The pursuit of isoform-selective histone deacetylase (HDAC) inhibitors is a promising strategy to overcome the adverse effects associated with pan-HDAC inhibitors. Through systematic structural modification of the known inhibitor PCI-34051, we designed and synthesized two novel series of compounds: 6-substituted indoles and 6-substituted tetrahydroquinolines. Among them, the tetrahydroquinoline derivative 11k exhibited potent HDAC8 inhibitory activity (IC50 = 0.10 μM), and the indole derivative 5b also showed activity (IC50 = 0.28 μM), both with excellent selectivity. Mechanistic studies revealed that these inhibitors specifically increased the acetylation of the HDAC8 substrate SMC3 in cells, without affecting histone H3 or α-tubulin acetylation, supporting their selective on-target action. Preliminary data indicated that compound 11k could modulate cytokine expression in T-cells, highlighting a potential non-cytotoxic biological effect that warrants further investigation. This work introduces novel chemotypes, notably the tetrahydroquinoline scaffold, for the development of selective HDAC8 inhibitors.

Accumulating evidence indicates that N6-methyladenosine (m6A) modification of circular RNAs (circRNAs) plays a pivotal role in regulating cancer progression. However, in head and neck squamous cell carcinoma (HNSCC), the biological functions and underlying mechanisms of m6A modification in circRNAs remain insufficiently elucidated. In this study, we analyzed the association between the expression of IGF2BP3—an upstream m⁶A reader—and clinical outcomes of HNSCC patients, followed by investigating the interaction between IGF2BP3 and circHECTD2 as well as the effect of m⁶A modification on this interaction. Additionally, we explored the molecular mechanism by which IGF2BP3 and circHECTD2 regulate HNSCC progression, focusing on their roles in modulating microRNAs (miRNAs) and target mRNAs, and validated the functional impacts of the IGF2BP3/circHECTD2 axis on HNSCC cell proliferation, invasion, and metastasis. We found high expression of the m⁶A reader IGF2BP3 was significantly associated with poor clinical outcomes in HNSCC patients. Further experiments showed that IGF2BP3 directly bound to circHECTD2 and stabilized it, and m⁶A modification of circHECTD2 enhanced this binding and stabilization effect. Mechanistically, circHECTD2 functioned as a competing endogenous RNA (ceRNA) to sponge hsa-miR-4310 and hsa-miR-7157-5p, thereby preventing the miRNA-mediated degradation of SMAD2 mRNA. Ultimately, the IGF2BP3/circHECTD2/SMAD2 axis was shown to promote HNSCC cell proliferation, invasion, and metastasis. We delineate an m6A-dependent IGF2BP3/circHECTD2/SMAD2 regulatory axis that contributes to HNSCC malignancy. Elevated IGF2BP3 expression correlates with poor patient outcome and enhances circHECTD2 stability through m6A-facilitated binding; circHECTD2 in turn acts as a ceRNA to sequester hsa-miR-4310 and hsa-miR-7157-5p, thereby maintaining SMAD2 expression. Functionally, this axis promotes HNSCC cell proliferation, invasion and metastasis. Collectively, these findings suggest that IGF2BP3 and circHECTD2 may serve as promising prognostic biomarkers and therapeutic targets for HNSCC.

Hypopharyngeal squamous cell carcinoma (HSCC) is an aggressive subtype of head and neck squamous cell carcinoma with insidious onset, early metastasis, and dismal prognosis. Conventional multimodal therapy achieves limited survival benefit, highlighting an urgent need for refined precision strategies. This review systematically summarizes the latest progress in HSCC, covering epidemiological characteristics, imaging diagnosis, prognostic prediction, surgical and non-surgical treatment advances, as well as coding and non-coding therapeutic targets, biomarkers, and candidate drugs. It further introduces the application of multi-omics and machine learning in HSCC research, analyzes current challenges and future directions, and provides a theoretical reference for precise diagnosis, individualized treatment, and prognostic evaluation of HSCC.

Auditory hair cells (HCs) are critical sensory units for sound detection, and their development requires precise transcriptional regulation. To date, numerous transcription factors crucial for HC development have been characterized; however, the governing regulatory network remains unclear and warrants further investigation. In this study, we identified the transcription factor Tox , which plays a significant role in HC development. Tox is highly expressed in HCs in both zebrafish and mice, and its expression increases along developmental pseudotime, rising after Atoh1 and preceding Myo7a , suggesting a potential role in development. Utilizing Tox mutant models in zebrafish and mice, we found that Tox is essential for HC functional maintenance and stereocilia stability. Through multiomics integrated analysis, we confirmed that Tox directly binds to conserved motifs in the Cdh23 promoter, drives its transcription, and ensures the precise localization of Cdh23 at the tips of stereocilia, thereby playing a key role in maintaining auditory function. In conclusion, our findings indicate that Tox influences HC development by regulating the transcription and translation of Cdh23 , enriches the regulatory network of HC development, and may serve as a target for clinical gene therapy for deafness.

Presbycusis, also known as age-related hearing loss (ARHL), is a progressive auditory impairment and ranks among the most prevalent sensory disorders in the elderly population. It is primarily caused by damage to hair cells, degeneration of spiral ganglion neurons, and atrophy of the stria vascularis, which are integral components of the cochlea. While extensive research has been devoted to hair cells and spiral ganglion neurons, this review focuses on the critical role of the stria vascularis in ARHL. The primary function of the stria vascularis is to maintain the endocochlear potential, which is essential for hearing. This review presents an overview of the research concerning the stria vascularis in ARHL, particularly focusing on the application of single-cell transcriptomics to elucidate its role. Furthermore, this review explores relevant signaling pathways and therapeutic strategies for ARHL. By enhancing the understanding of the stria vascularis in ARHL, this review aims to pave the way for personalized treatment and improved strategies for protection and prevention.

Age-related hearing loss (ARHL) is among the most prevalent and complex disorders in older adults. However, the pathogenesis of ARHL remains poorly understood. Using a single-cell transcriptomic landscape of mouse cochlea at five time points (1, 2, 5, 12 and 15 months), we found that the levels of human antigen R (HuR)—a classical RNA-binding protein—increase with age. Here we show that HuR is specifically transported from the nucleus to the cytoplasm in hair cells in both aging mice and nonhuman primates. HuR overexpression in cochlea could successfully alleviate ARHL in aged mice. Meanwhile, HuR deficiency led to premature hearing dysfunction characterized by degeneration of stereocilia and the subsequent loss of hair cells. RNA immunoprecipitation sequencing analysis revealed that HuR can bind to messenger RNAs that enable stereocilia maintenance, including Gnai3. Adeno-associated virus-mediated Gnai3 overexpression partially rescues the hearing defects in HuR-deficient mice. Taken together, these findings indicate that HuR is a potential therapeutic target for ARHL. Age-related hearing loss (ARHL) is a prevalent and complex disorder in older adults. Guo et al. identify human antigen R as a regulator of ARHL in aged mice and demonstrate that cochlear human antigen R overexpression alleviates ARHL, highlighting it as a therapeutic target for the condition.

Due to the absence of effective biomarkers, the precision therapy of head and neck squamous cell carcinoma (HNSCC) still faces challenge. TP53 is one of the most frequently mutated genes in human cancers including HNSCC. Although studies on the regulation of TP53 gene and p53 protein have been extensively explored, the association of TP53-derived circRNAs with HNSCC progression, along with their regulatory mechanisms, remains unknown. This study identifies a novel circRNA derived from TP53 (circTP53), which is upregulated in HNSCC and associated with poor prognosis. It is demonstrated that circTP53 promotes HNSCC progression in vitro and in vivo. Mechanistically, circTP53 interacts with the deubiquitinase USP10, leading to their mutual stabilization, which enhances USP10's deubiquitinating activity on p53, thereby stabilizing p53. Interaction analysis reveals that intron 9 of circTP53 interacts with 100-399AA of USP10. In tumor cells with wild-type p53, circTP53 suppresses cell viability and inhibits the growth of xenograft tumors, while in tumor cells harboring mutant p53, circTP53 demonstrates the opposite effect, enhancing cell viability and promoting xenograft tumor progression. The identification of circTP53 suggests a new direction for p53 research, and the elucidation of circTP53/USP10/p53 axis may provide a new therapeutic scheme for future precision treatment of HNSCC.

Inflammation is among the known causes of cisplatin-induced hearing loss (CIHL), but its exact pathophysiological mechanisms remain unclear. Herein, we demonstrated that pyroptosis-a recently identified inflammatory type of regulated cell death dependent on gasdermin D (GSDMD)-was activated in the cochleae of cisplatin-treated mice, causing CIHL. Meanwhile, treatment with the GSDMD inhibitor necrosulfonamide alleviated CIHL in these mice. To further examine the role of GSDMD-mediated pyroptosis in CIHL, we conducted experiments in Gsdmd-deficient mice. Gsdmd-/- mice demonstrated significantly lower cisplatin-induced cochlear damage than control mice and appeared to be invulnerable to CIHL. Furthermore, GSDMD-mediated pyroptosis in the stria vascularis (SV), but not in the hair cells (HCs), played a dominant role in CIHL. In marginal cells (MCs) of SV, cisplatin induced caspase-dependent GSDMD cleavage, and the pore-forming N-terminal of GSDMD rapidly localized to the mitochondria, leading to abnormal mitochondrial aggregation and oxidative stress. The consequent mitochondrial dysfunction in MCs might result in the severe progression of inflammation, SV damage, and HC loss. Notably, the pharmacological inhibition of pyroptosis using the FDA-approved drug disulfiram effectively alleviated the symptoms of CIHL. Collectively, these findings offer a broad avenue for inhibiting pyroptosis-induced cisplatin ototoxicity and provide valuable theoretical insights for the clinical management of CIHL.

Head and neck squamous cell carcinoma (HNSCC) constitutes a major clinical challenge that severely affects patient survival. Mitochondrial ribosomal protein (MRP) family plays an important role in energy metabolism by participating in mitochondrial oxidative phosphorylation. However, their roles in HNSCC and the underlying mechanisms are still unclear. Single-cell analysis highlighted MRPL21 as a notable biomarker of HNSCC. Human HNSCC tissues, cell lines, and xenograft models in nude mice were used to explore the expression and function of MRPL21. The mass spectrometry was performed to analyze the potential binding targets of MRPL21. In vitro and in vivo experiments were performed to evaluate the effect of MRPL21 on autophagy and cisplatin resistance. The inhibitory actions of siMRPL21 nanodelivery systems on HNSCC progression were also evaluated in vivo. Clinically, relatively high expression level of MRPL21 was associated with poor prognosis in HNSCC patients, and overexpression of MRPL21 significantly promoted HNSCC tumorigenesis, metastasis, and cisplatin resistance. Mechanistically, MRPL21 upregulated mitochondrial oxidative phosphorylation (OXPHOS) and increased PARylation level, inhibited autophagy through activating the downstream PI3K/AKT/mTOR signaling pathway, and ultimately led to tumor progression and cisplatin resistance in HNSCC. We conclude that MRPL21 is a novel biomarker and therapeutic target of HNSCC progression and cisplatin resistant, which may provide a new approach for overcoming cisplatin resistance in HNSCC patients.

Aminoglycoside antibiotic-induced sensorineural hearing loss (SNHL) is a common sensory disorder that requires the development of prophylactic and therapeutic interventions. Lycium barbarum glycopeptide (LBGP) is a peptidoglycan isolated and purified from Lycium barbarum polysaccharides that exhibit significant anti-inflammatory, antioxidant, and neuroprotective effects, but the role of LBGP in aminoglycoside-induced SNHL has not been well investigated. Here it is shown that LBGP can protect against neomycin-induced hearing impairment and alleviate oxidative stress in a neomycin-induced SNHL mouse model. Moreover, it is further found that inhibition of tryptophan hydroxylase (Tph)-mediated serotonin (5-HT) biosynthesis plays a key role in the mechanism of action of LBGP in treating neomycin-induced hearing loss. Systemic delivery of 5-HT increased neomycin-induced apoptosis of cochlear hair cells and spiral ganglion neurons, and pharmacological Tph2 inhibition with P-chlorophenylalanine or Tph2 knock down by AAV-ie-Tph2 effectively attenuated neomycin-induced hearing dysfunction. Collectively, these results provide a promising strategy for the prevention of SNHL by using natural plant extract which is more available and exhibits lower side effects compared with other otoprotective drugs, and identify Tph2 as a potential pharmacological target for the treatment of aminoglycoside-induced ototoxicity.

BACKGROUND:The cellular origin of hypopharyngeal diseases is crucial for further diagnosis and treatment, and the microenvironment in tissues may also be associated with specific cell types at the same time. Normal adjacent tissues (NATs) of hypopharyngeal carcinoma differ from non-tumor-bearing tissues, and can influenced by the tumor. However, the heterogeneity in kinds of disease samples remains little known, and the transcriptomic profile about biological information associated with disease occurrence and clinical outcome contained in it has yet to be fully evaluated. For these reasons, we should quickly investigate the taxonomic and transcriptomic information of NATs in human hypopharynx.RESULTS:Single-cell suspensions of normal adjacent tissues (NATs) of hypopharyngeal carcinoma were obtained and single-cell RNA sequencing (scRNA-seq) was performed. We present scRNA-seq data from 39,315 high-quality cells in the hypopharyngeal from five human donors, nine clusters of normal adjacent human hypopharyngeal cells were presented, including epithelial cells, endothelial cells (ECs), mononuclear phagocyte system cells (MPs), fibroblasts, T cells, plasma cells, B cells, mural cells and mast cells. Nonimmune components in the microenvironment, including epithelial cells, endothelial cells, fibroblasts and the subpopulations of them were performed.CONCLUSIONS:Our data provide a solid basis for the study of single-cell landscape in human normal adjacent hypopharyngeal tissues biology and related diseases.

Background Hypopharyngeal squamous cell carcinoma (HSCC) is a type of head and neck tumor with malignant behavior and poor prognosis. Spatial transcriptomics is a method that spatially analyzes gene expression patterns in tissues and has been used to discover tumor microenvironment and molecular markers in various tumors. However, there are no published reports on spatial transcriptomic analysis of HSCC. Methods In this study, spatial transcriptomic analysis was performed on tumor tissues in situ, peritumoral tissues, and lymphatic metastatic tissues of four patients with HSCC. Morphological markers, including panCK, SMA, and CD45, were used to identify epithelial, fibroblast, and immune cells, respectively. By analyzing the expression of more than 18, 000 genes within the transcriptome of all ROIs, differentially expressed genes of three cell types in different tissues were identified, and differentially expressed signaling pathways and immune infiltration were analyzed. Results The spatial distribution of cells suggests that fibroblast cells in tumor tissues may be involved in the genesis and development of tumors, and the immune infiltration of lymphatic tumor metastasis is lower than that of tumors in situ. For epithelial cells, SLCO2A1, which is a favorable prognosis marker in head and neck squamous cell carcinoma (HNSCC), was significantly down-regulated in tumor tissues and lymphatic metastatic tissues compared with adjacent normal tissues. For immune cells, KANK3, which is a favorable prognosis markers in HNSCC, was significantly down-regulated in lymphatic metastatic tissues compared with adjacent normal tissues. For fibroblast cells, AQP1, CLEC3B and SLCO2A1, which are favorable prognosis markers in HNSCC, were significantly down-regulated in tumor tissues compared with adjacent normal tissues. ITGA8, which is a favorable prognosis markers in HNSCC, was significantly down-regulated in lymphatic metastatic tissues compared with normal lymphatic tissues. CSRP1, DES, and SLCO2A1 positively correlate with immune infiltration in HNSCC. Moreover, SLCO2A1 overexpression suppressed Fadu cells proliferation and metastasis and significantly correlated with favorable survival overcome in HSCC. Conclusions We investigated tumor and fibroblast heterogeneity, as well as the immune microenvironment in HSCC by using spatial transcriptomics. SLCO2A1 may be a tumor suppressor gene and correlates with immune infiltration for HSCC and could serve as a potential target for its diagnosis and treatment.

Bisphenol A (BPA) and its substitute fluorene-9-bisphenol (BHPF) are used in consumer products; however, their toxic effects on intestinal epithelium remain largely unknown. In this study, we combined intestinal organoids and single-cell RNA sequencing to investigate the impact of BPA and BHPF exposure on intestinal cell composition, differentiation, and function. Both compounds inhibited the growth of small intestinal organoids, with BHPF exhibiting a more potent inhibitory effect. BPA and BHPF did not significantly alter the overall cell type composition; however, they led to different alterations in cell-cell communications. Gene Ontology enrichment analysis showed that BPA and BHPF exposures affected various biological processes, such as glutathione transferase activity, antioxidant activity, and lipid metabolism, in cell type-specific and compound-dependent manners. Trajectory analysis demonstrated that BPA and BHPF altered the differentiation trajectory of the intestinal cells. To further connect the cellular mechanism to the phenotypic impact in vivo, , we constructed a mouse model exposed to BPA or BHPF and observed significant alterations in intestinal morphology, including reduced crypt depth and villus length and impaired stem cell proliferation and self-renewal. These results provide novel insights into the cell type-specific effects of BPA and BHPF on the intestinal epithelium and highlight the potential risks of exposure to these compounds. Our findings underscore the importance of evaluating the safety of BPA substitutes and contribute to a better understanding of the effects of environmental chemicals on gut health.

Rationale and Objectives Patients with Hypopharyngeal Squamous Cell Carcinoma (HSCC) exhibiting lymphovascular invasion (LVI) frequently demonstrate a poor prognosis. We aim to determine whether contrast-enhanced computed tomography (CECT)-derived intratumoral and peritumoral radiomic features could predict the LVI status of HSCC patients. Materials and Methods 166 patients with pathologically confirmed HSCC were included in this study, 47 of whom were LVI positive. Preoperative CECT data were randomly divided into a training dataset and a validation dataset in an 8:2 ratio. A total of 1648 radiomics features were extracted from the total tumor volume (GTV) and the surrounding 1- to 5-mm-wide tumor margins (labeled as Peri1V-5V). A deep learning model based on the GTV was also constructed. Radiomics nomograms were established by integrating deep learning model features and clinical features. Receiver operating characteristic curve (ROC), calibration curve, and decision curve analysis (DCA) were utilized to evaluate and compare the predictive performance of all models. Results Peri1V-Radscore showed the best prediction efficiency in the validation dataset among all peritumoral models. Among the clinical variables, the upper tumor boundaries and clinical N stage were independent predictors. Compared with the clinical predictor model, Peri1V-Radscore, deep learn model and Nomogram model can improve prediction efficiency in LVI status. Their respective AUC values were 0.94, 0.84, and 0.96. The results of DCA showed that a good net benefit could be obtained from the Peri1V-Radscore model. Conclusion Intratumoral combined peritumoral radiomics model based on CECT can superior predict LVI status in HSCC patients and may have significant potential for future applications in clinical practice.

Head neck squamous cell carcinoma (HNSCC) is one of the most common malignant tumors which ranks the sixth incidence in the world. Although treatments for HNSCC have improved significantly in recent years, its recurrence rate and mortality rate remain high. Myosin genes have been studied in a variety of tumors, however its role in HNSCC has not been elucidated. GSE58911 and GSE30784 gene expression profile analysis were performed to detect significantly dys-regulated myosin genes in HNSCC. The Cancer Genome Atlas (TCGA) HNSCC database was used to verify the dys-regulated myosin genes and study the relationship between these genes and prognosis in HNSCC. The results showed that MYL1, MYL2, MYL3, MYH2, and MYH7 were down-regulated, while MYH10 was up-regulated in patients with HNSCC. Interestingly, MYL1, MYL2, MYH1, MYH2, and MYH7 were shown to be unfavorable prognostic markers in HNSCC. It is also worth noting that MYL1 was a specific unfavorable prognostic biomarker in HNSCC. MYL1, MYL2, MYL3, MYH2, MYH7, and MYH10 promoted CD4 + T cells activation in HNSCC. MYL1 was proved to be down-regulated in HNSCC tissues compared to normal tissues at protein levels. MYL1 overexpression had no effect on proliferation, but significantly promoted migration of Fadu cells. MYL1 increased EGF and EGFR protein expression levels. Moreover, there is a positive correlation between MYL1 expression and Tcm CD8 cells, Tcm CD4 + cells, NK cells, Mast cells, NKT cells, Tfh cells and Treg cells in HNSCC. Overall, MYL1 facilitates tumor metastasis and correlates with tumor immune infiltration in HNSCC and these effects may be associated with the EGF/EGFR pathway.

The peroxisome is a ubiquitous organelle in rodent cells and plays important roles in a variety of cell types and tissues. It is previously indicated that peroxisomes are associated with auditory function, and patients with peroxisome biogenesis disorders (PBDs) are found to have hearing dysfunction, but the specific role of peroxisomes in hearing remains unclear. In this study, two peroxisome-deficient mouse models (Atoh1-Pex5-/- and Pax2-Pex5-/- ) are established and it is found that peroxisomes mainly function in the hair cells of cochleae. Furthermore, peroxisome deficiency-mediated negative effects on hearing do not involve mitochondrial dysfunction and oxidative damage. Although the mammalian target of rapamycin complex 1 (mTORC1) signaling is shown to function through peroxisomes, no changes are observed in the mTORC1 signaling in Atoh1-Pex5-/- mice when compared to wild-type (WT) mice. However, the expression of large-conductance, voltage-, and Ca2+ -activated K+ (BK) channels is less in Atoh1-Pex5-/- mice as compared to the WT mice, and the administration of activators of BK channels (NS-1619 and NS-11021) restores the auditory function in knockout mice. These results suggest that peroxisomes play an essential role in cochlear hair cells by regulating BK channels. Hence, BK channels appear as the probable target for treating peroxisome-related hearing diseases such as PBDs.

BackgroundHuman hypopharygeal squamous cell carcinoma (HSCC) is a common head and neck cancer with a poor prognosis in advanced stages. The occurrence and development of tumor is the result of mutual influence and co-evolution between tumor cells and tumor microenvironment (TME). Tumor immune microenvironment (TIME) refers to the immune microenvironment surrounding tumor cells. Studying TIME in HSCC could provide new targets and therapeutic strategies for HSCC.MethodsWe performed single-cell RNA sequencing (scRNA-seq) and analysis of hypopharyngeal carcinoma, paracancerous, and lymphoid tissues from five HSCC patients. Subdivide of B cells, T cells, macrophages cells, and monocytes and their distribution in three kinds of tissues as well as marker genes were analyzed. Different genes of IGHG1 plasma cells and SPP1+ macrophages between HSCC tissues, adjacent normal tissues and lymphatic tissues were analyzed. Additionally, we studied proliferating lymphocytes, T cells exhaustion, and T cell receptor (TCR) repertoire in three kinds of tissues.ResultsTranscriptome profiles of 132,869 single cells were obtained and grouped into seven cell clusters, including epithelial cells, lymphocytes, mononuclear phagocytics system (MPs), fibroblasts, endothelial cells (ECs), plasmacytoid dendritic cells (pDCs), and mast cells. Tumor metastasis occurred in three lymphoid tissues. Four distinct populations were identified from lymphocytes, including B cells, plasma cells, T cells and proliferating lymphocytes. We found IGHA1 and IGHG1 specific plasma cells significantly overexpressed in HSCC tissues compared with normal hypopharygeal tissues and lymphatic tissues. Five distinct populations from MPs were identified, including macrophages, monocytes, mature dendritic cells (DCs), Type 1 conventional dendritic cells (cDC1) and Type 2 conventional dendritic cells (cDC2). SPP1+ macrophages were significantly overexpressed in HSCC tissues and lymphatic tissues compared with normal hypopharygeal tissues, which are thought to be M2-type macrophages. Exhaustion of CD8+ Teff cells occurred in HSCC tissues. At last, we verified that IgA and IgG1 protein expression levels were significantly up-regulated in HSCC tissues compared to adjacent normal tissues.ConclusionOverall, this study revealed TIME in HSCC and lymphatic metastasis, and provided potential therapeutic targets for HSCC.

Background:Hypopharyngeal squamous cell cancer (HSCC) is one of the most malignant tumors of the head and neck. It is not easy to detect in the early stage due to its hidden location; thus, lymph node metastasis is highly likely at diagnosis, leading to a poor prognosis. It is believed that epigenetic modification is related to cancer invasion and metastasis. However, the role of m6A-related lncRNA in the tumor microenvironment (TME) of HSCC remains unclear.Methods:The whole transcriptome and methylation sequencing of 5 pairs of HSCC tissues and adjacent tissues were performed to identify the methylation and transcriptome profiles of lncRNAs. The biological significance of lncRNAs differentially expressing the m6A peak was analyzed by Gene Ontology and Kyoto Encyclopedia of Genes and Genomes. By constructing an m6A lncRNA-microRNA network, the mechanism of m6A lncRNAs in HSCC was analyzed. The relative expression levels of selected lncRNAs were examined by quantitative polymerase chain reaction. The CIBERSORT algorithm was used to evaluate the relative proportion of immune cell infiltration in HSCC and paracancerous tissues.Results:Based on an in-depth analysis of the sequencing results, 14413 differentially expressed lncRNAs were revealed, including 7329 up-regulated and 7084 down-regulated lncRNAs. Additionally, 4542 up-methylated and 2253 down-methylated lncRNAs were detected. We demonstrated methylation patterns and gene expression profiles of lncRNAs of HSCC transcriptome. In the intersection analysis of lncRNAs and methylated lncRNAs, 51 lncRNAs with up-regulated transcriptome and methylation and 40 lncRNAs with down-regulated transcriptome and methylation were screened, and significantly differentiated lncRNAs were further studied. In the immune cell infiltration analysis, B cell memory was significantly elevated in cancer tissue, while γδT cell amount was significantly decreased.Conclusion:m6A modification of lncRNAs might be involved in HSCC pathogenesis. Infiltration of immune cells in HSCC might provide a new direction for its treatment. This study provides new insights for exploring the possible HSCC pathogenesis and searching for new potential therapeutic targets.

BACKGROUND:The malignant characteristics of cancer depend not only on intrinsic properties of cancer cells but also on the functions of infiltrating immune cells. In this study, we aimed to investigate the functional landscape of immune cells in head and neck squamous cell carcinoma (HNSCC).METHODS:We employed single-sample gene set enrichment analysis to examine the immunophenotypes of HNSCC based on 29 immune cell functions (ICFs) in TCGA and GSE65858 datasets. We analyzed the clinical features, immune microenvironment, molecular profiles, and biological processes. Additionally, we developed and validated an ICF-based risk score for personalized prognosis prediction. We confirmed the value of the ICF score in our cohort using qRT-PCR and immunohistochemistry. Molecular docking was used to predict potential compounds for immunotherapy.RESULTS:Three immunophenotypes (Immune-L, Immune-M, and Immune-H) were identified in 769 HNSCC samples. The characteristics of Immune-H were consistent with a "Hot" tumor, Immune-L was similar to a "Cold" tumor, and Immune-M exhibited intermediate features. The ICF risk score was associated with immune checkpoints, infiltrating immune cells, tumor mutation burden, and sensitivities to targeted/chemotherapeutic agents. Gene set variation analysis implicated the involvement of metabolic reprogramming pathways in the high-risk group. The combination of "Tumor Immune Dysfunction and Exclusion" and "Immunophenoscore" algorithms indicated that the low-risk group had a higher likelihood of benefiting from immunotherapy. Finally, we identified Eltrombopag and other compounds that may be beneficial for HNSCC immunotherapy.CONCLUSION:Our study provides a novel perspective on the tumor microenvironment of HNSCC, aiding in the understanding of HNSCC heterogeneity and the development of personalized/precision medicine.