The role of SP100 in tumour immune microenvironment and prognosis of head and neck squamous cell carcinoma

BackgroundMetabolic reprogramming is one of the essential features of tumorigenesis. Herein, this study aimed to develop a novel metabolism‐related gene signature for head and neck squamous cell carcinoma (HNSCC) patients.MethodsThe transcriptomic and clinical data of HNSCC samples were collected from The Cancer Genome Atlas (TCGA) and GSE65858 datasets. The metabolism‐related gene‐based prognostic signature (MRGPS) was constructed by the Least Absolute Shrinkage and Selection Operator (LASSO) regression model. The time‐dependent receiver operating characteristic (ROC) and Kaplan‐Meier (K‐M) survival curves were plotted for evaluating its predicting performance. At the same time, univariate along with multivariate analysis was carried out to explore its correlation with clinicopathologic factors. Furthermore, GSEA analysis was performed to explore the signaling pathways affected by MRGPS. We also analyzed the associations of MRGPS with the tumor immune microenvironment (TIME), as well as identified potential compounds via Connectivity Map (CMap) and molecular docking.ResultsA total of 12 differentially expressed metabolism‐related genes were identified and selected to construct the MRGPS. Notably, this signature performed well in predicting HNSCC patients’ survival and could serve as an independent prognostic factor in multiple datasets. In addition to the metabolism‐related pathway, this signature could also affect some immune‐related pathways. The results indicated that MRGPS is correlated with immune cells infiltration and anti‐cancer immune response. Furthermore, we identified cephaeline as a potential therapeutic compound for HNSCC.ConclusionTaken together, we established an MRGs‐based signature that has the potential to predict the clinical outcome and immune microenvironment, which help to search for potential combination immunotherapy compounds and provide a promising therapeutic strategy for treating HNSCC patients.

Ferroptosis and pyroptosis are two new programmed cell death (PCD) modes discovered in recent years. However, the potential value of ferroptosis and pyroptosis-related genes (FPRGs) in prognosis prediction and the tumor immune microenvironment of head and neck squamous cell carcinoma (HNSCC) is still unclear. We obtained 21 significant FPRGs based on the training dataset (TCGA- HNSC) using the univariate Cox and differential expression analysis. The TCGA- HNSC (n = 502) dataset was clustered into two group (clusters A and B) based on the 21 significant FPRGs. 1467 differentially expressed genes (DEGs) between cluster A and B were put into univariate Cox and Least absolute shrinkage and selection operator (LASSO) analysis to build a risk model. The predictive capability of the risk model was successfully confirmed by internal validation, external validation, and clinical sample validation. To improve the clinical applicability, a nomogram model combined risk score and clinical information were constructed. Moreover, the patients with lower risk score were characterized by increased immune response and tumor mutation burden (TMB), while the patients with higher risk score were characterized by increased TP53 mutation rate. In conclusion, our comprehensive analysis of the FPRGs revealed their significant role in prognosis prediction and the tumor immune microenvironment. The risk model containing 9 FPRGs could be a potential prognostic markers and effective immunotherapy targets for HNSCC.

Aging is highly associated with tumor formation and progression. However, little research has explored the association of aging-related lncRNAs (ARLs) with the prognosis and tumor immune microenvironment (TIME) of head and neck squamous cell carcinoma (HNSCC). RNA sequences and clinicopathological data of HNSCC patients and normal subjects were downloaded from The Cancer Genome Atlas. In the training group, we used Pearson correlation, univariate Cox regression, least absolute shrinkage/selection operator regression analyses, and multivariate Cox regression to build a prognostic model. In the test group, we evaluated the model. Multivariate Cox regression was done to screen out independent prognostic factors, with which we constructed a nomogram. Afterward, we demonstrated the predictive value of the risk scores based on the model and the nomogram using time-dependent receiver operating characteristics. Gene set enrichment analysis, immune correlation analysis, and half-maximal inhibitory concentration were also performed to reveal the different landscapes of TIME between risk groups and to predict immuno- and chemo-therapeutic responses. The most important LINC00861 in the model was examined in HNE1, CNE1, and CNE2 nasopharyngeal carcinoma cell lines and transfected into the cell lines CNE1 and CNE2 using the LINC00861-pcDNA3.1 construct plasmid. In addition, CCK-8, Edu, and SA-β-gal staining assays were conducted to test the biofunction of LINC00861 in the CNE1 and CNE2 cells. The signature based on nine ARLs has a good predictive value in survival time, immune infiltration, immune checkpoint expression, and sensitivity to multiple drugs. LINC00861 expression in CNE2 was significantly lower than in the HNE1 and CNE1 cells, and LINC00861 overexpression significantly inhibited the proliferation and increased the senescence of nasopharyngeal carcinoma cell lines. This work built and verified a new prognostic model for HNSCC based on ARLs and mapped the immune landscape in HNSCC. LINC00861 is a protective factor for the development of HNSCC.

ObjectiveTo investigate circadian rhythm-associated long non-coding RNA (lncRNA) signatures in predicting prognosis, metabolism, and immune infiltration in Head and Neck Squamous Cell Carcinoma (HNSC).MethodsHNSC samples were collected from the TCGA database. A signature was constructed using Cox regression and Least Absolute Shrinkage and Selection Operator (LASSO) methods. The immune cell infiltration was analyzed using CIBERSORT, ssGSEA, and MCPcounter. The RT-qPCR was used to detect the expression of signature lncRNAs.ResultsA signature comprising 8 lncRNAs was constructed. The constructed signature demonstrated good prognostic prediction capability for HNSC. A nomogram encompassing risk score accurately predicted the long-term OS probability of HNSC. The infiltration levels of T cell, B cell and Macrophages were significantly higher in the high-risk group than in the low-risk group. Cluster analysis showed that the signature lncRNAs could classify the HNSC samples into two clusters. The RT-qPCR suggested that the expression of lncRNAs in signature was consistent with the data in TCGA.ConclusionThe circadian rhythm-associated lncRNA signature has potential as a prognostic indicator for HNSC. It exhibits associations with metabolism, immune microenvironment, and drug sensitivity, thereby providing valuable insights for informing the treatment of HNSC.

BackgroundDespite the advances in oncology, the prognosis of head and neck squamous cell carcinoma (HNSC) patients remains dismal. The limited response rates to immune checkpoint inhibitors highlight the urgent need for novel therapeutic targets. In this study, we aimed to determine the relevance of PLCXD2 expression in the tumor microenvironment to the HNSC patient clinicopathological features.MethodsGene expression analysis and multicolor immunofluorescence histochemistry with HNSC tissue microarrays were conducted to examine the relation between PLCXD2 expression and patient outcomes. We retrospectively analyzed 275 treatment-naïve patients who underwent surgery for HNSC from 2004–2013. Baseline clinicopathological data, and tumor stage, were retrieved from medical records. The primary prognostic outcome was five-year overall survival, with data censored at the last follow-up for patients who were still alive. Additionally, Spearman correlation analysis was used to assess the relationship between PLCXD2 protein expression and tumor immune infiltrating cells (TIICs), as well as immune checkpoints [programmed cell death protein 1 (PD-1), programmed death ligand 1 (PD-L1) and CTLA-4] in HNSC tissue, while chi-squared test and Cox proportional-hazards models were employed to validate the correlation between PLCXD2 protein levels and clinicopathological characteristics with patient survival.ResultsOur findings revealed higher PLCXD2 protein expression in cancer cells of HNSC tissue compared to control benign tissues (Z=−3.890, P<0.001). Additionally, we observed a distinct association between the presence of PLCXD2 protein in cancer nests and various TIICs, including CD4+ T cells, CD8+ T cells, dendritic cells, as well as CTLA-4+ cells in HNSC tissues. Furthermore, we demonstrated a correlation between PLCXD2 protein expression in cancer cells (hazard ratio: 1.955, P=0.01) and advanced TNM stage (hazard ratio: 1.617, P=0.001), as well as a poorer prognosis.ConclusionsOur findings demonstrate that elevated PLCXD2 expression in HNSC is significantly associated with advanced tumor stage and poorer patient prognosis. The correlation of PLCXD2 with key tumor-infiltrating immune cells and the CTLA-4 checkpoint implicates its role in modulating the tumor immune microenvironment. Therefore, this study supports PLCXD2 as an independent prognostic marker and a potentially promising target for immunotherapy in HNSC.

Background: Immunotherapy has been demonstrated favorable in head and neck squamous cell carcinoma (HNSCC). Studies indicated that immune-related gene prognostic index (IRGPI) was a robust signature, and N6-methyladenosine (m6A) methylation had a significant impact on the tumor immune microenvironment (TIME) and immunotherapy of head and neck squamous cell carcinoma. Thus, combining indicated that immune-related gene prognostic index with m6A status should offer a better predictive power for immune responses. Methods: Head and neck squamous cell carcinoma samples from the cancer genome atlas (TCGA, n = 498) and gene expression omnibus database (GSE65858, n = 270) were used in this study. Cox regression analysis was used to construct the indicated that immune-related gene prognostic index through immune-related hub genes which were identified by weighted gene co-expression network analysis (WGCNA). The m6A risk score was constructed by least absolute shrinkage and selection operator (LASSO) regression analysis. Principal component analysis was used to construct a composite score, and systematically correlate subgroups according to tumor immune microenvironment cell-infiltrating characteristics. Results: A composite score was determined based on indicated that immune-related gene prognostic index and m6A risk score. Head and neck squamous cell carcinoma patients in the cancer genome atlas were divided into four subgroups: A (IRGPI-High&m6A-risk-High, n = 127), B (IRGPI-High&m6A-risk-Low, n = 99), C (IRGPI-Low&m6A-risk-High, n = 99), and D (IRGPI-Low&m6A-risk-Low, n = 128), and overall survival (OS) was significantly different between subgroups (p < 0.001). The characteristics of tumor immune microenvironment cell infiltration in the four subgroups were significantly different in subgroups (p < 0.05). The receiver operating characteristic (ROC) curves show the predictive value of composite score for overall survival was superior to other scores. Conclusion: The composite score is a promising prognostic signature which might distinguish immune and molecular characteristics, predict prognosis, and guide more effective immunotherapeutic strategies for head and neck squamous cell carcinoma.

Squamous cell carcinoma is seen as principal malignancy of head and neck. Tumor immune microenvironment plays a vital role in the occurrence, development and treatment of head and neck squamous cell carcinoma (HNSCC). The effect of immunotherapy, in particular, is closely related to tumor immune microenvironment. This review searched for high-quality literature included within PubMed, Web of Science, and Scopus using the keywords "head and neck cancers," "tumor microenvironment" and "immunotherapy," with the view to summarizing the characteristics of HNSCC immune microenvironment and how various subsets of immune cells promote tumorigenesis. At the same time, based on the favorable prospects of immunotherapy having been shown currently, the study is committed to pinpointing the latest progress of HNSCC immunotherapy, which is of great significance in not only further guiding the diagnosis and treatment of HNSCC, but also conducting its prognostic judgement.

Background: Heat shock protein D1 (HSPD1) is a molecular marker that is significantly highly expressed in numerous malignancies and plays a crucial role in assessing the prognostic status of patients.In the field of head and neck squamous cell carcinoma (HNSCC), the role of the HSPD1 gene in prognostic assessment and its potential link with immune cell infiltration remains largely unexplored, highlighting an urgent need for in-depth scientific research. Methods: In this study, we analyzed the expression data of the HSPD1 gene and its accompanying clinical information from The Cancer Genome Atlas (TCGA) database. The results showed that the expression level of the HSPD1 gene was significantly upregulated in most tumours compared to normal tissues. To validate this observation, we further verified it using Human Protein Atlas data. Through multivariate Cox regression analysis, we found that HSPD1 expression was significantly correlated with several clinicopathological features, suggesting that HSPD1 has the potential to act as an independent factor influencing the survival prognosis of HNSCC patients. Accordingly, we constructed a set of nomogram to more accurately predict the impact of HSPD1 expression on the prognosis of HNSCC patients. Meanwhile, we employed various tools, such as gene ontology analysis, gene set enrichment analysis (GSEA), single-sample GSEA, and the Tumour Immunoassessment Resource database, to explore in depth the biological roles of HSPD1 in HNSCC and its association with immune cell infiltration. Results: The mRNA and protein expression levels of HSPD1 were significantly increased in HNSCC tissues and cell lines. After Cox regression analysis, it was found that HNSCC patients with high HSPD1 expression had shorter overall survival (OS) than those with lower expression in both univariate and multivariate analyses, with statistically significant differences (p-value less than 0.05). In the assessment of the subject's work characteristics (ROC) curve, the area under the curve (AUC) of HSPD1 reached 0.846, showing high predictive accuracy.High expression of HSPD1 was strongly correlated with several clinicopathological features, including pathological N stage, histological grading, lymphovascular invasion, overall survival, and progression-free survival, and there was also a significant association with the patient's smoking history. Further functional enrichment analysis showed that HSPD1 plays an important role in tumourigenesis and cytochrome P450 metabolic pathway. Meanwhile, HSPD1 expression was positively correlated with NK CD56bright, helper T-cells (Th), and Th2 cells; and the infiltration of Mast cells, immature dendritic cells (iDC), Cytotoxic cells, Neutrophils, and mature dendritic cells (DC) was more pronounced in the low-expression group compared with the patients with high HSPD1 expression.Silencing HSPD1 reduced proliferation and migration in SCC9 and Cal27 cell lines. Conclusion: Elevated HSPD1 expression correlates with poor prognosis in HNSCC and impacts tumor immunity. It may function as an oncogene, influencing cell proliferation and migration. The findings highlight the need for in-depth academic research to determine the exact processes and functions.

e18541 Background: Despite considerable advancements in the standard therapies including surgery, chemotherapy and radiotherapy, the clinical outcome for head and neck squamous cell carcinoma (HNSCC) remains poor due to tumor recurrence and metastasis. Molecular-targeted therapies have evolved as novel and promising treatment for HNSCC patients. Herein, we identified a CHK1 inhibitor, prexasertib, as a therapeutic target that enhanced the infiltration of innate and adaptive immune markers in the mice tumor immune microenvironment (TIME) and subsequently sensitizes the tumors. Methods: For this study we used syngeneic mouse model of HNSCC developed using mouse tonsil epithelial cells transformed with HRAS expression and PTPN13 knockdown to represent tobacco-induced HNSCC (MTE-Ras). In vitro drug activity of prexasertib was determined using immunoblotting, flow cytometry, cell proliferation and colony formation assays. The effect of prexasertib on TIME was quantified by Quantigene Plex panel and multiplex immunohistochemistry (mIHC). Results: We found that in vitro treatment with prexasertib increased the amount of DNA damage in the cancer cells and eventually lead to their death. Similarly, our in vivo data showed that treatment with prexasertib resulted in significant tumor regression and increased mice survival. At the molecular level, prexasertib treatment resulted in an upregulation of transcripts associated with T-cell activation, cytokines, chemokines and macrophages indicating an upregulation of inflammatory gene signature. This was further supported by our findings from mIHC staining of mice tumors showing increased infiltration of natural killer cells, natural killer T cells and dendritic cells, following prexasertib treatment. Interestingly, we also saw an increase in the Tregs after prexasertib treatment, which is indicative of an immunosuppressive environment, and we speculate this could be as a result of negative feedback following immune activation by prexasertib treatment. Conclusions: Our results uncover a previously unidentified role of prexasertib in regulating the innate and adaptive immune response in HNSCC, and therefore further corroborate CHK1 as a promising therapeutic target in HNSCC.

BackgroundFunctional interactions between immune cells and neoplastic cells in the tumor immune microenvironment have been actively pursued for both biomarker discovery for patient stratification, as well as therapeutic anti-cancer targets to improve clinical outcomes. Although accumulating evidence indicates that intratumoral infiltration of immune cells has prognostic significance, limited information is available on the spatial infiltration patterns of immune cells within intratumoral regions. This study aimed to understand the intratumoral heterogeneity and spatial distribution of immune cell infiltrates associated with cell phenotypes and prognosis in head and neck squamous cell carcinoma (HNSCC).MethodsA total of 88 specimens of oropharyngeal squamous cell carcinoma, categorized into discovery (n = 38) and validation cohorts (n = 51), were analyzed for immune contexture by multiplexed immunohistochemistry (IHC) and image cytometry-based quantification. Tissue segmentation was performed according to a mathematical morphological approach using neoplastic cell IHC images to dissect intratumoral regions into tumor cell nests versus intratumoral stroma.ResultsTissue segmentation revealed heterogeneity in intratumoral T cells, varying from tumor cell nest-polarized to intratumoral stroma-polarized distributions. Leukocyte composition analysis revealed higher ratios of TH1/TH2 in tumor cell nests with higher percentages of helper T cells, B cells, and CD66b+ granulocytes within intratumoral stroma. A discovery and validation approach revealed a high density of programmed death receptor-1 (PD-1)+ helper T cells in tumor cell nests as a negative prognostic factor for short overall survival. CD163+ tumor-associated macrophages (TAM) provided the strongest correlation with PD-1+ helper T cells, and cases with a high density of PD-1+ helper T cells and CD163+ TAM had a significantly shorter overall survival than other cases.ConclusionThis study reveals the significance of analyzing intratumoral cell nests and reports that an immune microenvironment with a high density of PD-1+ helper T cells in tumoral cell nests is a poor prognostic factor for HNSCC.

Role of Hyaluronan-Mediated CD44 Signaling in Head and Neck Squamous Cell Carcinoma Progression and Chemoresistance

Prognosis for patients with recurrent and/or metastatic head and neck squamous cell carcinoma (HNSCC) remains poor. Development of more effective and less toxic targeted therapies is necessary for HNSCC patients. Checkpoint kinase 1 (CHK1) plays a vital role in cell cycle regulation and is a promising therapeutic target in HNSCC. Prexasertib, a CHK1 inhibitor, induces DNA damage and cell death, however, its effect on the tumor immune microenvironment (TIME) is largely unknown. Therefore, we evaluated a short-term and long-term effects of prexasertib in HNSCC and its TIME. Prexasertib caused increased DNA damage and cell death in vitro and significant tumor regression and improved survival in vivo. The gene expression and multiplex immunohistochemistry (mIHC) analyses of the in vivo tumors demonstrated increased expression of genes that are related to T-cell activation and increased immune cell trafficking, and decreased expression of genes that related to immunosuppression. However, increased expression of genes related to immunosuppression emerged over time suggesting evasion of immune surveillances. These findings in gene expression analyses were confirmed using mIHC which showed differential modulation of TIME in the tumor margins and as well as cores over time. These results suggest that evasion of immune surveillance, at least in part, may contribute to the acquired resistance to prexasertib in HNSCC.

CTLA-4 blockade induces tumor pyroptosis via CD8+ T cells in head and neck squamous cell carcinoma

The phenotype of pyroptosis has been extensively studied in a variety of tumors, but the relationship between pyroptosis and esophageal squamous cell carcinoma (ESCC) remains unclear. Here, 22 pyroptosis genes were downloaded from the website of Gene Set Enrichment Analysis (GSEA), 79 esophageal squamous cell carcinoma samples and GSE53625 containing 179 pairs of esophageal squamous cell carcinoma samples were collected from the Cancer Genome Atlas (TCGA) and the Gene Expression Omnibus (GEO), respectively. Then, pyroptosis subtypes of esophageal squamous cell carcinoma were obtained by cluster analysis according to the expression difference of pyroptosis genes, and a pyroptosis scoring model was constructed by the pyroptosis-related genes screened from different pyroptosis subtypes. Time-dependent receiver operator characteristic (timeROC) curves and the area under the curve (AUC) values were used to evaluate the prognostic predictive accuracy of the pyroptosis scoring model. Kaplan-Meier method with log-rank test were conducted to analyze the impact of the pyroptosis scoring model on overall survival (OS) of patients with esophageal squamous cell carcinoma. Nomogram models and calibration curves were used to further confirm the effect of the pyroptosis scoring model on prognosis. Meanwhile, CIBERSORTx and ESTIMATE algorithm were applied to calculate the influence of the pyroptosis scoring model on esophageal squamous cell carcinoma immune microenvironment. Our findings revealed that the pyroptosis scoring model established by the pyroptosis-related genes was associated with the prognosis and immune microenvironment of esophageal squamous cell carcinoma, which can be used as a biomarker to predict the prognosis and act as a potential target for the treatment of esophageal squamous cell carcinoma.

BackgroundPrevious studies have determined that necroptosis‐related genes are potential biomarkers in head and neck squamous cell carcinoma (HNSCC). Herein, we established a novel risk model based on necroptosis‐related lncRNAs (nrlncRNAs) to predict the prognosis of HNSCC patients.MethodsTranscriptome and related information were obtained from TCGA database, and an nrlncRNA signature was established based on univariate Cox analysis and least absolute shrinkage and selection operator Cox regression. Kaplan–Meier analysis and time‐dependent receiver operating characteristic (ROC) analysis were used to evaluate the model, and a nomogram for survival prediction was established. Gene set enrichment analysis, immune analysis, drug sensitivity analysis, correlation with N6‐methylandenosin (m6A), and tumor stemness analysis were performed. Furthermore, the entire set was divided into two clusters for further discussion.ResultsA novel signature was established with six nrlncRNAs. The areas under the ROC curves (AUCs) for 1‐, 3‐, and 5‐year overall survival (OS) were 0.699, 0.686, and 0.645, respectively. Patients in low‐risk group and cluster 2 had a better prognosis, more immune cell infiltration, higher immune function activity, and higher immune scores; however, patients in high‐risk group and cluster 1 were more sensitive to chemotherapy. Moreover, the risk score had negative correlation with m6A‐related gene expression and tumor stemness.ConclusionAccording to this study, we constructed a novel signature with nrlncRNA pairs to predict the survival of HNSCC patients and guide immunotherapy and chemotherapy. This may possibly promote the development of individualized and precise treatment for HNSCC patients.