The detection of circulating tumor cells (CTCs) using immunoaffinity-based methods often relies on epithelial-related markers, which may bias the selection of CTCs and limit the biological information obtained, depending on the targeted antigens. Herein, we compared the molecular profiles and clinical significance of CTCs based on the expression of epithelial-related markers (EPCAM, EGFR, and MET) in patients with head and neck squamous-cell carcinoma (HNSCC). CTCs were detected using density gradient separation and CD45-negative selection, followed by quantitative PCR for epithelial-related marker expression. Expression profiles of epithelial–mesenchymal transition (EMT)-related (VIM, CDH1, CDH2, SNAI1, ZEB1, ZEB2, and TWIST1) and immune-regulatory (CD274 and PDCD1LG2) genes were compared. Moreover, the association between marker expression and clinical factors was analyzed. Among the 60 patients with CTCs, 48 (80.0%), 20 (33.3%), and 31 (51.7%) were positive for EPCAM, EGFR, and MET, respectively. A significant correlation was observed between CTCs expressing EPCAM and EGFR. CTCs expressing distinct markers showed differing EMT-related and immune-regulatory gene expression. EPCAM+ CTCs were associated with advanced-stage disease, while EGFR+ CTCs were correlated with locoregional relapse and shorter progression-free survival (p = 0.007; hazard ratio = 3.254). Patients with EPCAM/EGFR double-positive CTCs had the poorest prognosis. These findings emphasize the importance of marker selection in liquid biopsy technologies and highlight the need for improved detection methods and the further investigation of CTC biology.

BACKGROUND:Like tumor cells, cancer-associated fibroblasts (CAFs) can enter the bloodstream and may function in systemic circulation. METHODS:Circulating CAFs (cCAFs) were isolated from the peripheral blood of head and neck squamous cell carcinoma (HNSCC) patients using CD45 depletion and detected by FAP expression using RT-qPCR. Circulating tumor cells (CTCs) were detected through the expression of epithelial markers (EPCAM, EGFR, and MET). We analyzed the association between cCAF positivity and clinical factors, CTC characteristics, and verified tumor tissue relationships using public datasets. RESULTS:Of the 97 patients, 19.6% were positive for cCAFs and 61.9% for CTCs. cCAF positivity was significantly associated with lymph node metastasis, CTC positivity, and lower EPCAM levels on CTCs. In tumor tissues, FAP expression inversely correlated with EPCAM expression in tumor cells. CONCLUSION:cCAFs may contribute to the survival of CTCs and tumor progression through changes in the characteristics of CTCs.

T cell memory is an important mechanism for long-term protection against diverse pathogens such as cancers. Tissue-resident memory T cell (TRM) is a subset of memory T cells that stably resides in tissues and controls local immune homeostasis. In the present study, we investigated the immunological profile and clinical significance of TRM in patients with head and neck squamous cell carcinoma (HNSCC). We analyzed RNA-sequencing data and clinical data obtained from The Cancer Genome Atlas (TCGA) database. In total, 520 cases, including 97 human papillomavirus (HPV)-positive HNSCCs and 423 HPV-negative HNSCCs, were analyzed. TRM-enriched tumors were identified based on the expression of both CD4/CD8A and ITGAE (CD103). TRM-enriched tumors tended to correlate with HPV-positive status and early T factor. Gene Set Enrichment Analysis revealed the enrichment of pathways relating to cytotoxic immune response in TRM-enriched tumors. Moreover, the expressions of immunostimulatory and immune checkpoint-related genes were elevated in TRM-enriched tumors. Notably, the enrichment of CD4+TRMs in tumors was an independent prognostic factor for shorter progression-free survivals (PFS). We also analyzed peripheral blood mononuclear cells obtained from 77 patients with HNSCC. The increased CD8+CD103+TRMs correlated with HPV-positive status and primary lesion of oropharynx. The proportion of CD4+CD103+TRMs negatively correlated with that of CD4+ naïve T cells, while that of CD8+CD103+TRMs negatively correlated with that of effector T cells. The proportion of CD4+CD103+TRMs positively correlated with Ki-67 expression in CD4+ T cells. Moreover, the elevated CD4+CD103+TRMs was an independent prognostic factor for shorter PFS. In conclusion, we highlighted the clinical and immunological significance of CD103+TRMs in both tumor tissues and peripheral blood of patients with HNSCC. Further characterization of TRMs could lead to the development of novel biomarkers for patients with HNSCC. Hideyuki Takahashi, Shota Ida, Hiroe Tada, Kazuaki Chikamatsu. CD4+CD103+ tissue-resident memory T cells correlate with unfavorable prognosis in patients with head and neck cancer [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 1 (Regular Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_1):Abstract nr 6031.

BACKGROUND/AIM:Human papillomavirus (HPV)-positive head and neck squamous cell carcinoma (HNSCC) is clinically and immunologically distinct from HPV-negative HNSCC. Herein, we investigated the presence of tumor antigens HPV E6/E7 and wild-type p53-specific T-cell responses, and the impact of immune checkpoint blockade in patients with HPV-positive HNSCC. MATERIALS AND METHODS:Peripheral blood mononuclear cells (PBMCs) from patients with HPV-positive HNSCC were stimulated with HPV E6/E7 or wild-type p53-derived peptide mixture and evaluated using the interferon-γ enzyme-linked immunosorbent spot assay. Flow cytometry was performed to analyze the proportion of T-cell subsets and T cells expressing immune checkpoint molecules. RESULTS:HPV E6/E7-specific T cells were detected in 22 (95.7%) of 23 patients, whereas wild-type p53-specific T cells were detected in 3 (15.0%) of 20 patients. Seven (43.8%) of 16 patients exhibited wild-type p53-specific T-cell responses, as determined using whole proteins instead of peptides. Immune checkpoint blockade enhanced wild-type p53-specific T-cell responses in 9 (45.0%) of 20 patients. Flow cytometric analysis of PBMCs revealed that responders exhibiting enhanced wild-type p53-specific T-cell responses following immune checkpoint blockade had a significantly higher proportion of Ki-67+CD4+ T cells, Ki-67+CD8+ T cells, regulatory T cells, PD-1+CD4+ T cells, and TIM-3+CD4+ T cells than non-responders. CONCLUSION:Our findings indicate that tumor antigen-specific T cells are present in the peripheral blood of patients with HPV-positive HNSCC. Blockade of checkpoint pathways can enhance T-cell responses in certain patients, probably via activated T cells, Tregs, and/or exhausted CD4+ T cells.

Abstract Innate lymphoid cells (ILCs) and natural killer (NK) cells are a heterogeneous family of innate immune cells that regulate inflammation, immune tolerance and tissue homeostasis in infection, chronic inflammation, metabolic diseases and cancer. The ILC family lacks the expression of antigen-specific receptors and demonstrate effector function by generating cytokines and secreted proteins. In tumor microenvironment, ILCs play a dual role including both tumor regression and tumor promotion, suggesting their diverse phenotypes and functional states. Among the ILC family, ILC1s and NK cells share some features, including the expression of the transcription factor T-bet and the ability to produce IFN-γ. In the present study, we analyzed single-cell RNA sequencing (scRNAseq) data to investigate the landscape and functional status of ILC families in patients with head and neck squamous cell carcinoma (HNSCC). The GSE164690 dataset, including pre-processed scRNAseq data and clinical data, was obtained from Gene Expression Omnibus. A total of 17 pairs of tumor-derived CD45-positive cells and peripheral blood lymphocytes, consisting of 6 HPV-positive HNSCCs and 11 HPV-negative HNSCCs, were analyzed using the Seurat v4 R package. After quality control, a total of 95,809 cells were obtained and clustered into 16 immune cell clusters. Next, a total of 7,278 NK cells were extracted and sub-clustered into 11 clusters. To characterize the 11 clusters, the abundances of NK subsets were estimated by CIBERSORTx. This approach identified eight NK cell clusters, two intraepithelial ILC1 (ieILC1) clusters, and one ieILC1-NK-intermediate (ieILC1-NK-int) cluster. Single-sample GSEA (ssGSEA) analysis was performed to further characterize each subset. ieILC1 clusters expressed tissue-resident markers ITGA1, CD69, and CXCR6, whereas ieILC1-NK-int cluster expressed ILC1 markers CCR7, IL7R, and SELL. Principle component analysis (PCA) based on ssGSEA indicated that ieILC1 clusters and ieILC1-NK-int showed the different PCA distribution from NK clusters. Inflammation and effector function-related pathways, including inflammatory response, interferon-gamma response, and interferon-alpha response, were enriched in ieILC1 clusters compared with the others. Among ieILC1 clusters, several pathways, including hypoxia, TNFA signaling via NFKB, allograft rejection, TGF beta signaling, were more enriched in ieILC1-1 than in ieILC1-2, suggesting that ieILC1-1 is the most immunologically active phenotype among NK subclusters. Of note, ieILC1-2 mainly comprised cells isolated from HPV-negative tumors, whereas ieILC1-1 mainly comprised cells isolated from HPV-positive tumors. In conclusion, we have demonstrated the diverse landscape of NK and ieILC1 cells in patients with HNSCC. Further characterization of the ILC family would lead to a novel therapeutic approach against HNSCC. Citation Format: Hideyuki Takahashi, Shota Ida, Hiroe Tada, Kazuaki Chikamatsu. Activated phenotype of intraepithelial type 1 innate lymphoid cells is enriched in HPV-positive head and neck cancers [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2024; Part 1 (Regular Abstracts); 2024 Apr 5-10; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2024;84(6_Suppl):Abstract nr 2685.

Carbon‐ion radiotherapy (C‐ion RT) is effective for head and neck mucosal melanoma (HN‐MM), including radioresistant mucosal melanoma. Melanoma also responds effectively to immune checkpoint inhibitors (ICIs). Data on the efficacy and safety of ICIs for HN‐MM are insufficient.

OBJECTIVES:Exosome-mediated reciprocal crosstalk between tumor and stromal cells plays a crucial role in tumor development and progression. This study investigated whether exosomes released from head and neck squamous cell carcinoma (HNSCC) tumor cells can convert normal fibroblasts into cancer-associated fibroblasts (CAF)-like cells and further analyzed the functional characterization of fibroblasts educated by tumor-derived exosomes.MATERIALS AND METHODS:Exosomes secreted from HNSCC cell lines were isolated and normal fibroblasts were established from normal oropharyngeal mucosa. The effects of the exosomes on fibroblasts were examined by proliferation and migration assays, and exosome-educated fibroblasts were analyzed for the expression of eight genes (IL1B, IL6, CXCL8, TGFB1, ACTA2, FAP, CD274, and PDCD1LG2) by RT-qPCR. Moreover, T cells or CD14-positive cells were co-cultured with culture supernatants from exosome-educated fibroblasts. T-cell proliferation and macrophage polarization were examined using flow cytometry. Then, RNA sequencing (RNA-seq) of exosome-educated fibroblasts and the corresponding control fibroblasts was performed.RESULTS:Tumor-derived exosomes enhanced fibroblast proliferation and migration. Moreover, gene expression analysis revealed upregulation of the gene expression of proinflammatory cytokines and immunoregulatory genes, and activated fibroblast marker genes. The culture supernatants of tumor-derived exosome-educated fibroblasts suppressed T cell proliferation and the induction of protumoral macrophages compared with those of control fibroblasts. Next, comprehensive RNA-seq analysis data revealed the activation of 11 signaling pathways, including IL-6- and IL-17-related signaling.CONCLUSION:These results indicate that HNSCC tumor cells induce and/or differentiate into CAFs through exosome-based cell-to-cell communication to create an inflammatory tumor microenvironment.

Objectives: Human papillomavirus (HPV)-positive head and neck squamous cell carcinoma (HNSCC) differs in terms of cellular and molecular biological characteristics from HPV-negative HNSCC. However, differences in circulating tumor cells (CTCs) between HPV-positive and -negative HNSCC remain unclear.Materials and methods: We first analyzed eight epithelial-mesenchymal transition (EMT)-related genes (VIM, CDH1, CDH2, SNAI1, SNAI2, TWIST1, ZEB1, and ZEB2) using The Cancer Genome Atlas (TCGA) database. Next, we isolated CTCs from patients with HNSCC using CD45-negative selection and expression analysis of epithelialrelated genes (EPCAM, EGFR, and MET) by RT-qPCR. CTC-positive samples were further analyzed for EMTrelated genes. In addition, we investigated the proportion of circulating T cell subsets and CD38+ T cells using flow cytometry and their involvement in CTCs.Results: Compared with HPV-negative HNSCC, expression of CDH1, SNAI1, SNAI2, TWIST1, and ZEB1 was downregulated in HPV-positive HNSCC, as determined by TCGA analysis. CTCs were detected in 19 (52.8 %) of 36 HPV-positive and 26 (68.4 %) of 38 HPV-negative patients with HNSCC. EPCAM-positive and MET-positive CTCs were significantly more frequent in patients with HPV-negative HNSCC. HPV-positive patients with HNSCC exhibited significantly high SNAI1 and ZEB2 expression in CTCs. Interestingly, differences in SNAI1 expression levels differed markedly between CTCs and TCGA based on HPV status. Moreover, HPV-positive patients with HNSCC exhibiting SNAI1-high CTCs showed a superior prognosis and a lower proportion of CD38+ T cells than those with SNAI1-low CTCs.Conclusion: Our results provide novel insights into the EMT-MET spectrum of CTCs and may contribute to the development of prognostic biomarkers for HPV-positive HNSCC.

of salivary gland malignant tumor and has been reported to be a mammary analogue secretory carcinoma due to its morphological resemblance to secretory carcinoma of the breast. SC is newly listed in the WHO Classification of Head and Neck Tumors （4th edition） as a slow-growing and low-grade malignant tumor of the salivary gland. SC may have been diagnosed as acinic cell carcinoma or adenocarcinoma not otherwise specified. Herein, we present four cases of SC regarding histopathological and immunochemical findings and an expression of specific ETV6-NTRK3 fusion genes. Of the four cases, the submandibular and parotid glands were involved in one and three cases, respectively. All patients were male, and the age ranged from 46 years to 63 years. There were no cases of facial palsy before treatment and no specific findings on the sonogram or magnetic resonance imaging scan. SC was suspected in Case 1 from preoperative fine-needle aspiration cytology with immunochemical staining by Pan-Trk and NOR-1. Malignancy was suspected in Case 4 although the histological type was inconclusive. Surgery was performed in all cases. In two patients （Cases 1 and 4）, postoperative radiation was performed due to the close surgical margin. The diagnosis of SC was made based on histopathological findings, immunochemical staining （mammaglobin, S-100, Pan-Trk, and NOR-1）, and the presence of ETV6-NTRK3 fusion genes. In three cases （Cases 2, 3, and 4）, the ETV6-NTRK fusion gene was detected by the RT-PCR or FISH method. In Case 1, detection of fusion gene was not performed because of the immunochemical positivity of Pan-Trk which has a high specificity for NTRK3 fusion, and negativity of NOR-1 for acinic cell carcinoma. The post-treatment course was uneventful, and Case 4 was well without recurrence or metastasis for 5 years. fusion gene, Pan-Trk immunochemis-try, creation of multiple slides from a single smear preparation for cytology, malignant salivary gland tumor

Background/Aim: Bone and soft-tissue sarcomas of the head and neck have very poor prognoses. This prospective study aimed to investigate the efficacy and safety of carbon-ion radiotherapy (C-ion RT) for bone and soft-tissue sarcoma of the head and neck. Patients and Methods: The present study was a prospective clinical study that included 10 consecutive patients diagnosed with bone and soft-tissue sarcoma of the head and neck who were treated with C-ion RT between 2012 and 2018 at our institution. C-Ion RT consisted of 70.4 Gy (relative biological effectiveness) in 16 fractions. Results: The 3-year local control, overall survival, and progression-free survival rates for patients overall were 72.9%, 77.8%, and 36%, respectively. Conclusion: The present study demonstrated the efficacy of C-ion RT for bone and soft-tissue sarcoma of the head and neck; adverse events were within the expected range.

External auditory canal (EAC) cancer is a rare disease for which there are no adequate evidence-based treatment strategies. Radiotherapy is often used as the initial treatment to preserve the organ. This study aimed to elucidate the efficacy of radiotherapy for EAC squamous cell carcinoma (SCC). Patients with T1 disease were treated with radiotherapy alone. Patients with T2–4 disease were treated with chemoradiotherapy. The median follow-up period was 30.4 months. The 3-year local control (LC) rate for all patients was 51%, the disease-free survival (DFS) rate was 44%, and the overall survival (OS) rate was 73%. For T1–3 disease, the 3-year LC rate was 74%, DFS was 62%, and OS was 89%. However, for T4 disease, the 3-year LC rate was 17%, DFS was 17%, and OS was 50%. In a univariate analysis, only the T-category was a significant factor for LC and DFS (p = 0.006 and 0.02, respectively). All local recurrences were within the high-dose irradiated area. The results of this study suggest chemoradiotherapy can be an alternative to a combination of surgery and postoperative radiation for T1–3 SCC of the EAC. However, the efficacy of chemoradiotherapy in T4 cases was inadequate.

Objective: Data on the long-term outcomes of carbon-ion radiotherapy for non-squamous cell carcinomas are rare. Gunma University has been utilizing carbon-ion radiotherapy as a treatment for head and neck cancer since 2010. This study aimed to evaluate the long-term efficacy and safety of carbon-ion radiotherapy for the head and neck. Methods: We prospectively evaluated 35 patients with non-squamous cell carcinoma who underwent carbon-ion radiotherapy at Gunma University Heavy Ion Medical Center between 2010 and 2014. The 5-year local control, overall survival, and progression-free survival rates were evaluated. Results: The median age was 59 years (range, 31-77 years), and the median follow-up time was 65 months (range, 6.1-98.8 months). Overall, 32 and 3 patients received 64.0 Gy (relative biological effectiveness) and 57.6 Gy (relative biological effectiveness) in 16 fractions, respectively. Adenoid cystic carcinoma was the most dominant histopathological type (n = 21 patients, 60 %). The 5-year local control, progression-free survival, and overall survival rates were 74.5 %, 53.2 %, and 81.3 %, respectively. Nine patients had local recurrence, and six patients died. Adverse events included acute grade 3 radiation mucositis (Common Terminology Criteria for Adverse Events version 4.0) in 8 patients; this was improved immediately with conservative therapy. Late grade 4 adverse events were observed, including two cases of visual loss and one case of brain necrosis. No grade 5 adverse events were observed. Conclusion: Carbon-ion radiotherapy achieves excellent local control and overall survival rates in non-squamous cell carcinoma patients with expected adverse events. (C) 2021 Asian AOMS, ASOMP, JSOP, JSOMS, JSOM, and JAMI. Published by Elsevier Ltd.

T-cell memory is an important mechanism for long-term protection against diverse pathogens. Generation and persistence of memory T cells are vital components of anti-tumor immunity, given their ability to persist for prolonged durations, as well as activate and migrate rapidly. In the present study, we investigated the clinical and prognostic significance of T-cell subsets in the peripheral circulation of patients with head and neck squamous cell carcinoma (HNSCC). Moreover, we calculated the enrichment scores of T-cell subsets in primary tumor tissues and compared their clinical characteristics using a public database. Multivariate survival analyses of circulating T-cell parameters revealed that clinical parameters, except M factor, were not independent prognostic factors, whereas proportions of CD8+ T cells, naïve T cells (TN s), effector memory T cells (TEM s), and CD38+ CD8+ T cells were independent prognostic factors, suggesting the importance of these peripheral T-cell parameters as independent prognostic biomarkers. Consistent with these results, the T-cell enrichment analysis indicated that enrichment of CD8+ TN s in the tumor microenvironment was an independent prognostic factor. Moreover, an ex vivo experiment demonstrated significantly less cytotoxic activity in CD38+ T cells than in CD38- T cells. These findings suggest that T-cell memory-related parameters in both systemic immunity and the tumor microenvironment could be used as prognostic biomarkers regardless of clinical characteristics. Further characterization of circulating T cells would lead to the development of novel biomarkers for patients with HNSCC.

Head and neck squamous carcinoma (HNSCC) is highly infiltrated by immune cells, including tumor-infiltrating lymphocytes and myeloid lineage cells. In the tumor microenvironment, tumor cells orchestrate a highly immunosuppressive microenvironment by secreting immunosuppressive mediators, expressing immune checkpoint ligands, and downregulating human leukocyte antigen expression. In the present study, we aimed to comprehensively profile the immune microenvironment of HNSCC using gene expression data obtained from public database. We calculated enrichment scores of 33 immune cell types based on gene expression data of HNSCC tissues and adjacent non-cancer tissues. Based on these scores, we performed non-supervised clustering and identified three immune signatures—cold, lymphocyte, and myeloid/dendritic cell (DC)—based on the clustering results. We then compared the clinical and biological features of the three signatures. Among HNSCC and non-cancer tissues, human papillomavirus (HPV)-positive HNSCCs exhibited the highest scores in various immune cell types, including CD4+ T cells, CD8+ T cells, B cells, plasma cells, basophils, and their subpopulations. Among the three immune signatures, the proportions of HPV-positive tumors, oropharyngeal cancers, early T tumors, and N factor positive cases were significantly higher in the lymphocyte signature than in other signatures. Among the three signatures, the lymphocyte signature showed the longest overall survival (OS), especially in HPV-positive patients, whereas the myeloid/DC signature demonstrated the shortest OS in these patients. Gene set enrichment analysis revealed the upregulation of several pathways related to inflammatory and proinflammatory responses in the lymphocyte signature. The expression of PRF1 , IFNG , GZMB , CXCL9 , CXCL10 , PDCD1 , LAG3 , CTLA4 , HAVCR2 , and TIGIT was the highest in the lymphocyte signature. Meanwhile, the expression of PD-1 ligand genes CD274 and PDCD1LG2 was highest in the myeloid/DC signature. Herein, our findings revealed the transcriptomic landscape of the immune microenvironment that closely reflects the clinical and biological significance of HNSCC, indicating that molecular profiling of the immune microenvironment can be employed to develop novel biomarkers and precision immunotherapies for HNSCC.

We report a case of diffuse large B-cell lymphoma (DLBCL) of the pterygopalatine fossa (PPF) that developed with multiple cranial nerve palsies. A 70-year-old woman received steroid treatment for right facial palsy but was referred to our hospital because of additional diplopia and dysarthria. Neurological examination revealed multiple cranial nerve palsies associated with the right IV and XII and bilateral VII nerves. Computed tomography and magnetic resonance imaging showed lesions infiltrating Meckel’s cavity and the cavernous sinus from the right PPF. Subsequently, right visual impairment and III and V1 cranial nerve palsies appeared; however, the right XII and left VII cranial nerve palsies improved spontaneously. A biopsy from the PPF was performed using an endoscopic transnasal approach, and the histological diagnosis was DLBCL. She was treated with chemotherapy. Although the PPF is an anatomically difficult site to access, biopsy using the endoscopic transnasal approach makes a minimally invasive diagnosis possible.

Objectives: Tumor-infiltrating T cell (TIL) is a major cell type involved in tumor eradication in the tumor microenvimnment (TME). Among TILs, tissue-resident memory T cells (T(RM)s) have been recognized as a subset capable of continuous immunosurveillance to afford long-term immunity. In the present study, we comprehensively profiled T-RM in patients with head and neck squamous cell carcinoma (HNSCC). Materials and Methods: We analyzed RNA-sequencing (RNA-seq) data obtained from The Cancer Genome Atlas (TCGA) database. Based on the gene expression of CD69 and CD4/CD8A, we identified T-RM-enriched patients and evaluated their clinical and biological significance. In addition, we analyzed peripheral blood mononuclear cells (PBMCs) obtained from 60 patients with HNSCC to evaluate the presence of T-RM-like cells in the peripheral circulation. Results: TCGA analysis revealed that T-RM-enriched tumors correlated with early T factor, human papillomavirus-positive status, the proportion of oropharynx lesion, upregulated inflammatory pathways, upregulation of immunostimulatory and immune checkpoint molecule genes, and favorable overall survival. Moreover, we clarified the presence of CD69 + T-RM-like cells that highly express PD-1 and TIM-3 in the peripheral circulation of patients with HNSCC. Conclusion: We highlighted the clinical and transcriptomic significance of T-RM in patients with HNSCC. Further characterization of T-RM could lead to the development of novel biomarkers, especially for immune checkpoint therapies.

The evaluation of antitumor immune responses is essential for immune monitoring to predict clinical outcomes as well as treatment efficacies in cancer patients. In this study, we produced two tumor antigen (TA) proteins, melanoma antigen family A4 and wild type p53, using TG silkworm systems and evaluated anti-TA-specific immune responses by enzyme-linked immunosorbent spot assays in patients with head and neck cancer. Eleven (61.1%) of 18 patients showed significant IFN-γ production in response to at least one TA; however, the presence of TA-specific immune responses did not significantly contribute to better prognosis (overall survival, p = 0.1768; progression-free survival, p = 0.4507). Further studies will need to be performed on a larger scale to better assess the clinical significance of these systems. The production of multiple TA proteins may provide new avenues for the development of immunotherapeutic strategies to stimulate a potent and specific immune response against tumor cells as well as precise assessment of antitumor immune responses in cancer patients.

Altered metabolism is an emerging hallmark of cancer. Cancer cells preferentially utilize glycolysis for energy production, termed “aerobic glycolysis.” In this study, we performed a comprehensive analysis of the glycolysis status in the tumor microenvironment (TME) of head and neck squamous cell carcinoma (HNSCC) using data from The Cancer Genome Atlas database. We first divided 520 patients with HNSCC into two groups based on the mRNA expression of 16 glycolysis-related genes. The glycolysis-high signature positively correlated with human papillomavirus-negative tumor type, advanced T factor, and unfavorable prognosis. The gene set enrichment analysis revealed upregulation of several pathways, including interferon-alpha response, myc targets, hypoxia, epithelial-mesenchymal transition, transforming growth factor-β signaling, and interleukin 6-Janus kinase-signal transducer and activator of transcription 3 signaling, in the glycolysis-high group. Immune cell enrichment analysis revealed decreased infiltration of T cells, dendritic cells, and B cells in the glycolysis-high group, suggesting impaired tumor antigen presentation, T cell activation, and antibody production in TME. Moreover, the expression of TGFB1 , CD274 , and PDCD1LG2 , which facilitate immunosuppression in the TME, was upregulated in the glycolysis-high group. Collectively, these findings suggest the potential of glycolysis monitoring as a biomarker for tumor progression and immunosuppression in the TME of HNSCC.