction chemotherapy. A full colour version of this figure is available at the British Journal of Cancer journal online.
Figure 3
Figure 3
HPV16-detect assay tracked presence of residual disease with 18F-FDG PET-CT at corresponding time points. HPV DNA levels decreased in response to treatment at 6 weeks, but increased significantly at 12 weeks, which corresponded to presence of liver metastases on 18F-FDG PET-CT. HPV DNA levels decreased post-liver resection, however never dropped below the threshold. 18F-FDG PET-CT 8 months following liver surgery revealed disease relapse in cervical lymph nodes. CRT=chemo-radiotherapy; PA=post-assessment; post-IC=post-induction chemotherapy. A full colour version of this figure is available at the British Journal of Cancer journal online.
Figure 4
Figure 4
HPV DNA tracking in the validation cohort with HPV16-detect assay. HPV DNA levels decreased below threshold at 12-weeks post-treatment (n=10, A). Red plots in (A) highlight the three patients in (B–D) with residual lymph node disease on 18F-FDG PET-CT, negative ND specimens and below threshold levels of HPV DNA in the plasma taken at the time ND. PA=post-assessment; post-IC=post-induction chemotherapy; Post-1 week=post-5 fractions of radiotherapy. A full colour version of this figure is available at British Journal of Cancer journal online.

References

    1. Abgral R, Querellou S, Potard G, Le Roux PY, Le Duc-Pennec A, Marianovski R, Pradier O, Bizais Y, Kraeber-Bodere F, Salaun PY (2009) Does 18F-FDG PET/CT improve the detection of posttreatment recurrence of head and neck squamous cell carcinoma in patients negative for disease on clinical follow-up? J Nucl Med 50(1): 24–29.
    1. Ahn SM, Chan JY, Zhang Z, Wang H, Khan Z, Bishop JA, Westra W, Koch WM, Califano JA (2014) Saliva and plasma quantitative polymerase chain reaction-based detection and surveillance of human papillomavirus-related head and neck cancer. JAMA Otolaryngol Head Neck Surg 140(9): 846–854.
    1. Bhide S, Clark C, Harrington K, Nutting CM (2007) Intensity modulated radiotherapy improves target coverage and parotid gland sparing when delivering total mucosal irradiation in patients with squamous cell carcinoma of head and neck of unknown primary site. Med Dosim 32(3): 188–195.
    1. Blanchard P, Bourhis J, Lacas B, Posner MR, Vermorken JB, Hernandez JJ, Bourredjem A, Calais G, Paccagnella A, Hitt R, Pignon JP Meta-Analysis of Chemotherapy in H, Neck Cancer IPCG (2013) Taxane-cisplatin-fluorouracil as induction chemotherapy in locally advanced head and neck cancers: an individual patient data meta-analysis of the meta-analysis of chemotherapy in head and neck cancer group. J Clin Oncol 31(23): 2854–2860.
    1. Burk RD, Harari A, Chen Z (2013) Human papillomavirus genome variants. Virology 445(1-2): 232–243.
    1. Cao H, Banh A, Kwok S, Shi X, Wu S, Krakow T, Khong B, Bavan B, Bala R, Pinsky BA, Colevas D, Pourmand N, Koong AC, Kong CS, Le QT (2012) Quantitation of human papillomavirus DNA in plasma of oropharyngeal carcinoma patients. Int J Radiat Oncol Biol Phys 82(3): e351–e358.
    1. Cullen M, Boland JF, Schiffman M, Zhang X, Wentzensen N, Yang Q, Chen Z, Yu K, Mitchell J, Roberson D, Bass S, Burdette L, Machado M, Ravichandran S, Luke B, Machiela MJ, Andersen M, Osentoski M, Laptewicz M, Wacholder S, Feldman A, Raine-Bennett T, Lorey T, Castle PE, Yeager M, Burk RD, Mirabello L (2015) Deep sequencing of HPV16 genomes: A new high-throughput tool for exploring the carcinogenicity and natural history of HPV16 infection. Papillomavirus Res 1: 3–11.
    1. Dahlstrom KR, Li G, Hussey CS, Vo JT, Wei Q, Zhao C, Sturgis EM (2015) Circulating human papillomavirus DNA as a marker for disease extent and recurrence among patients with oropharyngeal cancer. Cancer 121(19): 3455–3464.
    1. Dillon MT, Harrington KJ (2015) Human papillomavirus-negative pharyngeal cancer. J Clin Oncol 33(29): 3251–3261.
    1. Garcia-Murillas I, Schiavon G, Weigelt B, Ng C, Hrebien S, Cutts RJ, Cheang M, Osin P, Nerurkar A, Kozarewa I, Garrido JA, Dowsett M, Reis-Filho JS, Smith IE, Turner NC (2015) Mutation tracking in circulating tumor DNA predicts relapse in early breast cancer. Sci Transl Med 7(302): 302ra133.
    1. Hamoir M, Ferlito A, Schmitz S, Hanin FX, Thariat J, Weynand B, Machiels JP, Gregoire V, Robbins KT, Silver CE, Strojan P, Rinaldo A, Corry J, Takes RP (2012) The role of neck dissection in the setting of chemoradiation therapy for head and neck squamous cell carcinoma with advanced neck disease. Oral Oncol 48(3): 203–210.
    1. Hopman AH, Smedts F, Dignef W, Ummelen M, Sonke G, Mravunac M, Vooijs GP, Speel EJ, Ramaekers FC (2004) Transition of high-grade cervical intraepithelial neoplasia to micro-invasive carcinoma is characterized by integration of HPV 16/18 and numerical chromosome abnormalities. J Pathol 202(1): 23–33.
    1. Huang SH, O’Sullivan B, Xu W, Zhao H, Chen DD, Ringash J, Hope A, Razak A, Gilbert R, Irish J, Kim J, Dawson LA, Bayley A, Cho BC, Goldstein D, Gullane P, Yu E, Perez-Ordonez B, Weinreb I, Waldron J (2013) Temporal nodal regression and regional control after primary radiation therapy for N2-N3 head-and-neck cancer stratified by HPV status. Int J Radiat Oncol Biol Phys 87(5): 1078–1085.
    1. Jeannot E, Becette V, Campitelli M, Calméjane M-A, Lappartient E, Ruff E, Saada S, Holmes A, Bellet D, Sastre-Garau X (2016) Circulating human papillomavirus DNA detected using droplet digital PCR in the serum of patients diagnosed with early stage human papillomavirus-associated invasive carcinoma. J Pathol Clin Res 2(4): 201–209.
    1. Lamarcq L, Deeds J, Ginzinger D, Perry J, Padmanabha S, Smith-McCune K (2002) Measurements of human papillomavirus transcripts by real time quantitative reverse transcription-polymerase chain reaction in samples collected for cervical cancer screening. J Mol Diagn 4(2): 97–102.
    1. Li H, Handsaker B, Wysoker A, Fennell T, Ruan J, Homer N, Marth G, Abecasis G, Durbin R Genome Project Data Processing S (2009) The Sequence Alignment/Map format and SAMtools. Bioinformatics 25(16): 2078–2079.
    1. Masterson L, Moualed D, Liu ZW, Howard JE, Dwivedi RC, Tysome JR, Benson R, Sterling JC, Sudhoff H, Jani P, Goon PK (2014) De-escalation treatment protocols for human papillomavirus-associated oropharyngeal squamous cell carcinoma: a systematic review and meta-analysis of current clinical trials. Eur J Cancer 50(15): 2636–2648.
    1. Mazurek AM, Rutkowski T, Fiszer-Kierzkowska A, Malusecka E, Skladowski K (2016) Assessment of the total cfDNA and HPV16/18 detection in plasma samples of head and neck squamous cell carcinoma patients. Oral Oncol 54: 36–41.
    1. Mehanna H, Beech T, Nicholson T, El-Hariry I, McConkey C, Paleri V, Roberts S (2013) Prevalence of human papillomavirus in oropharyngeal and nonoropharyngeal head and neck cancer—systematic review and meta-analysis of trends by time and region. Head Neck 35(5): 747–755.
    1. Mehanna H, Wong WL, McConkey CC, Rahman JK, Robinson M, Hartley AG, Nutting C, Powell N, Al-Booz H, Robinson M, Junor E, Rizwanullah M, von Zeidler SV, Wieshmann H, Hulme C, Smith AF, Hall P, Dunn J Group P-NTM (2016) PET-CT surveillance versus neck dissection in advanced head and neck cancer. N Engl J Med 374(15): 1444–1454.
    1. Quinlan AR, Hall IM (2010) BEDTools: a flexible suite of utilities for comparing genomic features. Bioinformatics 26(6): 841–842.
    1. Rettig EM, Wentz A, Posner MR, Gross ND, Haddad RI, Gillison ML, Fakhry C, Quon H, Sikora AG, Stott WJ, Lorch JH, Gourin CG, Guo Y, Xiao W, Miles BA, Richmon JD, Andersen PE, Misiukiewicz KJ, Chung CH, Gerber JE, Rajan SD, D’Souza G (2015) Prognostic implication of persistent human Papillomavirus type 16 DNA detection in oral rinses for human papillomavirus-related oropharyngeal carcinoma. JAMA Oncol 1(7): 907–915.
    1. Schache AG, Liloglou T, Risk JM, Filia A, Jones TM, Sheard J, Woolgar JA, Helliwell TR, Triantafyllou A, Robinson M, Sloan P, Harvey-Woodworth C, Sisson D, Shaw RJ (2011) Evaluation of human papilloma virus diagnostic testing in oropharyngeal squamous cell carcinoma: sensitivity, specificity, and prognostic discrimination. Clin Cancer Res 17(19): 6262–6271.
    1. Schache AG, Powell NG, Cuschieri KS, Robinson M, Leary S, Mehanna H, Rapozo D, Long A, Cubie H, Junor E, Monaghan H, Harrington KJ, Nutting CM, Schick U, Lau AS, Upile N, Sheard J, Brougham K, West CM, Oguejiofor K, Thomas S, Ness AR, Pring M, Thomas GJ, King EV, McCance DJ, James JA, Moran M, Sloan P, Shaw RJ, Evans M, Jones TM (2016) HPV-related oropharynx cancer in the United Kingdom: an evolution in the understanding of disease etiology. Cancer Res 76(22): 6598–6606.
    1. Shah S, Har-El G, Rosenfeld RM (2001) Short-term and long-term quality of life after neck dissection. Head Neck 23(11): 954–961.
    1. Wang Y, Springer S, Mulvey CL, Silliman N, Schaefer J, Sausen M, James N, Rettig EM, Guo T, Pickering CR, Bishop JA, Chung CH, Califano JA, Eisele DW, Fakhry C, Gourin CG, Ha PK, Kang H, Kiess A, Koch WM, Myers JN, Quon H, Richmon JD, Sidransky D, Tufano RP, Westra WH, Bettegowda C, Diaz LA Jr., Papadopoulos N, Kinzler KW, Vogelstein B, Agrawal N (2015) Detection of somatic mutations and HPV in the saliva and plasma of patients with head and neck squamous cell carcinomas. Sci Transl Med 7(293): 293ra104.
    1. Wilkins AC, Rosenfelder N, Schick U, Gupta S, Thway K, Nutting CM, Harrington KJ, Newbold K, Bhide SA (2013) Equivalence of cisplatin and carboplatin-based chemoradiation for locally advanced squamous cell carcinoma of the head and neck: a matched-pair analysis. Oral Oncol 49(6): 615–619.

Source: PubMed

Sponsorit ja yhteistyökumppanit

Sairaudet

Huumeiden interventiot

3
Tilaa