HPV-positive Tumors Research Articles

Therapeutic inhibitory RNA in head and neck cancer via functional targeted lipid nanoparticles

Currently there are no specific therapies addressing the distinctive biology of human papillomavirus (HPV)-induced cancer approved for clinical use. Short interfering RNA (siRNA) has much potential for therapeutic manipulation of HPV E6/E7 oncoproteins. Lipid-based nanoparticles (LNPs) can be utilized for systemic transportation and delivery of siRNA at target site. We recently developed a recombinant protein linker that enables uniform conjugation of targeting antibodies to the LNPs. Herein, we demonstrate the therapeutic efficacy of anti-E6/E7 siRNA delivered via targeted LNPs (tLNPs) in a xenograft HPV-positive tumor model. We show that anti-epidermal growth factor receptor (EGFR) antibodies, anchored to the LNPs as targeting moieties, facilitate cargo delivery but also mediate anti-tumor activity. Treatment with siE6 via tLNPs resulted in 50% greater reduction of tumor volume compared to treatment with siControl encapsulated in isoLNPs (coated with isotype control antibodies). We demonstrate superior suppression of HPV oncogenes and higher induction of apoptosis by the tLNPs both in vitro and in vivo. Altogether, the coupling of inhibitory siE6 with anti-EGFR antibodies, that further elicited anti-tumor effects, successfully restricted tumor progression. This system that combines potent siRNA and therapeutically functional tLNPs can be modulated against various cancer models.

Biologic and behavioral associations of estrogen receptor alpha positivity in head and neck squamous cell carcinoma

ObjectivesTumor HPV status is an established independent prognostic marker for oropharynx cancer (OPC). Recent studies have reported that tumor estrogen receptor alpha (ERα) positivity is also associated with prognosis independent of HPV. Little is known about the biologic and behavioral predictors of ERα positivity in head and neck squamous cell cancer (HNSCC). We therefore explored this in a multicenter prospective cohort study. Materials and MethodsParticipants with HNSCC completed a survey and provided a blood sample. Tumor samples were tested for ERα using immunohistochemistry. ERα positivity was defined as ≥1%, standardized by the American Society of Clinical Oncology/College of American Pathologists in breast cancer. Characteristics were compared with χ2 and Fisher’s exact test. Odds ratios (OR) were calculated using logistic regression. ResultsOf 318 patients with HNSCC, one third had ERα positive tumors (36.2%, n = 115). Odds of ERα expression were significantly increased in those with HPV-positive tumors (OR = 27.5, 95% confidence interval[CI] 12.1–62), smaller tumors (≤T2, OR = 3.6, 95% CI 1.9–7.1), male sex (OR = 2.0, 95% CI 1.1–3.6), overweight/obesity (BMI ≥ 25, OR = 1.9, 95% CI 1.1–3.3), and those married/living with a partner (OR = 1.7, 95% CI 1.0–3.0). In a multivariate model, HPV-positivity (aOR = 27.5, 95% CI 11.4–66) and small tumor size (≤T2, aOR = 2.2, 95% CI 1.0–4.8) remained independently associated with ERα status. When restricted to OPC (n = 180), tumor HPV status (aOR = 17.1, 95% CI 2.1–137) and small tumor size (≤T2, aOR = 4.0 95% CI 1.4–11.3) remained independently associated with ERα expression. ConclusionTumor HPV status and small tumor size are independently associated with ERα expression in HNSCC.

Phytochemicals as Potential Chemopreventive and Chemotherapeutic Agents for Emerging Human Papillomavirus-Driven Head and Neck Cancer: Current Evidence and Future Prospects.

Head and neck cancer (HNC) usually arises from squamous cells of the upper aerodigestive tract that line the mucosal surface in the head and neck region. In India, HNC is common in males, and it is the sixth most common cancer globally. Conventionally, HNC attributes to the use of alcohol or chewing tobacco. Over the past four decades, portions of human papillomavirus (HPV)-positive HNC are increasing at an alarming rate. Identification based on the etiological factors and molecular signatures demonstrates that these neoplastic lesions belong to a distinct category that differs in pathological characteristics and therapeutic response. Slow development in HNC therapeutics has resulted in a low 5-year survival rate in the last two decades. Interestingly, HPV-positive HNC has shown better outcomes following conservative treatments and immunotherapies. This raises demand to have a pre-therapy assessment of HPV status to decide the treatment strategy. Moreover, there is no HPV-specific treatment for HPV-positive HNC patients. Accumulating evidence suggests that phytochemicals are promising leads against HNC and show potential as adjuvants to chemoradiotherapy in HNC. However, only a few of these phytochemicals target HPV. The aim of the present article was to collate data on various leading phytochemicals that have shown promising results in the prevention and treatment of HNC in general and HPV-driven HNC. The review explores the possibility of using these leads against HPV-positive tumors as some of the signaling pathways are common. The review also addresses various challenges in the field that prevent their use in clinical settings.

Is There a Role for Sex Hormone Receptors in Head-and-neck Cancer? Links with HPV Infection and Prognosis.

Head-and-neck squamous cell carcinoma (HNSCC) is the fifth most common cancer in the world and human papillomavirus (HPV) is an important risk factor for this neoplasm. Recent studies showed an association between sex hormone receptors and pathogenesis and/or prognosis in patients with HNSCC. The aim of this study was to clarify the expression patterns of sex hormone receptors in HPV-positive and HPV-negative HNSCC and their associations with tumour biopathology and biological behaviour. Scientific literature indexed in PubMed about sex hormone receptors in HNSCC was retrieved and critically analyzed, to obtain an overview of expression patterns and their possible implications for tumour biopathology and prognosis. Sex hormone receptors were more frequently detected in oropharyngeal tumours compared with HNSCC from other locations. ERα was associated with HPV-positive tumours. The androgen and progesterone receptors were associated with poor patient prognosis. Estrogen receptor alpha (ERα) is implicated in the biopathology of HNSCC in different ways, by promoting DNA hypermutation and facilitating HPV integration thus contributing to an immunogenic phenotype, but also by cooperating with the epithelial growth factor receptor (EGFR) to promote resistance to therapy. The expression of sex hormone receptors may be of prognostic value in specific tumour subgroups, but the use of hormonal therapies for HNSCC is still not in close sight.

Role of High-Risk HPV Detected by PCR in Malignant Sinonasal Inverted Papilloma: A Meta-Analysis.

Prior studies suggest that there may be a link between human papillomavirus (HPV) infection and malignant sinonasal inverted papilloma (SNIP). This systematic review and meta-analysis was performed to further evaluate this potential association. Systematic review with meta-analysis. The Medline and Embase databases were used to identify case-control studies reporting the risk of malignant SNIP in patients with high-risk HPV subtypes identified by polymerase chain reaction (PCR). Meta-analysis was performed to determine pooled odds ratios (ORs) and 95% confidence intervals (CI). Twenty-one studies were identified, including a total of 841 subjects with SNIP. Seventeen studies were included in the final analysis, as four studies did not have any HPV-positive tumors in either group. A total of 56 malignant SNIP and 551 benign SNIP were ultimately identified. The pooled log-OR was 1.80 (95% CI: 1.03-2.57) for all high-risk HPV subtypes. Stratification by high-risk HPV subtype showed a log-OR of 1.67 (95% CI: 0.88-2.46) for HPV-16 and log-OR of 2.68 (95% CI: 1.30-4.05) for HPV-18. Infection with high-risk HPV subtypes may be associated with an increased risk of malignant SNIP. HPV-18 showed the greatest overall average effect size of the common high-risk subtypes. NA Laryngoscope, 132:926-932, 2022.

Abstract 1441: Predictors of treatment response in a preoperative window of opportunity trial of nivolumab in resectable squamous cell carcinoma of the head and neck

Abstract Nivolumab, a PD-1 inhibitor, has variable response rates in patients and is not fully characterized in treatment-naïve patients. Here, we describe exploratory analyses of immune-related gene expression in squamous cell carcinoma of the head and neck (SCCHN) as predictors of response to nivolumab, administered for 4 weeks before planned surgical resection in the context of a window-of-opportunity trial. RNA-seq was performed on pre- and post-treatment tumor tissues from 39 patients with SCCHN of any stage, who were candidates for complete surgical resection (NCT03238365). Expression was quantified using kallisto. Noncoding, histone-coding and mitochondrial transcripts were removed. HPV-specific reads were filtered using pathseq and aligned to HPV reference using viGEN. TME signature scores were quantified using ssGSEA. While HPV-status did not predict clinical responses to nivolumab, the expression of individual HPV16 genes in post-treatment samples reflected response in this patient cohort. Patients who responded to nivolumab showed lower levels of E6 expression, while non-responsive patients had significantly higher E6 expression levels. In contrast, E4 expression was found to be increased in patients who responded to nivolumab treatment. As expected, HPV-positive tumors presented with a more inflamed TME compared to HPV-negative tumors. A tumor microenvironment (TME) classification platform developed by BostonGene was used to analyze several gene expression signatures. Four distinct microenvironment subtypes were reproduced, consistent with pooled analysis. In addition, a high proliferation rate signature in baseline samples was closely associated with non-response to immunotherapy regardless of HPV status. HPV-negative tumors with significantly higher CD8, FoxP3, and CD163 as measured by IHC, had a greater response to immunotherapy in contrast to HPV-positive tumors, where no association was found. This finding was supported by single gene expression and the BostonGene-developed deconvolution algorithm to reconstruct the TME composition of the tumors using RNA sequencing; predicted CD8+ T-cells and Macrophages were significantly enriched in HPV-negative responders. Interestingly, patients with HPV-negative tumors disproportionately responded better to treatment when they presented with an immune-inflamed TME; however, this pattern was not observed in HPV-positive tumors. Our analysis indicates that HPV16 gene transcription appears to impact response to treatment. In addition, the immune-inflamed TME impacts response to nivolumab treatment in HPV-negative tumors. These properties can be assessed in a multiparametric analysis and could lead to the identification of predictors of response to immunotherapy in head and neck cancer. Citation Format: Nikita Kotlov, Zoya Antysheva, Dina Antonova, Naira Samarina, Sandrine Degryse, Alban Linnenbach, Sanket Shukla, Larry Harshyne, Andrew P. South, Jennifer M. Johnson, Young Kim, Alexander Bagaev, Nathan H. Fowler, Adam Luginbuhl. Predictors of treatment response in a preoperative window of opportunity trial of nivolumab in resectable squamous cell carcinoma of the head and neck [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 1441.

Abstract 48: Characterization of Human Papillomavirus (HPV) Sequence and Integration and Analysis of Immune Response Genes for Application in Guatemalan Cervical Cancer

Abstract Purpose: Cervical cancer is a significant cause of mortality in Guatemala due to inefficient screening and late diagnosis. We are developing new technologies to improve the analysis of HPV-positive cervical tumors in Guatemala. The goal is to enhance understanding of HPV infection, tumor progression, and therapeutics development. Methods: We established a cohort of 700 Guatemalan cervical cancer patients with blood, fresh-frozen tumors, and clinical data. To optimize techniques, we extracted DNA and RNA from 20 cervical cancer cell lines. Long-read whole genome and transcriptome sequencing, and long-range PCR, were performed on cervical cancer DNA. HPV and HLA gene sequences were aligned to reference genomes. Results: Complete HPV sequences were compiled, HPV subtypes determined, and integration sites identified in cell lines with single and multiple integration events. HLA types of the class I genes were determined from the complete gene sequence, mutations detected, and HLA homozygosity. RNA was used for long-read whole transcriptome analysis of HPV and cellular genes. These results establish the use of long-read sequencing technology to characterize both HLA and host genomes. Paired Guatemalan tumor and blood DNA was successfully used for full HLA gene sequencing and mutation analysis. In addition, the complete HPV genome sequence was resolved. Binding of HPV peptides was predicted using the HPV and HLA sequences. These peptides identify the specific HPV epitopes potentially presented on the cell surface. Conclusion: Long-read RNA and DNA sequencing technology allow the determination of the complete HPV sequence, integration, and HLA class I typing and mutation analysis. HLA typing provides essential information on an individual's immune response. These techniques and analyses are being applied to tumor samples from women in Guatemala to aid in the development of targeted treatments. These methods are portable and could be employed in low- and middle-income countries. Citation Format: Nicole Rossi, Michael Dean. Characterization of Human Papillomavirus (HPV) Sequence and Integration and Analysis of Immune Response Genes for Application in Guatemalan Cervical Cancer [abstract]. In: Proceedings of the 9th Annual Symposium on Global Cancer Research; Global Cancer Research and Control: Looking Back and Charting a Path Forward; 2021 Mar 10-11. Philadelphia (PA): AACR; Cancer Epidemiol Biomarkers Prev 2021;30(7 Suppl):Abstract nr 48.

Abstract 2205: Molecular subtypes of head and neck cancer in patients with African ancestry

Abstract Epidemiological data identifies disparities in black patients for head and neck squamous carcinoma (HNSCC) screening, detection, treatment, and survival. Although socioeconomic and environmental factors play a role in the existing racial disparities, HNSCC incidence and survival rates in the black population cannot be attributed to these factors alone. To investigate the role race plays in the genomics of head and neck cancer, we examined HNSCC samples from 49 black and 446 white patients in the Cancer Genome Atlas (TCGA). We used computationally-derived ancestry identifications to correlate race with HNSCC molecular features. We then characterized the extent to which genomic alterations in black patients impact their clinical outcomes. We found that black patients lived significantly fewer disease-free months and had a poorer overall survival than white patients. Black patients showed a higher frequency of human papillomavirus (HPV)-negative than HPV-positive tumors compared to white patients. Black patients were found to suffer higher rates of metastasis and more aggressive lymph node involvement. Differences in patient primary tumor site were also identified, with black patients having a higher prevalence of tumors in the larynx and lower prevalence of tumors in the tongue compared to white patients. We also found higher frequencies of RUNX1T1, KMT2D, and UNC13C mutations and lower frequency of PIK3CA mutations in black patients. We found a significant enrichment of MYC signaling and amplifications in the tumors of black patients compared to white patients. Novel data uncovered by this project brings us closer to understanding the racial disparities in HNSCC, and advances research aimed at eliminating those disparities. Unique tumor mutations and molecular signatures show promising clinical utility in providing more targeted, individualized screening diagnostic, and treatment modalities to reduce outcome disparities in the black population. Citation Format: Nadia Mezghani, Alexandria G. Yao, Alison M. Taylor, Fatemeh Momen-Heravi. Molecular subtypes of head and neck cancer in patients with African ancestry [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2021; 2021 Apr 10-15 and May 17-21. Philadelphia (PA): AACR; Cancer Res 2021;81(13_Suppl):Abstract nr 2205.

The unveiled reality of human papillomavirus as risk factor for oral cavity squamous cell carcinoma

The prognostic impact of human papillomavirus (HPV) in oropharyngeal cancer is generally acknowledged, and HPV-status is assessed routinely in clinical practice. Paradoxically, while the oral cavity seems the predilection site for productive HPV-infections, figures on HPV-attribution in oral cavity squamous cell carcinoma (OCSCC) differ widely, and prognostic impact is uncertain. Major obstacles are the lack of reproducible assays to detect HPV in nonoropharyngeal cancers, the relatively small cohorts studied and consequently the shortfall of convincing data. In our study, we used a validated, nucleic acid-based workflow to assess HPV-prevalence in a consecutive cohort of 1016 OCSCCs, and investigated its prognostic impact. In parallel, we analyzed p16-immunohistochemistry (p16-IHC) as surrogate marker for transforming HPV-infection and independent prognosticator. All OCSCC-patients diagnosed between 2008 and 2014 at two Dutch university medical centers were included (N = 1069). Formalin-fixed, paraffin-embedded (FFPE)-samples of 1016 OCSCCs could be retrieved. Punch biopsies were taken from the tumor area in the FFPE-blocks and tested for HPV. P16-IHC was performed on 580 OCSCCs, including all HPV-positive tumors. From 940 samples (92.5%), nucleic acids were of sufficient quality for HPV-testing. In total, 21 (2.2%) OCSCCs were HPV DNA-positive. All HPV DNA-positive tumors were E6 mRNA-positive and considered as true HPV-positive. There was no difference in survival between HPV-positive and HPV-negative OCSCCs. In total, 46 of 580 (7.9%) OCSCCs were p16-immunopositive, including all HPV-positive tumors. Survival was comparable in p16-positive and p16-negative OCSCCs. To conclude, HPV-prevalence is very low in OCSCC and neither HPV-status nor p16-status affects outcome. Based on these data, determining HPV-status in OCSCC seems irrelevant for clinical management.

Prognostic factors for overall survival in oropharyngeal carcinoma depending on HPV status

HintergrundAufgrund der unterschiedlichen Karzinogenese, Prognose und klinischen Manifestation werden seit der 8. Auflage des TNM-Klassifikationssystems der UICC/AJCC (UICC 8) humane Papillomavirus (HPV)-assoziierte und HPV-negative oropharyngeale Plattenepithelkarzinome (OSCC) als 2 Entitäten separat klassifiziert.Material und MethodenInsgesamt 524 Patienten mit einem im Zeitraum von 2000–2016 in der HNO-Klinik des Universitätsklinikums Münster behandelten OSCC wurden hinsichtlich des Nachweises von HPV-Typ-16-spezifischer DNA (HPV16-DNA), des Nikotin- und Alkoholkonsums sowie des Therapieeinflusses auf das Gesamtüberleben (GÜ) untersucht.ErgebnisseEin signifikanter Anstieg der jährlichen Prävalenz der HPV16-DNA-positiven OSCC von 40 % (n = 12/30) im Jahr 2000 auf 46 % (n = 18/39) im Jahr 2016 wurde verzeichnet (p = 0,025, β = 0,539). 89 % (n = 212) der HPV16-DNA-positiven OSCC wurden anhand der UICC 8 gegenüber der UICC 7 herabgestuft. Im Gesamtkollektiv zeigten der häufige Alkohol- und Nikotinkonsum (≥ 10 Packungsjahre) einen statistisch relevanten negativen Einfluss auf das GÜ (p = 0,004 und p = 0,009). Auch häufiger Alkoholkonsum war in der HPV16-DNA-negativen Gruppe prognoserelevant (p = 0,049). In der HPV16-DNA-positiven Gruppe zeigte sich bezüglich des GÜ zwischen den UICC-Stadien I und II (p = 0,481) sowie zwischen III und IV (p = 0,439) gemäß UICC 8 kein statistischer Unterschied.SchlussfolgerungenDie UICC 8 verbessert zwar die Prognosestratifikation der OSCC durch die Trennung von HPV-positiven und HPV-negativen Tumoren im Vergleich zu UICC 7, die prognostische Aussagekraft der UICC 8 für die HPV-assoziierten OSCC ist jedoch weiterhin unzureichend. Der Noxenkonsum könnte zukünftig Einfluss auf die UICC-Klassifikation nehmen, um die prognostische Aussagekraft weiter zu verbessern.

Prognostic significance of human papillomavirus status and treatment modality in hypopharyngeal cancer.

Management of hypopharynx cancer is often extrapolated from larynx cancer. This report analyses treatment patterns and survival limited to hypopharynx cancer using the National Cancer Database (NCDB). There are 9314 patients diagnosed with hypopharynx cancer between 2004 and 2016. The association between treatment modality and survival was analyzed using Kaplan-Meier survival curves and multivariable Cox regression. Five-year overall survival ranged from 45% for stage I to 21% for stage IVB. Treatment modality did not influence survival in stage I/II. For stage III/IV, chemoradiation and surgery + adjuvant therapy were equivalent. Surgery yielded improved survival for T4 disease. Human papillomavirus (HPV)-positive tumors were present in 21% and were associated with improved hazard ratio of death (0.60, p=<0.0001). Survival is superior for T4 hypopharynx cancer managed with surgery, while treatment modality does not impact outcomes for other T-stages. HPV-positive tumors are associated with improved survival regardless of treatment.

NF-Y Subunits Overexpression in HNSCC.

Simple SummaryCancer cells have altered gene expression profiles. This is ultimately elicited by altered structure, expression or binding of transcription factors to regulatory regions of genomes. The CCAAT-binding trimer is a pioneer transcription factor involved in the activation of “cancer” genes. We and others have shown that the regulatory NF-YA subunit is overexpressed in epithelial cancers. Here, we examined large datasets of bulk gene expression profiles, as well as single-cell data, in head and neck squamous cell carcinomas by bioinformatic methods. We partitioned tumors according to molecular subtypes, mutations and positivity for HPV. We came to the conclusion that high levels of the histone-like subunits and the “short” NF-YAs isoform are protective in HPV-positive tumors. On the other hand, high levels of the “long” NF-YAl were found in the recently identified aggressive and metastasis-prone cell population undergoing partial epithelial to mesenchymal transition, p-EMT.NF-Y is the CCAAT-binding trimer formed by the histone fold domain (HFD), NF-YB/NF-YC and NF-YA. The CCAAT box is generally prevalent in promoters of “cancer” genes. We reported the overexpression of NF-YA in BRCA, LUAD and LUSC, and of all subunits in HCC. Altered splicing of NF-YA was found in breast and lung cancer. We analyzed RNA-seq datasets of TCGA and cell lines of head and neck squamous cell carcinomas (HNSCC). We partitioned all TCGA data into four subtypes, deconvoluted single-cell RNA-seq of tumors and derived survival curves. The CCAAT box was enriched in the promoters of overexpressed genes. The “short” NF-YAs was overexpressed in all subtypes and the “long” NF-YAl in Mesenchymal. The HFD subunits are overexpressed, except Basal (NF-YB) and Atypical (NF-YC); NF-YAl is increased in p53 mutated tumors. In HPV-positive tumors, high levels of NF-YAs, p16 and ΔNp63 correlate with better prognosis. Deconvolution of single cell RNA-seq (scRNA-seq) found a correlation of NF-YAl with Cancer Associated Fibroblasts (CAFs) and p-EMT cells, a population endowed with metastatic potential. We conclude that overexpression of HFD subunits and NF-YAs is protective in HPV-positive tumors; expression of NF-YAl is largely confined to mutp53 tumors and malignant p-EMT cells.

A NGS-based Blood Test For the Diagnosis of Invasive HPV-associated Carcinomas with Extensive Viral Genomic Characterization.

Use of circulating tumor DNA (ctDNA) for diagnosis is limited regarding the low number of target molecules in early-stage tumors. Human papillomavirus (HPV)-associated carcinomas represent a privileged model using circulating viral DNA (ctHPV DNA) as a tumor marker. However, the plurality of HPV genotypes represents a challenge. The next-generation sequencing (NGS)-based CaptHPV approach is able to characterize any HPV DNA sequence. To assess the ability of this method to establish the diagnosis of HPV-associated cancer via a blood sample, we analyzed ctHPV DNA in HPV-positive or HPV-negative carcinomas. Patients (135) from France and Senegal with carcinoma developed in the uterine cervix (74), oropharynx (25), oral cavity (19), anus (12), and vulva (5) were prospectively registered. Matched tumor tissue and blood samples (10 mL) were taken before treatment and independently analyzed using the CaptHPV method. HPV prevalence in tumors was 60.0% (81/135; 15 different genotypes). Viral analysis of plasmas compared with tumors was available for 134 patients. In the group of 80 patients with HPV-positive tumors, 77 were also positive in plasma (sensitivity 95.0%); in the group of 54 patients with HPV-negative tumors, one was positive in plasma (specificity 98.1%). In most cases, the complete HPV pattern observed in tumors could be established from the analysis of ctHPV DNA. In patients with carcinoma associated with any HPV genotype, a complete viral genome characterization can be obtained via the analysis of a standard blood sample. This should favor the development of noninvasive diagnostic tests providing the identification of personalized tumor markers. See related commentary by Rostami et al., p. 5158.

Neoadjuvant nivolumab for patients with resectable HPV-positive and HPV-negative squamous cell carcinomas of the head and neck in the CheckMate 358 trial

BackgroundHead and neck squamous cell carcinomas (HNSCCs) are common malignancies caused by carcinogens, including tobacco and alcohol, or infection with human papillomavirus (HPV). Immune checkpoint inhibitors targeting the programmed cell...

Long-term follow-up of bintrafusp alfa, a bifunctional fusion protein targeting TGF-β and PD-L1, in advanced squamous cell carcinoma of the head and neck (SCCHN).

6020 Background: Bintrafusp alfa is a first-in-class bifunctional fusion protein composed of the extracellular domain of the TGF-βRII receptor (a TGF-β “trap”) fused to a human IgG1 mAb blocking PD-L1. A previous report of an expansion cohort from a phase 1 study (NCT02517398) suggested that bintrafusp alfa had a manageable safety profile and early signs of clinical activity in patients with heavily pretreated, advanced SCCHN after a median follow-up of 86.4 weeks. Here we report long-term efficacy and safety for this cohort. Methods: Patients with advanced SCCHN that progressed/recurred after platinum therapy in the recurrent/metastatic setting, or &lt; 6 months after platinum therapy in the locally advanced setting, received bintrafusp alfa 1200 mg every 2 weeks until confirmed progressive disease, unacceptable toxicity, or trial withdrawal. The primary endpoint was confirmed best overall response assessed per RECIST 1.1 assessed by independent review committee (IRC); safety was a secondary endpoint. Results: As of May 15, 2020, 32 patients had received bintrafusp alfa for a median of 2.8 months (range, 0.5-29.9 months), no patient remained on treatment, and median follow-up to data cutoff was 41.7 months (range, 39.8-43.5 months). The objective response rate (ORR; 13%) was unchanged since the previous report; median duration of response (DOR) was increased at 21.4 months (95% CI, 5.5 months to not reached [NR]). While the clinical activity of bintrafusp alfa may be improved in patients with HPV-positive tumors (Table), outcomes were generally similar between PD-L1 subgroups (≥1% vs &lt; 1% tumor cells). The overall safety profile was consistent with the previous report for this cohort, without grade 4 nor 5 treatment-related adverse events (TRAEs); no new TRAEs of grade 3 or that led to discontinuation of bintrafusp alfa were reported. Conclusions: With a median follow-up of over 3 years in patients with heavily pretreated advanced SCCHN, bintrafusp alfa showed sustained clinical activity and 3-year OS of 24.0%, which compares favorably to historical data. Clinical activity appeared to be greater in patients with HPV-positive tumors than those with HPV-negative tumors. The safety profile was manageable and consistent with earlier analysis. Further investigation of bintrafusp alfa in SCCHN and other HPV-associated cancers is ongoing. Clinical trial information: NCT02517398. [Table: see text]

Lack of Conserved miRNA Deregulation in HPV-Induced Squamous Cell Carcinomas.

Squamous cell carcinomas (SCCs) in the anogenital and head and neck regions are associated with high-risk types of human papillomaviruses (HR-HPV). Deregulation of miRNA expression is an important contributor to carcinogenesis. This study aimed to pinpoint commonly and uniquely deregulated miRNAs in cervical, anal, vulvar, and tonsillar tumors of viral or non-viral etiology, searching for a common set of deregulated miRNAs linked to HPV-induced carcinogenesis. RNA was extracted from tumors and nonmalignant tissues from the same locations. The miRNA expression level was determined by next-generation sequencing. Differential expression of miRNAs was calculated, and the patterns of miRNA deregulation were compared between tumors. The total of deregulated miRNAs varied between tumors of different locations by two orders of magnitude, ranging from 1 to 282. The deregulated miRNA pool was largely tumor-specific. In tumors of the same location, a low proportion of miRNAs were exclusively deregulated and no deregulated miRNA was shared by all four types of HPV-positive tumors. The most significant overlap of deregulated miRNAs was found between tumors which differed in location and HPV status (HPV-positive cervical tumors vs. HPV-negative vulvar tumors). Our results imply that HPV infection does not elicit a conserved miRNA deregulation in SCCs.

Daily step counts to predict hospitalizations during chemoradiotherapy for head and neck cancer.

1571 Background: Wearable activity trackers could provide useful data for managing cancer patients with respect to treatment selection, toxicity monitoring, and implementation of supportive care measures. Here, we seek to evaluate the association between daily step counts and hospitalizations in a cohort of patients with head and neck cancer (HNC). Methods: This analysis consists of patients enrolled in one of three prospective trials involving activity monitoring (NCT02649569, NCT03115398, NCT03102229) during chemoradiation. Study subjects were asked to wear a commercial fitness tracker continuously during the therapy. ECOG performance status (PS) was assessed at baseline, and quality of life (QoL) EORTC QLQ-C30 questionnaires were completed weekly. Multivariable Cox regression models with time-dependent covariates (average step count over the past 3 days, most recent QoL score) and time-fixed covariates (age, sex, baseline PS, study number, baseline tumor volume, and treatment setting [definitive versus postoperative]) were used to identify predictors of first hospital admission during the chemoradiotherapy course. In addition to the Cox regression models, linear mixed models were fitted with daily step count as the dependent variable to examine its relationship with certain independent variables including age, sex, weekend status, days after treatment initiation, and study number. Results: Sixty-six HNC patients who received chemoradiotherapy between 2015 and 2019 were included in the analysis. Median age was 60 (range: 27-88). 47% of patients had ECOG PS score 0, 47% ECOG score 1, and 6% ECOG score 2. 29% of patients had HPV-positive oropharyngeal tumors, and the most common other tumor subsites were larynx (27%), and nasopharynx (12%). The Cox regression survival model demonstrated a 26% reduction in the short-term hospitalization risk for every 1000 daily steps (averaged over the past 3 days, hazard ratio 0.74; 95% confidence interval (CI) 0.55-0.98, p = 0.0367). Hospitalizations were not significantly associated with most recent QoL or baseline ECOG PS. Additionally, according to the linear mixed model results, daily step count was not associated with age (p = 0.8048). Study subjects moved less on weekends (on average 245 fewer steps on weekends than weekdays, 95% CI 134-357, p &lt; 0.0001). Also, an increase in most recently measured ECOG PS was associated with a decrease in daily step count (167 fewer steps for every increase in ECOG PS, 95% CI -289 to -45, p &lt; 0.0072). Conclusions: Daily step counts are a dynamic predictor of hospitalizations in patients undergoing chemoradiotherapy for head and neck cancer. Interventional studies are needed to demonstrate feasibility of leveraging physical activity data to optimize supportive care during cancer therapy and enhance cancer care quality. Clinical trial information: NCT02649569, NCT03115398, NCT03102229.

Combined Inhibition of Rad51 and Wee1 Enhances Cell Killing in HNSCC Through Induction of Apoptosis Associated With Excessive DNA Damage and Replication Stress.

Despite advances in surgery, chemotherapy, and radiation, there are limited treatment options for advanced head and neck squamous cell carcinoma (HNSCC) and survival remains very poor. Therefore, effective therapies are desperately needed. Recently, selective exploitation of DNA damage and replication stress responses has become a novel approach for cancer treatment. Wee1 kinase and Rad51 recombinase are two proteins involved in regulating replication stress and homologous recombination repair in cancer cells. In this study, we investigated the combined effect of Rad51 inhibitor (B02) and Wee1 inhibitor (AZD1775) in vitro and in vivo in various HNSCC cell lines. Clonogenic survival assays demonstrated that B02 synergized with AZD1775 in vitro in all HNSCC cell lines tested. The synergy between these drugs was associated with forced CDK1 activation and reduced Chk1 phosphorylation leading to induction of excessive DNA damage and replication stress, culminating in aberrant mitosis and apoptosis. Our results showed that elevated Rad51 mRNA expression correlated with worse survival in HNSCC patients with HPV-positive tumors. The combination of B02 and AZD1775 significantly inhibited tumor growth in vivo in mice bearing HPV-positive HNSCC tumors as compared to HPV-negative HNSCC. This differential sensitivity appears to be linked to HPV-positive tumors having more in vivo endogenous replication stress owing to transformation by E6 and E7 oncogenes. Furthermore, addition of B02 radiosensitized the HPV-negative HNSCC tumors in vitro and in vivo In conclusion, our data implicate that a novel rational combination with Rad51 and Wee1 inhibitors holds promise as synthetic lethal therapy, particularly in high-risk HPV-positive HNSCC.

Molecular landscape of head and neck cancer and implications for therapy.

Head and neck squamous cell carcinomas (HNSCC) arising from the oral cavity, pharynx, and larynx constitute the 6th most common human cancer. Human papillomavirus (HPV)-positive tumours are distinct from HPV-negative counterparts, with HPV status affording clear clinical utility, prognostic benefit and better treatment outcomes. In contrast to their HPV-positive counterparts, HPV-negative tumours are characterized by high mutational load and chromosomal aberrations, with varying copy number alteration (CNA) profiles. HNSCC are distinct tumours at the chromosomal, gene and expression levels, with additional insight gained from immune profiling. Based on mutational analyses, HNSCC are categorized as HPV-positive, HPV-negative CNA-silent, and HPV-negative CNA-high tumours. Furthermore, gene expression profiling segregates these tumours into atypical, classical, basal, and mesenchymal, with clear differences observed between tumours of the oral cavity, oropharynx, hypopharynx and larynx. Additional immune profiling further classifies tumours as either immune-active or immune-exhausted. The clinical utility and impact of these tumour molecular subtypes however remains to be determined. HNSCC harbor high levels of somatic mutations. They display loss at 3p and 18q and gain at 3q and 8q, with mutations in CDKN2A, TP53, CCND1, EGFR, PIK3CA, PTEN, NOTCH1, NSD1, FAT1, AJUBA and KMT2D. Important pathways include the p53 and RB pathways which are involved in cell cycle control and are frequently lost in HPV-negative tumours, the WNT-β-catenin pathway related to the mesenchymal subtype and smoking etiology, and the PI3K pathway which includes the most common genetic alteration in HPV-positive HNSCC. Understanding the mutational, genomic and transcriptomic landscape of HNSCC has leveraged better therapeutic approaches to manage this group of diseases, and it is hoped that additional insight into the molecular subtypes of HNSCC and its specific subsites will further drive improved strategies to stratify and treat patients with this debilitating disease.

P3: CASE-CONTROL STUDY OF SEXUAL BEHAVIOUR IN HPV-POSITIVE ORAL CAVITY AND OROPHARYNGEAL CANCER

Abstract Introduction HPV infection is recognised as the main aetiological factor responsible for the increased incidence of oropharyngeal squamous cell carcinomas (OPSCC). Currently the epidemiology of oral HPV infection and OPSCC is poorly understood. In this study we aim to define the relationship between sexual behaviours and HPV-positive OPSCC. Method We performed a hospital-based, case-control study of 30 patients with HPV-positive and 30 patients with HPV-negative newly diagnosed oral cavity and OPSCC. The participants underwent questionnaire based interviews investigating variables such as age, gender, tobacco use, alcohol intake, and sexual behaviour. Result Subjects in the HPV-positive tumour subgroup had significantly less tobacco use then their HPV-negative counterparts (odds ratio, 0.92, 95% CI 0.9-1.17). A history of oral sexual activity was significantly more likely in the HPV-positive tumour subgroup (odds ratio, 5.23, 95% CI, 1.66-16.51), however this association did not remain significant when adjusted for age, gender, smoking and alcohol consumption. Similarly, there was significantly less alcohol consumption overall in the HPV-positive tumour subgroup (odds ratio, 0.107, 95% CI 0.96-1.01), but this association was no longer significant when adjusted for covariables. Conclusion Significantly less smoking was prevalent in patients with HPV-positive OPSCC. Although patients with HPV-positive tumours had less alcohol consumption and a stronger history of oral sexual activity overall, greater numbers are required in order to show a significant association. This further supports the known association between HPV-infection and OPSCC and the department of health's decision to commence vaccination of boys before the onset of sexual activity. Take-home message HPV-positive oropharyngeal cancer can develop without exposure to established risk factors such as tobacco use or excessive alcohol consumption. This change in pathogenesis should lead a change in how we prevent and manage the disease.