Head And Neck Squamous Cancer Research Articles

FAT1 as a tumor mutation burden specific gene affects the immunotherapy effect in head and neck squamous cell cancer

BackgroundResponse to immunotherapy is the main challenge of head and neck squamous cancer (HNSCC) treatment. Previous studies have indicated that tumor mutational burden (TMB) is associated with prognosis, but it is not always a precise index. Hence, investigating specific genetic mutations and tumor microenvironment (TME) changes in TMB-high patients is essential for precision therapy of HNSCC. MethodsA total of 33 HNSCC patients were enrolled in this study. We calculated the TMB score based on next-generation sequencing (NGS) sequencing and grouped these patients based on TMB score. Then, we examined the immune microenvironment of HNSCC using assessments of the bulk transcriptome and the single-cell RNA sequence (scRNA-seq) focusing on the molecular nature of TMB and mutations in HNSCC from our cohort. The association of the mutation pattern and TMB was analyzed in The Cancer Genome Atlas (TCGA) and validated by our cohort. Results33 HNSCC patients were divided into three groups (TMB-low, -medium, and -high) based on TMB score. In the result of 520-gene panel sequencing data, we found that FAT1 and LRP1B mutations were highly prevalent in TMB-high patients. FAT1 mutations are associated with resistance to immunotherapy in HNSCC patients. This involves many metabolism-related pathways like RERE, AIRE, HOMER1, etc. In the scRNA-seq data, regulatory T cells (Tregs), monocytes, and DCs were found mainly enriched in TMB-high samples. ConclusionOur analysis unraveled the FAT1 gene as an assistant predictor when we use TMB as a biomarker of drug resistance in HNSCC. Tregs, monocytes, and dendritic cells (DCs) were found mainly enriched in TMB-high samples.

Dickkopf-1 drives perineural invasion via PI3K-AKT signaling pathway in head and neck squamous cancer.

Perineural invasion (PNI) leads to the poor prognosis of head and neck squamous cancer (HNSCC) patients, but the mechanism of PNI remains unclear. Dickkopf-1 (DKK1), a secretory protein in the Wnt signaling pathway, was found indeed upregulated in HNSCC cells and tissues. Higher expression of DKK1 was statistically relevant to T stage, N stage, PNI, and poor prognosis of HNSCC. DKK1 overexpression enhanced the migration abilities of cancer cells. Moreover, DKK1-overexpressing cancer cells promoted cancer cells invasion of peripheral nerves in vitro and in vivo. Mechanistically, DKK1 could promote the PI3K-AKT signaling pathway. The migration abilities of neuroblastoma cells, which were enhanced by DKK1-overexpressing HNSCC cell lines, could be reversed by an inhibitor of Akt (MK2206). The association of DKK1 with PNI was also confirmed in HNSCC samples. Variables, including T stage, N stage, DKK1 expression, and PNI, were used to establish a nomogram to predict the survival probability and disease-free probability at 3 and 5years. In summary, DKK1 can promote the PI3K-AKT signaling pathway in tumor cells and then could induce neuritogenesis and facilitate PNI. MK2206 may be a potential therapeutic target drug for HNSCC patients with PNI.

Mechanistic insights into the interactions between cancer drivers and the tumour immune microenvironment

BackgroundThe crosstalk between cancer and the tumour immune microenvironment (TIME) has attracted significant interest in the latest years because of its impact on cancer evolution and response to treatment. Despite this, cancer-specific tumour-TIME interactions and their mechanistic insights are still poorly understood.MethodsHere, we compute the significant interactions occurring between cancer-specific genetic drivers and five anti- and pro-tumour TIME features in 32 cancer types using Lasso regularised ordinal regression. Focusing on head and neck squamous cancer (HNSC), we rebuild the functional networks linking specific TIME driver alterations to the TIME state they associate with.ResultsThe 477 TIME drivers that we identify are multifunctional genes whose alterations are selected early in cancer evolution and recur across and within cancer types. Tumour suppressors and oncogenes have an opposite effect on the TIME and the overall anti-tumour TIME driver burden is predictive of response to immunotherapy. TIME driver alterations predict the immune profiles of HNSC molecular subtypes, and perturbations in keratinization, apoptosis and interferon signalling underpin specific driver-TIME interactions.ConclusionsOverall, our study delivers a comprehensive resource of TIME drivers, gives mechanistic insights into their immune-regulatory role, and provides an additional framework for patient prioritisation to immunotherapy. The full list of TIME drivers and associated properties are available at http://www.network-cancer-genes.org.

Somatic 9p24.1 alterations in HPV– head and neck squamous cancer dictate immune microenvironment and anti-PD-1 checkpoint inhibitor activity

Somatic copy number alterations (SCNAs), generally (1) losses containing interferons and interferon-pathway genes, many on chromosome 9p, predict immune-cold, immune checkpoint therapy (ICT)-resistant tumors (2); however, genomic regions mediating these effects are unclear and probably tissue specific. Previously, 9p21.3 loss was found to be an early genetic driver of human papillomavirus-negative (HPV-) head and neck squamous cancer (HNSC), associated with an immune-cold tumor microenvironment (TME) signal, and recent evidence suggested that this TME-cold phenotype was greatly enhanced with 9p21 deletion size, notably encompassing band 9p24.1 (3). Here, we report multi-omic, -threshold and continuous-variable dissection of 9p21 and 9p24 loci (including depth and degree of somatic alteration of each band at each locus, and each gene at each band) and TME of four HPV- HNSC cohorts. Preferential 9p24 deletion, CD8 T-cell immune-cold associations were observed, driven by 9p24.1 loss, and in turn by an essential telomeric regulatory gene element, JAK2-CD274. Surprisingly, same genetic region gains were immune hot. Related 9p21-TME analyses were less evident. Inherent 9p-band-level influences on anti-PD1 ICT survival rates, coincident with TME patterns, were also observed. At a 9p24.1 whole-transcriptome expression threshold of 60th percentile, ICT survival rate exceeded that of lower expression percentiles and of chemotherapy; below this transcript threshold, ICT survival was inferior to chemotherapy, the latter unaffected by 9p24.1 expression level (P-values < 0.01, including in a PD-L1 immunohistochemistry-positive patient subgroup). Whole-exome analyses of 10 solid-tumor types suggest that these 9p-related ICT findings could be relevant to squamous cancers, in which 9p24.1 gain/immune-hot associations exist.

FOXD1 expression in head and neck squamous carcinoma: a study based on TCGA, GEO and meta-analysis

Forkhead box D1 (FOXD1) is a new member of FOX transcription factor family. FOXD1 has demonstrated multilevel roles during normal development, and several diseases’ pathogenesis. However, little is known about the role of FOXD1 in the progression of head and neck squamous cancer (HNSC). In the present study, we analyzed FOXD1 expression pattern using The Cancer Genome Atlas (TCGA) dataset, Gene Expression Omnibus (GEO) datasets, HNSC cell lines, and HNSC tissues. Then, we analyzed the correlation between FOXD1 expression and clinical characteristics, and evaluated the prognostic value of FOXD1 in HNSC. Moreover, we assessed the relationship between FOXD1 expression and tumor microenvironment (TME) and immune cell infiltration using Estimation of STromal and Immune cells in MAlignant Tumor tissues using Expression data (ESTIMATE) and Cell-type Identification By Estimating Relative Subsets Of known RNA Transcripts (CIBERSORT) algorithms. Finally, we predicted the FOXD1-related biological processes (BPs) and signal pathways. FOXD1 was up-regulated in HNSC tissues in TCGA datasets, validated by GEO datasets, HNSC cell lines and HNSC tissues. FOXD1 expression was significantly associated with tumor site and HPV infection. Univariate and multivariate Cox regression analyses showed that FOXD1 expression was an independent prognostic factor. Moreover, we found that the proportions of naïve B cells, plasma cells, and resting dendritic cells (DCs) were negatively correlated with FOXD1 expression, otherwise, the proportion of activated mast cells was positively correlated with FOXD1 expression using CIBERSORT algorithm. Gene Set Enrichment Analyses (GSEAs) revealed that FOXD1 was mainly involved in cancer-related signaling pathway and metabolism-related pathways. FOXD1 was a potential oncogene, and might represent an indicator for predicting overall survival (OS) of HNSC patients. Moreover, many cancer-related pathways and metabolism-related processes may be regulated by FOXD1.

FCGR2A Could Function as a Prognostic Marker and Correlate with Immune Infiltration in Head and Neck Squamous Cell Carcinoma.

Objective We aim to investigate the correlation between FCGR2A mRNA level and prognosis of head and neck squamous cancer (HNSC) in public databases. In addition, we investigated the correlation between FCGR2A expression and clinicopathological characteristics and tumor-infiltrating immune cells in HNSC patients. Methods FCGR2A mRNA expression in multiple cancers was analyzed based on Gene Expression Profiling Interactive Analysis. A protein-protein interaction network was obtained based on the STRING database. The 10 proteins most closely related to FCGR2A (i.e., CD3G, PLCG2, LAT, LYN, SYK, FCGR3A, PIK3R1, HCK, ITGAM, and ITGB2) were screened, followed by establishing the protein-protein interaction network. The correlation between FCGR2A expression and immunocytes was investigated, together with the effects of FCGR2A on the metastasis, recurrence, and survival of HNSC. Results FCGR2A expression in several carcinoma tissues was significantly higher than that of adjacent tissues. Significant differences were noticed in the HNSC samples and the adjacent tissue samples except the seven samples of grade 4. There were statistical differences between the FCGR2A expression in tissues of grade 1, grade 2, and grade 3 (P < 0.05). In the tissues of grade 4, the expression of FCGR2A was the lowest. The FCGR2A protein was a type of II-a receptor in γFc of the low-affinity immunoglobulin, which could bind with the Fc region of the immunoglobulin γ. There was a correlation between the FCGR2A gene and the distal HNSC metastasis. FCGR2A gene expression was correlated with the survival and prognosis. The GSE65858 dataset was selected for the validation. The FCGR2A expression was significantly correlated with total survival (P = 0.0107) and progression-free survival (P = 0.0362). Conclusions Our findings shed light on the importance of FCGR2A in HNSC and illustrated a potential relationship between FCGR2A and tumor-immune interactions.

Osteopontin Levels in Patients with Squamous Metastatic Head and Neck Cancer.

Background and Objectives: Increased osteopontin (OPN) concentrations in the plasma of patients with head and neck squamous cancer (HNSCC) have diagnostic significance, and it can indicate more aggressive biological behavior of cancer. The aim of this study was to determine OPN levels in patients with HNSCC of different primary locations and to assess its prognostic significance in metastasis development. Materials and Methods: This cohort study included 45 patients (41 male and 4 female patients) with HNSCC with different primary localization of head and neck. All patients underwent surgery—neck dissection. All patients were categorized according to the histological findings of the resected material and tumor–node–metastasis (TNM) classification system. After surgery, N categories were determined on the basis of histological features of resected material. Results: The histological findings of our patients showed: N0 in 11 patients, N1 in 8 patients, N2a in 4 patients, N2b in 14 patients and N2c in 8 patients. Plasma OPN values in all study participants ranged from 2.24 to 109.10 ng/mL. OPN levels in plasma of patients with negative nodes compared to the group of patients with positive nodes in the neck differed significantly (16.89 ng/mL to 34.08 ng/mL, respectively; p = 0.03). There were significantly lower OPN plasma levels in the group of subjects with histologically positive one lymph node in the neck (N1) compared to the group of patients with N2b histologically positive findings of resected neck material (10.4 ng/mL to 43.9 ng/mL, respectively; p = 0.02). Conclusions: The results have shown that growing N degrees of positive neck nodes classification were accompanied by growing values of plasma osteopontin. Osteopontin might be important for the development of neck metastases.

Comprehensive analysis of macrophage-related multigene signature in the tumor microenvironment of head and neck squamous cancer.

Macrophages are among the most abundant cells of the tumor microenvironment in head and neck squamous cancer (HNSC). Although the marker gene sets of macrophages have been found, the mechanism by which they affect macrophages and whether they further predict the clinical outcome is unclear. In this study, a univariate COX analysis and a random forest algorithm were used to construct a prognostic model. Differential expression of the key gene, methylation status, function, and signaling pathways were further analyzed. We cross-analyzed multiple databases to detect the relationship between the most critical gene and the infiltration of multiple immune cells, as well as its impact on the prognosis of pan-cancer. FANCE is recognized as hub gene by different algorithms. It was overexpressed in HNSC, and high expression was predictive of better prognosis. It might promote apoptosis through the Wnt/β-catenin pathway. The expression of FANCE is inversely proportional to the infiltration of CD4 + T cells and their subsets, tumor-associated macrophages (TAMs), M2 macrophages, but positively co-expressed with M1 macrophages. In summary, FANCE was identified as the hub gene from the macrophage marker gene set, and it may improve the prognosis of HNSC patients by inhibiting lymphocytes and tumor-associated macrophages infiltration.

Clinical data analysis reveals the role of OGR1 (GPR68) in head and neck squamous cancer.

BackgroundHead and neck squamous cancer (HNSC) frequently occurs in the clinic. Revealing the role of the genes that correlate with cancer cell outgrowth will contribute to potential treatment target identification and tumor inhibition.MethodsThe gene expression profiles and gene ontology of the proton‐sensing G‐protein‐coupled receptor OGR1 were analyzed using the TCGA (The Cancer Genome Atlas) database. The effects of sex, age, race, and degree of malignancy on HNSC were investigated, and the survival times of HNSC patients with high or low/medium expression levels of OGR1 were compared. Methylation of the OGR1 promoter CpG sites was also investigated and OGR1‐related genes were analyzed using gene set enrichment analysis.ResultsOGR1 is overexpressed in HNSC patients. However, compared with the low/median expression group, the high OGR1 expression group did not have different survival rates. The OGR1 expression level differed across sex, age, race, and degree of malignancy, while the methylation of the OGR1 promoter CpG sites was maintained at a similar level. Gene set enrichment analysis revealed that OGR1 was positively correlated with head and neck cancer, cisplatin resistance, hypoxia, angiogenesis, cell migration, and TGF‐β.ConclusionThe expression of OGR1 correlated with HNSC progression and survival and thus can serve as a potential treatment target and prognostic marker.

Multicomponent Analysis in Head and Neck Squamous Cancer: Based on Genome, Epigenome and Transcriptome

In recent years, with the rapid development and application of a series of high-throughput proteomics technologies such as gene chip, high-throughput sequencing, and mass spectrometry, people have a more comprehensive and systematic understanding of diseases (including tumors) at genome, transcriptome, proteome and metabolome levels. The research papers on genome sequencing and gene expression profiling of head and neck tumors are increasing year by year, which provides a lot of basic data for revealing the pathogenesis of tumors and excavating new biomarkers. Although there are some articles on the joint analysis of multi-genomics, most of them are based on the speculation of coding amino acid mutations based on gene expression patterns and gene mutations. In this study, our analysis revealed four distinct head and neck squamous cancer (HNSC) subtypes through genome, epigenome and transcriptome data, which provides the possibility for molecular pathological diagnosis, subtyping, invasion and metastasis, chemoradiotherapy resistance and tumor heterogeneity of head and neck tumors. Funding: The work was supported by The Natural Science Youth Founds of Jiangxi Province (no.2016BAB215249), The Natural Science Foundation of Fujian Province (no. 2015J01512), The Middle- and Young-Aged Key Personnel Projects of Fujian Provincial Health and Family Planning Commission, China (no. 2015-ZQN-JC-39). Declaration of Interest: The authors declare that they have no competing interests. Ethical Approval: It is considered by the Ethic Committee of the Quanzhou First Hospital Affiliated to Fujian Medical University, The experimental design and scheme of this study fully considered the principle of safety and fairness, this study does not cause harm or risk to the study subjects. The subjects will be based on the principle of voluntariness, the right to know is guaranteed, protect the interests and privacy of research subjects to the maximum extent, There is no conflict of interest between the research results and the research content. Ethics Committee agrees the program to carry out as planned (No. [2019] 109).

The FSHR Expression in Head and Neck Squamous Cell Cancer. A Pilot Immunohistochemical Study.

Follicle-stimulating hormone receptor (FSHr), expressed on endothelial cells of vessels in different malignant tumors, has been recently investigated as a potential pan-receptor of cancer treatment. However, the expression of this receptor has also been confirmed in other tissues under pathological conditions including cancer. The aim of the presented pilot study was to evaluate the expression of FSHr in head and neck squamous cancer (HNSCC). A total of 28 HNSCC patient samples were immunohistochemically analyzed for the presence of FSHr using a commercially available primary antibody. FSHr was detected not only in the tumor tissue, but also in the basal layer or dysplastic parts of squamous mucosa and also in fibroblasts surrounding the tumor tissue. FSHr is present on different benign or malignant mesenchymal and epithelial structures in HNSCC. A brief literature review revealed a wider role of FSHr in the development of neoplasia.

Incidence and risk factors of interstitial lung disease of patients with head and neck cancer treated with cetuximab.

Interstitial lung disease (ILD) is known as a potentially severe adverse event associated with epidermal growth factor receptor (EGFR)-targeted therapy. The incidence and risk factors of ILD in patients with head and neck squamous cancer (HNSCC) treated with cetuximab, an anti-EGFR monoclonal antibody, have not been established. We retrospectively reviewed patients with HNSCC who received cetuximab from December 2012 to December 2016 at our institute and evaluated the incidence and risk factors of ILD. Of the 201 patients with HNSCC, ILD was observed in 9 patients (4.5%), 8 of whom had grade 3 or higher. High Krebs von den Lungen-6 (KL-6) and ≥50 pack-years of smoking were significantly predictive of associated with ILD (P = 0.00011 and 0.05, respectively). The incidence of ILD in patients with HNSCC treated with cetuximab was <5%, but most of the ILD cases were severe. High KL-6 and smoking histories might be predictive for ILD among patients with HNSCC.

A phase I trial of T4 CAR T-cell immunotherapy in head and neck squamous cancer (HNSCC).

3046Background: Recent FDA approvals make CAR T-cell therapy a clinical reality for hematologic malignancy. Toxicity and antigen loss-mediated resistance remain problematic. Solid tumors impose add...

Longitudinal analyses of CD3ζ-chain expression in the correlation of the disease status in a cohort of head and neck cancer patients (TUM2P.1033)

Abstract Purpose: Despite advances in multimodality treatment, the 5-year survival rate of Head and neck squamous cancer (HNSCC) patients has unimproved over the last 4 decades. CD3ζ has emerged as a clinically important immunological molecule in HNSCC. Its relevance as a prognostic biomarker of HNSCC, however, has not been formally addressed in a longitudinal study. Methods: Peripheral blood mononuclear cells (PBMC) were collected from 46 patients and 53 controls at the time of diagnosis and post treatments (upto 2 year period). Expression of CD3ζ in the T cells of the PBMC samples were analyzed in flow cytometry. Results: We standardized a method for longitudinal analyses of intracellular CD3z expressions in the PBMC samples. We considered a &amp;lt;10% baseline increase in the normalized MFI of CD3ζ expression as a predictor of disease status in evaluating the follow-up samples of the HNSCC patients. Correlation analysis showed that 27/29 HNSCC patients who showed an increase in the CD3ζ expression relative to their baselines were disease free (negative predictive value. 93.1%). 10/17 HNSCC patients who showed a reduced/no change in the CD3ζ expression died or had recurrent disease (positive predictive value, 58.8 %). Overall accuracy of the assay was 80.43%. The sensitivity and specificity were respectively 83.3% and 79.41%. Conclusion: Our longitudinal analyses supported that a &amp;gt;10% increase in the CD3ζ expression against baseline could be a good prognostic biomarker in HNSCC patients.

Abstract 450: Head and neck cancer and HPV infection: A potential role for peritumoral neurogenesis in HPV-associated malignancy

Abstract Background: Head and neck cancers account for 5% of all cancer related deaths and present with complex molecular pathology and clinical features owing to their functional anatomical location. The molecular etiology of head and neck squamous cancer (HNSC) has gained extensive attention recently revealing key mutations, viral infection, and an emerging landscape of varying disease sub-types. While perineural invasion has long been associated with poor outcome in HNSC, the association of neurogenesis-related genes with cancer progression has only recently come into focus in HNSC. Using prospectively acquired patient samples and TCGA HNSC mutational analyses we further characterized the sequelae of molecular changes in neurogenesis-related genes and propose that these genes play a significant role in HNSC development. Methods: All clinical specimens were collected under approval by Institutional Review Board. Prospectively collected samples were analyzed for HPV DNA by linear arrays (InnoLipa) and q-PCR for E7-RNA. Gene expression arrays were conducted to determine differences in HPV activity and neuronal associated gene ontologies (Agilent). Gene expression profiles were compared in oral HPV+ and HPV- cultured oral keratinocytes as well as the TCGA Kandoth &amp; Tang datasets. Gene targets were validated by Immunohistochemistry and qPCR. Results: Initial unsupervised hierarchical clustering analysis of our HNSC cohort revealed a separation of HPV+ HPV+E7-(HPV-inactive: HPVin), and HPV- tumors with changes in the expression of key neurogenesis-related genes. For example, POU4F1, NGFR, and GRIN2C were significantly up-regulated (P&amp;lt;0.05) in HPV+ vs HPVin cancers. Additional analysis of TCGA data revealed divergence of the mutational spectra in HNSC with and without perineural invasion. Conclusion: Our preliminary findings indicate the emerging importance of neurogenesis-related genes and perineural involvement in HNSC and support a divergence of HPV+, HPVin, and HPV- HNSC sub-types. Citation Format: Christian A. Graves, Swati Tomar, Diego Altomare, James R. Wells, Kim E. Creek, Lucia Pirisi. Head and neck cancer and HPV infection: A potential role for peritumoral neurogenesis in HPV-associated malignancy. [abstract]. In: Proceedings of the 105th Annual Meeting of the American Association for Cancer Research; 2014 Apr 5-9; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2014;74(19 Suppl):Abstract nr 450. doi:10.1158/1538-7445.AM2014-450

Photothermal Ablation of Head and Neck Squamous Cell Carcinoma by Nanoparticle Loaded Macrophages

Objectives: Investigate the effect of photothermal treatment with gold nanoshells (NS) using macrophage-mediated delivery system in head and neck squamous cancer (HNSC) cell line. Methods: The human HNSC cell line (FaDU) and Rat Ma (NR8383) was used for both monolayers and spheroids experiments. Ma were loaded with gold nanoshells. In spheroid models, the cytotoxicity of FaDU cell spheroid with NS loaded Ma was compared to FaDU cell spheroid with empty Ma (without NS). Monolayers and spheroids were exposed to increasing levels of 810nm laser light. The effect of laser ablation treatment was evaluated using MTS assay and spheroid growth kinetics. Live/dead assay was examined by two photon microscopy. Results: The cell viability (MTS assay) of PTT-treated FaDU/NS loaded MS monolayers decreased in a laser dose-dependent and NS loaded MS cell count. In live/dead assay, PTT-treated hybrid FaDU spheroid with NS loaded Ma show significant cell death after exposure to NIR laser light. In kinetics of spheroid growth, PTT-treated hybrid FaDU spheroids containing NS loaded Ma displayed significant decrease in their growth pattern compared to that observed for spheroids containing empty Ma (without NS). Conclusions: NIR laser irradiation of HNSC monolayers or spheroids incorporating NS loaded Ma resulted in significant growth inhibition in a irradiance dependent manner. With further development, photothermal treatment with gold nanoparticle loaded macrophages have the potential to improve the quality of life of head and neck cancer patients, providing minimally invasive modality for cancer treatment.

Seamless phase II/III trial design with survival and PRO endpoints for treatment selection: Case study of RTOG 1216.

TPS6099 Background: Clinical trial results from phase II trials to select an experimental treatment arm for separate phase III trial comparison can require years. Cancer clinical trials also now aim at both survival and PRO/functional outcomes, especially in head and neck (HN) studies. We developed a unique seamless phase II/III trial design to save on sample size and trial duration. The initial multi-arm phase II trial selects the most effective regimen among multiple experimental arms by first comparing each of the new treatments to a common control arm, using chosen endpoints, such as progression free survival. The winner will be tested for overall survival in the phase III study. Methods: We propose a phase II/III design to test the efficacy of experimental arms of postoperative radiation (RT) + docetaxel or RT + docetaxel + cetuximab in patients with HN squamous cancer. These are compared to the control arm of RT + cisplatin in the phase II part. Only one arm will be selected to go on to phase III depending on efficacy (PFS), PRO and safety outcomes. One experimental arm must be sufficiently better than the common control arm and the winner not having increased toxicity or functional cost to be selected for phase III inclusion. If not, the trial is halted for futility. Patients in the phase II selected arm and the control arm are included in phase III testing. Group sequential method is used to design each component. Separate interim efficacy and futility analyses are built in such that each endpoint can be monitored as in separate phase II, III trials. Once sample sizes are derived, operating characteristics for the seamless II/III design are evaluated through simulations under the null and various alternative hypotheses. Savings on sample size and time are compared to typical separate phase II and III designs and to the design testing only the arm of RT + docetaxel + cetuximab in phase II. Conclusion: The phase II/III RTOG 1216 HNC trial offers cost effectiveness, operational efficiency and scientific innovation.

Abstract 4709: EGFR inhibition augments anti-tumor immune responses by HER family-specific human CD4 helper T cells in vitro.

Abstract Despite immunotherapy is a promising approach for cancer treatment, there are many obstacles remain to be overcome. The characteristics of tumor evasion of immune surveillance are produced by decreased expression of immune molecules and secretion of immunoregulatory cytokines. To break these tumor microenvironment which act as suppressor of antitumor immunology, we evaluated the value of EGFR inhibitors as immune-modulators. Following two days EGFR tyrosine kinase inhibitor treatment, expression of HLA-DR molecule of head and neck squamous cancer (HNSCC) cell lines were increased, meanwhile EGFR blocking antibody treatment augmented HLA-DR expression. Furthermore, EGFR inhibitors decreased spontaneous secretion of IL-8 and TGF-β from HNSCC cell lines. To confirm the immune supporting activity of EGFR inhibitors, tumor antigen specific CD4 helper T lymphocyte (HTL) responses to EGFR-expressing tumor were investigated. We identified novel EGFR HTL epitope using MHC class II binding algorithms. The candidate EGFR peptide elicited HTL responses against EGFR-expressing HNSCC in vitro. The EGFR peptide shares high homology to HER2, HER-3 and c-Met, so that these were able to stimulate EGFR peptide reactive HTLs responses in a HLA-DR restricted manner indicating that the EGFR peptide serves a universal tumor-associated peptide in HER family and c-Met. Finally, we evaluated EGFR reactive HTL responses against HNSCC cell lines treated with EGFR inhibitors. As a result, EGFR inhibitors drastically increased HTL IFN-γ production and cytotoxic response. Since HER family and c-MET have been considered as targets of alternative pathway of EGFR inhibition, therapeutic strategy combining EGFR inhibitors with novel HTL epitope identified in this study might be attractive option for antitumor immunotherapy. Citation Format: Takumi Kumai, Yasuaki Harabuchi, Hiroya Kobayashi. EGFR inhibition augments anti-tumor immune responses by HER family-specific human CD4 helper T cells in vitro. [abstract]. In: Proceedings of the 104th Annual Meeting of the American Association for Cancer Research; 2013 Apr 6-10; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2013;73(8 Suppl):Abstract nr 4709. doi:10.1158/1538-7445.AM2013-4709

63IN - Implications for Clinical Practice and Trial Design

ABSTRACT Targeted strategies have been developed in head and neck squamous cancer (HNSCC) in order to increase effectiveness and decrease toxicity. Epidermal growth factor receptor (EGFR) inhibitors are firmly established in HNSCC but the critical issue has been patient selection. Other recently developed novel pathway inhibitors hold significant promise and are being tested in phase 1, 2 and 3 trials either alone or concomitantly with radiotherapy or chemoradiotherapy. One such class of agents, the poly(ADP-ribose) polymerase (PARP) inhibitors, has shown activity in conjunction with radiotherapy in head and neck cancer cell lines. Pre-clinical data suggest that PARP inhibitors may potentiate the effects of radiotherapy in head and neck cancer cell lines and xenograft models. Sonic hedgehog (Shh) pathway plays an important role in HNSCC progression and should be considered a potential therapeutic target. One clinical arena where targeted therapies may also find place is in the adjuvant or prevention setting where minimal disease or precancerous lesions can be affected by shutdown of a single or few targets. Future directions lie in understanding the molecular basis of the mechanism of action of the drugs currently undergoing clinical evaluation. Increased understanding of the cell signaling processes involved might aid patient selection. Advances in genomics are revolutionizing the discovery of biomarkers and should improve the tumor selectivity of novel agents. There is also a need for the development of methods that can be used in pharmacodynamic studies in humans. A better understanding of the signaling pathways will not only inform our knowledge of cancer development but also help to refine the classification as well as the treatment of the disease. Disclosure The author has declared no conflicts of interest.

Management of Head and Neck Cancer: Surgical and Nonsurgical

AbstractIt is right time to review the management of head and neck squamous cancer (HNSC) because of fundamental changes in both diagnostic and therapeutic modalities. Head and neck cancer affects area highly associated with the individual's.identity and can produce profound alteration in appearance, speech, and swallowing. Due to morbidity, disfigurement and problems of disease control clinicians used to have lot of reservations in treating complex HNSC cases. The field has taken a new vigor by incorporating important basic advances in understanding of cancer, new modalities of treatment and management of functional deficits. Although much progress has been made in understanding the molecular genetics of HNSC for novel diagnostic and therapeutic interventions, they are still far to go before becoming standard of care for head and neck cancer.