Key differences between HPV-positive and HPV-negative head and neck squamous cell carcinomas (HNSCC)

Head and neck squamous cell carcinomas (HNSCC) develop from the mucosal epithelium in the oral cavity, pharynx, and larynx and are the most common malignancies that arise in the head and neck.1 Increasingly, tumors in the oropharynx are linked to prior infection with oncogenic strains of human papillomavirus (HPV), primarily HPV-16 and, to a lesser extent HPV-18 and others.2,3 HNSCCs of the oral cavity and larynx are primarily associated with tobacco-derived carcinogens, excessive alcohol consumption, or both, and are collectively referred to as HPV-negative HNSCC.4 The median age of diagnosis for non-virally associated HNSCC is 66 years, whereas the median age of diagnosis for HPV-associated oropharyngeal cancer is about 53 years.5

HPV-positive and HPV-negative HNSCC are two distinct diseases with different sites of origin, etiological agents, risk factors, and contributions to the development of oropharyngeal squamous cell carcinoma (OPSCC). The demographics, cause, and prevalence of HPV-positive and NPV-negative cancers are summarized in Table 1.

Table 1: Demographics, Cause, and Prevalence of HNSCC.

Whereas the incidence of smoking-related HNSCC continues to decline worldwide, that of HPV-positive HNSCC is on the rise.6 During 2007-2016, HPV-associated cancers increased by 2.1% per year on average, whereas cancers not associated with HPV decreased by 0.4% per year on average.6

HPV-positive and HPV-negative HNSCCs present with different molecular characteristics, immune landscapes, and clinical prognosis (Table 2) and lead to two fundamentally different diseases with distinct pathogenesis in terms of gene expression, tumor microenvironment (TME), and mutational burden.

Table 2. Pathology and Disease Signature in HNSCC.

Genomic and epigenetic analyses reveal extremely high heterogeneity in HNSCC in terms of characteristic mutations, molecular signature, cellular phenotype, composition of TME, and immune landscape (Table 3).

Table 3. Characteristic mutations in HPV-positive and HPV-negative HNSCC.

Various promising vaccine targets have been identified and treatment options employed for the treatment of HPV-positive and HPV-negative cancers with varying degrees of success (Table 4).

Table 4. Treatment options and targets.
Treatment personalization and de-escalation

According to Dr. Nishant Agrawal, Chief of Otolaryngology-Head & Neck Surgery UChicago Medicine, “We have seen a significant increase in the incidence of HPV-associated oropharyngeal cancer in relatively younger patients, with the median age of diagnosis in the 50s, even patients in their 30s.”32 He added that “Even at 2 years after radiation therapy, 15% of patients had grade 2 swallowing dysfunction and 8% had progressive dysphagia, so their swallowing is going to continue to get worse. Patients may also have chronic xerostomia. The dry mouth improves but it never gets back to 100%.”33 In contrast to patients with HPV-negative HNSCC, who have a five-year survival rate of about 25%-40%, patients with HPV-positive HNSCC fare much better with a disease-free survival rate of 85%-90% over five years. According to Dr. Agrawal, the better prognosis for HPV-positive patients suggests a need to de-escalate treatment while preserving survival.

References

  1. Stein, A.P. et al. (2015) Prevalence of human papillomavirus in oropharyngeal cancer: a systematic review. Cancer J. (21);138-46.
  2. Isayeva, T. et al. (2012) Human papillomavirus in non-oropharyngeal head and neck cancers: a systematic literature review. Head Neck Pathol. (6);S104-20.
  3. Michaud, D.S. et al. (2014) High-risk HPV types and head and neck cancer. Int J Cancer. (135);1653-61.
  4. https://pubmed.ncbi.nlm.nih.gov/33243986/  
  5. Windon, M.J. et al. (2018) Increasing prevalence of human papillomavirus-positive oropharyngeal cancers among older adults. Cancer (124);2993-99.
  6. Ellington, T.D. et al. (2020) Trends in incidence of cancers of the oral cavity and pharynx – United States 2007-2016. Morb Morta Wkly Rep. (69);433-38.
  7. https://www.cancer.org/cancer/oral-cavity-and-oropharyngeal-cancer/causes-risks-prevention/risk-factors.html  
  8. https://pubmed.ncbi.nlm.nih.gov/33243986/  
  9. Gillison, M.L. et al. (2015) Epidemiology of human papillomavirus positive head and neck squamous cell carcinoma. J Clin Oncol. (33);3235-42.
  10. Mahal, B.A. et al. (2019) Incidence and Demographic Burden of HPV-Associated Oropharyngeal Head and Neck Cancers in the United States. Cancer Epidemiol Biomark Prev. 28(10);1660-67.
  11. Kazuhiro, K. et al. (2018) A Review of HPV-Related Head and Neck Cancer. J Clin Med Sep. 7(9);241.
  12. https://bmccancer.biomedcentral.com/articles/10.1186/s12885-022-09407-5 
  13. https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5486734
  14. Canning, M. et al. (2019) Heterogeneity of the Head and Neck Squamous Cell Carcinoma Immune Landscape and Its Impact on Immunotherapy. Front Cell Dev Bio.l vol7. https://doi.org/10.3389/fcell.2019.00052
  15. Fakhry, C. et al. (2017) The prognostic role of sex, race, and human papillomavirus in oropharyngeal and nonoropharyngeal head and neck squamous cell cancer. Cancer (123);1566-75. Doi: 10.1002/cncr.30353.
  16. Pai, S.I. et al. (2009) Molecular pathology of head and neck cancer: implications for diagnosis, prognosis, and treatment. Annu Rev Pathol. (4);49-70.
  17. Keck, M.K. et al. (2015) Integrative analysis of head and neck cancer identifies two biologically distinct HPV and three non-HPV subtypes. Clin Cancer Res. (21);870-81. Doi: 10.1158/1078-0432.CCR-14-2481.
  18. Hanna, G.J. et al. (2018) Frameshift events predict anti-PD-1/L1 response in head and neck cancer. JCI Insight 3:98811. Doi: 10.1172/jci.insight.98811.
  19. Elpek, K.G. et al. (2014) The tumor microenvironment shapes lineage, transcriptional, and functional diversity of infiltrating myeloid cells. Cancer Immunol Res. (2);655-67. Doi: 10.1158/2326-6066.CIR-13-0209.
  20. Mandal, R. et al. (2016) The head and neck cancer immune landscape and its immunotherapeutic implications. JCI Insight 1: e89829. Doi: 10.1172/jci.insight.89829.
  21. Badoual, C. et al. (2013) PD-1-expressing tumor-infiltrating T cells are a favorable prognostic biomarker in HPV-associated head and neck cancer. Cancer Res. (73);128-38. Doi: 10.1158/0008-5472.CAN-12-2606.
  22. Hanna, G.J. et al. (2017) Defining an inflamed tumor immunophenotype in recurrent, metastatic squamous cell carcinoma of the head and neck. Oral Oncol. (67);61-69.
  23. Taberna, M. et al. (2017) Human papillomavirus-related oropharyngeal cancer. Ann Oncol. (28);2386-98.
  24. Coca-Pelaz, A. et al. (2020) The risk of second primary tumors in head and neck cancer: a systematic review. Head Neck. (42);456-66.
  25. Tomaic, V. (2016) Functional roles of E6 and E7 oncoproteins in HPV-induced malignancies at diverse anatomical sites. Cancers (8);95.
  26. Johnson, D.E. et al. (2020) Head and neck squamous cell carcinoma. Nat Rev Dis Primers. (6);92.
  27. The Cancer Genome Atlas Network [TCGA]. (2015) Comprehensive genomic characterization of head and neck squamous cell carcinomas. Nature (517) 576-82.
  28. Beck, T.N. et al. (2016) EGFR and RB1 as Dual Biomarkers in HPV-Negative Head and Neck Cancer. Mol Cancer Ther. 15(10);2486-97.
  29. Schreiber, R.D. et al. (2011) Cancer immunoediting: integrating immunity’s roles in cancer suppression and promotion. Science 331;1565-70.
  30. Skeate, J.G., et al. (2016) Current therapeutic vaccination and immunotherapy strategies for HPV-related diseases. Hum Vaccin Immunother. (12);1418-29.
  31. Seiwert, T.Y. et al. (2016) Safety and clinical activity of pembrolizumab for treatment of recurrent or metastatic squamous cell carcinoma of the head and neck (KEYNOTE-012): an open-label, multicentre, phase 1b trial. Lancet Oncol. (17);956-65.
  32. Dr. Nishant Agrawal, Chief of Otolaryngology-Head & Neck Surgery UChicago Medicine. Personal interview on 3/25/22.
  33. https:\\ascopost.com\\issues\\april-25-2017\\deintensifiying-treatment-of-hpv-positive-oropharyngeal-cancer-could-reduce-toxicity-while-maintaining-function-and-survival

Sysmex Inostics & QIAGEN Highlight Cancer Companion Diagnostics Alliance at ASCO 2022

Two poster presentations utilizing Sysmex Inostics technology to be featured

Baltimore, MD, June 2, 2022 – Sysmex Inostics, Inc. and QIAGEN N.V. will co-exhibit at the 2022 ASCO Annual Meeting, the world’s largest clinical cancer research meeting, held June 3, 2022, through June 7, 2022, in Chicago.

QIAGEN, the leading global provider of sample to insight solutions for molecular testing, and Sysmex Inostics combined forces in July 2021 to accelerate global companion diagnostic access. QIAGEN provides unparalleled global custom cancer companion diagnostics (CDx) development and commercialization capabilities utilizing Sysmex Inostics ultra-sensitive NGS liquid biopsy technology. The goal of the alliance is to promote early clinical implementation of Sysmex Inostics’ technology to expedite clinical trial timelines for pharmaceutical companies that develop molecularly targeted drugs for cancer.

“Sysmex Inostics is focused on developing tools to support the fight against cancer and other devastating diseases. By partnering with industry leaders like QIAGEN we can help accelerate the fight to help patients with these diseases,” said Shinichi Sato, CEO of Sysmex Inostics, Inc. “We look forward to highlighting our partnership with QIAGEN at ASCO 2022.”

“Through our partnership with Sysmex, we can develop ultrasensitive blood-based NGS panels to the specific requirements of our pharma partners for their clinical trials,” stated Jonathan Arnold, Vice President, Head of Oncology and Precision Diagnostics of QIAGEN. Arnold added, “If and when a CDx is required, QIAGEN will lead IVD submission, manufacturing, and global commercialization as a kitted product.”

Sysmex and QIAGEN will highlight their alliance at ASCO booth 8149, Saturday, June 4, 2022, through Monday, June 6, 2022, 9:00 AM – 5:00 PM CDT each day.

Poster Presentations Utilizing Sysmex Inostics’ Technology
Yoshinori Kagawa MD, PhD, Department of Gastroenterological Surgery, Osaka General Medical Center, Osaka, Japan, will present the poster “Plasma RAS dynamics and anti-EGFR rechallenge efficacy in patients with RAS/BRAF wild-type metastatic colorectal cancer: REMARRY and PURSUIT trials.” Circulating tumor DNA (ctDNA) was analyzed and monitored using the Sysmex Inostics OncoBEAMTM RAS CRC Kit. Dr. Kagawa will present poster 3518 during the Gastrointestinal Cancer—Colorectal and Anal session on Saturday, June 4, 2022, 3:00 PM-4:30 PM CDT.

Linda (Yilin) Cao, MD, Department of Radiation Oncology, Sidney Kimmel Comprehensive Cancer Center at Johns Hopkins, will present the poster “Dynamic cell free HPV DNA is an early measure of treatment responsiveness in patients receiving induction chemotherapy for HPV-related head and neck cancer.” Findings add to the breadth of ongoing clinical studies using HPV-SEQ to investigate de-escalation of HPV-positive patients from unnecessary and potentially harmful head and neck squamous cell carcinomas (HNSCC) treatments. HPV-SEQ is an ultrasensitive test for the detection of HPV16/18 DNA from blood and is CLIA validated to detect as few as five HPV DNA molecules from two tubes of blood.1 See here for additional study information. Dr. Cao will present poster 6062 during the Head and Neck Cancer session on Monday, June 6, 2022, 1:15 PM- 4:15 PM CDT.

Posters will be available after each presentation here:
Plasma RAS dynamics and anti-EGFR rechallenge efficacy
Dynamic cell free HPV DNA is an early measure of treatment responsiveness

About QIAGEN
QIAGEN N.V., a Netherlands-based holding company, is the leading global provider of Sample to Insight solutions that enable customers to gain valuable molecular insights from samples containing the building blocks of life. Our sample technologies isolate and process DNA, RNA and proteins from blood, tissue and other materials. Assay technologies make these biomolecules visible and ready for analysis. Bioinformatics software and knowledge bases interpret data to report relevant, actionable insights. Automation solutions tie these together in seamless and cost-effective workflows. QIAGEN provides solutions to more than 500,000 customers around the world in Molecular Diagnostics (human healthcare), Applied Testing (primarily forensics), Pharma (pharma and biotech companies) and Academia (life sciences research). As of March 31, 2022, QIAGEN employed more than 6,000 people in over 35 locations worldwide. Further information can be found at http://www.qiagen.com.

About Sysmex Inostics
Sysmex Inostics, Inc., a US-based Sysmex Corporation subsidiary, empowers discoveries in oncology by providing investigators ultra-sensitive quantitative liquid biopsy solutions.

Developed by experts at Johns Hopkins with the philosophy of “no molecule left behind,” these technologies are optimized to ensure the detection of low-frequency mutant molecules (0.03%-0.05% MAF) with a high degree of specificity. Focused and flexible genomic coverage allows for superior sensitivity and reduced costs.

As forerunners of blood-based circulating tumor DNA (ctDNA) mutation detection, Sysmex Inostics has provided custom assays and CLIA-certified lab services to leading BioPharma companies over the last ten years to help monitor progression, identify targetable resistance alterations, and detect MRD throughout the clinical trial process.

Sysmex Inostics offers a portfolio of highly sensitive NGS panels through its CLIA-certified laboratory in Baltimore, Maryland.

For more information, refer to www.sysmex-inostics.com or email info@sysmex-inostics.com.

Contact:
Tracy Vandenbroek
Senior Director, Marketing & Strategy
Sysmex Inostics
+1.512.791.2899
vandenbroek.tracy@sysmex-inostics.com

Reference

  1. Internal validation data on file, Sysmex Inostics 2022

Sysmex Inostics Expands Blood Test Offerings for Detection of Acute Myeloid Leukemia

Joins Foundation for the National Institutes of Health Biomarkers Consortium 


Baltimore, MD, May 24, 2022 – Sysmex Inostics, a global leader in the liquid biopsy revolution, has expanded its offering of CLIA-validated tests to include AML-SEQ™, a focused panel to detect mutations of the three most prevalent genes found in Acute Myeloid Leukemia (AML) – IDH1/2 & NPM1. 

 AML-SEQ rounds out the company’s AML Laboratory Developed Test (LDT) offerings with a less expensive test compared to the broader AML-MRD-SEQ panel launched in October 2021. AML-MRD-SEQ is a more extensive panel for the detection of measurable residual disease (MRD) in 68 regions across 20 genes including the clinically established IDH1/2 and NPM1. 

“We envision investigators and eventually physicians using AML-SEQ and AML-MRD-SEQ as a one-two-punch in the fight against AML and AML MRD. We are committed to developing tools to support the fight against this devastating disease. We must support management of patients suffering from AML by providing the accurate and sensitive detection of AML biomarkers,” said Shinichi Sato, CEO of Sysmex Inostics, Inc.

AML-SEQ adds to the portfolio of ultra-sensitive Plasma-Safe-SeqS technology NGS tests available through Sysmex Inostics’ CLIA lab services in Baltimore, Maryland.

Joins Foundation for the National Institutes of Health Biomarkers Consortium

The FNIH consortium combines significant public and private organizations to develop AML MRD guidelines, establish methods for MRD detection, and achieve better measures for clinical trial design and drug development. Project partners include the National Cancer Institute (NCI), the National Heart Lung and Blood Institute (NHLBI), the U.S. Food and Drug Administration (FDA), several award-winning cancer institutions and leading pharmaceutical and diagnostic companies. For more information about the FNIH AML-MRD Consortium click here.

According to the National Cancer Institute (NCI), an estimated 20,050 new AML cases will be diagnosed in the United States in 2022.1 Rising incidence of cancer and increasing preference of noninvasive treatments is driving the quick expansion of the liquid biopsy market. Globally, the liquid biopsy market is to achieve an annual growth rate of more than 18% over the next few years, reaching $5.8 billion by 2026.2

About Sysmex Inostics

Sysmex Inostics, Inc., a US-based Sysmex Corporation subsidiary, empowers discoveries in oncology by providing investigators cost-effective and ultra-sensitive quantitative liquid biopsy solutions.

Developed by experts at Johns Hopkins with the philosophy of “no molecule left behind,” these technologies are optimized to ensure the detection of low-frequency mutant molecules (0.03%-0.05% MAF) with a high degree of specificity. Focused and flexible genomic coverage allows for superior sensitivity and reduced costs.

As forerunners of blood-based circulating tumor DNA (ctDNA) mutation detection, Sysmex Inostics has provided custom assays and CLIA-certified lab services to leading BioPharma companies over the last ten years to help monitor progression, identify targetable resistance alterations, and detect MRD throughout the clinical trial process.

In July 2021, Sysmex Corporation announced a global strategic alliance with QIAGEN to provide custom cancer companion diagnostics (CDx) utilizing Plasma-Safe-SeqS technology. The alliance is intended to promote early clinical implementation of Sysmex Inostics’ technology to expedite clinical trial timelines for pharmaceutical companies that develop molecularly targeted drugs for cancer.

Sysmex Inostics offers a portfolio of highly sensitive NGS panels through its CLIA-certified laboratory in Baltimore, Maryland.

For more information, refer to www.sysmex-inostics.com or email info@sysmex-inostics.com.

Contact:
Tracy Vandenbroek, Senior Director, Marketing & Business Strategy, Sysmex Inostics
+1.512.791.2899
vandenbroek.tracy@sysmex-inostics.com

References

  1.  https://seer.cancer.gov/statfacts/html/amyl.html 
  2.  https://www.marketsandmarkets.com/Market-Reports/liquid-biopsy-market-13966350.html?gclid=CjwKCAiAg6yRBhBNEiwAeVyL0CZs-TXAfcJc_hDJoIHKkdcE7iNCnWH2DIEVxKT0OjDfHTSREKjWBxoCAuYQAvD_BwE 

Sysmex Inostics Introduces CLIA-Validated Highly Sensitive HNSCC-SEQ Testing Services for Head and Neck Cancer at the 2022 Molecular Medicine Tri-Conference

Presenting Poster of Clinical Trial Findings at 2022 Multidisciplinary Head and Neck Cancers Symposium

Baltimore, MD, February 22, 2022 – Sysmex Inostics, a global leader in the liquid biopsy revolution for oncology, will introduce HNSCC-SEQ, a highly sensitive Plasma-Safe-SeqS and Next Generation Sequencing (NGS) assay service for head and neck squamous cell carcinomas (HNSCC), at the annual 2022 Molecular Medicine Tri-Conference being held Monday, February 21st through Wednesday, February 23rd in San Diego, California. The assay has a turn-around-time of 7-10 days and is available to researchers and clinicians.

Sysmex Inostics senior director of medical affairs, Dr. Fred Jones stated, “We see a huge opportunity for researchers and clinicians with our HNSCC-SEQ and HPV-SEQ assays being used in tandem to appropriately identify patients’ tumor mutational drivers.” Jones added, “We know the human papillomavirus (HPV) fuels a growing percentage of head and neck cancers, but there is also an unmet need to track HNSCC tumors via circulating tumor DNA (ctDNA) that are HPV-negative – that’s where HNSCC-SEQ comes in. This HNSCC panel helps researchers and clinicians identify patients quickly and accurately for the appropriate therapy and avoiding over-treatment.”

HNSCC develop from the mucosal tissue in the oral cavity, pharynx, and larynx and are the most common malignancies that arise in the head and neck regions.¹ HNSCC-SEQ was designed for HPV-negative patients and can be used to detect novel therapeutic targets and frequently occurring driver mutations for treatment response monitoring. HNSCC-SEQ delivers high-sensitivity mutation detection in HNSCC with a limit of detection of 0.05% MAF.2

HNSCC-SEQ can identify head and neck cancer mutational drivers from the genes: CDKN2A, EGFR, ERBB2, FGFR3, HRAS, KRAS, NOTCH1, PIK3CA, PTEN, and TP53, many of which are actively being pursued as therapeutic targets.3

Dr. Jones will discuss how Plasma-Safe-SeqS technology, including HNSCC-SEQ, can aid cancer drug development, treatment guidance and monitoring, in addition to post-treatment recurrence monitoring during the 2022 Molecular Medicine Tri- Conference’s C4B- Clinical Biomarkers & Companion Diagnostics presentation track in session room Indigo 206 on Tuesday, February 22nd at the Hilton San Diego Bayfront. More information about the presentation can be viewed here.

2022 Multidisciplinary Head and Neck Cancers Symposium

Additionally, Dr. Ari Rosenberg, Assistant Professor of Medicine at the University of Chicago will present findings from his study using Sysmex Inostics HPV-SEQ test at the 2022 Multidisciplinary Head and Neck Cancers Symposium being held February 24th through 26th in Phoenix, Arizona. The poster titled ‘Dynamic changes of cell-free HPV DNA in locoregional viral-associated oropharyngeal cancer receiving response-adaptive treatment’ will be presented Thursday, February 24th. More information can be viewed here.

HNSCC-SEQ and HPV-SEQ are available as a testing service provided by the Sysmex Inostics CLIA lab in Baltimore. MD.

About Sysmex Inostics

Sysmex Inostics, Inc., a US-based Sysmex Corporation subsidiary, empowers discoveries in oncology by providing investigators with ultra-sensitive, quantitative, and cost-effective CLIA-validated liquid biopsy services.

Since 2008, Sysmex Inostics has provided leading Pharma companies custom liquid biopsy assay services, first OncoBEAM™ and now Plasma-Safe-SeqS technology, to support real-time therapy selection and targeted-mutational monitoring during and after treatment throughout the clinical trial process.

Developed by experts at Johns Hopkins with the philosophy of “no molecule left behind,” Sysmex Inostics Plasma-Safe-SeqS technology has robust detection as low as 0.03% allele frequency (for input of 20,000 genomic equivalents) without sacrificing specificity. The venerable OncoBEAM™ digital PCR cell-free DNA (cfDNA) technology has been employed in hundreds of pivotal studies, publications, and numerous drug discoveries in oncology. Plasma-Safe-SeqS NGS technology, introduced in 2019, is currently being used in various clinical studies.

In July 2021, Sysmex Corporation announced a global strategic alliance with QIAGEN to expedite clinical trial timelines and CDx development by uniting QIAGEN’s commercial and regulatory expertise with the liquid biopsy scientific rigor and knowledge of Sysmex Inostics.

Sysmex Inostics offers Plasma-Safe-SeqS technology services in its CLIA-certified laboratory in Baltimore, Maryland.

For more information, refer to www.sysmex-inostics.com or email info@sysmex-inostics.com.

Contact:
Tracy Vandenbroek Director, Marketing Sysmex Inostics
+1.512.791.2899
vandenbroek.tracy@sysmex-inostics.com

References

  1. Stein, A. P. et al. (2015) Prevalence of human papillomavirus in oropharyngeal cancer: a systematic review. Cancer J. 21, 138-146.
  2. Internal validation data, Sysmex Inostics, Inc.
  3. Jiang X, Ye J, Dong Z, Hu S, Xiao M. Novel genetic alterations and their impact on target therapy response in head and neck squamous cell carcinoma. Cancer Manag Res. 2019;11:1321-1336. Published 2019 Feb 8. doi:10.2147/CMAR.S187780

New ASCO 2021 poster highlights HPV-SEQ test’s ultra-sensitive detection of HPV 16/18 in plasma

Download our new poster, presented at the recent ASCO 2021 Annual Meeting: “Ultra-sensitive detection and quantification of human papillomavirus (HPV) DNA in the plasma of patients with oropharyngeal squamous cell carcinoma (OPSCC) enrolled in the OPTIMA 2 treatment de-escalation trial”.

About HPV-SEQ

As patients with HPV-driven tumors often have a good prognosis, clinical investigators have recently explored new strategies for treatment de-escalation to avoid unnecessary side-effects caused by overtreatment.  Important clinical data for HPV-SEQ was generated while investigating induction chemoimmunotherapy followed by risk/response stratified de-escalated locoregional therapy for patients with HPV+ OPSCC. During the trial, HPV-SEQ was employed to evaluate levels of cfHPV-DNA alongside patients’ radiographic response to therapy to assess the future utility in guiding treatment de-escalation strategies. HPV-SEQ showed robust quantitative detection of HPV 16/18 across a broad dynamic range over five orders of magnitude with low quantitative variability. Importantly, a high correlation was observed between dynamic changes in patients’ cfHPV DNA levels and radiographic responses following induction therapy.