Abstract Background: Tumor-related leukocytosis (TRL), driven largely by neutrophil overproduction, is associated with aggressive disease, treatment resistance, and poor survival across solid tumors. In metastatic urothelial carcinoma (mUC), patients with TRL experience particularly poor outcomes under immune checkpoint inhibitor (ICI) therapy, suggesting an immune-suppressive tumor microenvironment. However, the molecular basis linking TRL to ICI resistance remains unclear. Methods: We retrospectively evaluated mUC patients receiving ICIs to assess the prognostic role of TRL. RNA from 35 tumors underwent NanoString profiling to identify leukocytosis-associated genes. IL13RA2 and related candidates were validated across UC cell lines by qPCR, western blotting, and functional assays including proliferation, migration, cytokine analysis, and neutrophil chemotaxis. Transcriptional programs regulated by IL13RA2 were defined using Clariom S microarray. Results: Patients with baseline leukocytosis (WBC >10,000/µL) showed significantly worse overall survival, confirming TRL as a negative prognostic factor during ICI therapy. High-WBC tumors displayed a distinct inflammatory signature, with IL13RA2 among the most upregulated genes. IL13RA2 expression strongly associated with neutrophil enrichment in TCGA BLCA (Pearson r = 0.41; TIMER ρ = 0.199) and was validated in the ICI-treated GSE176307 cohort, where IL13RA2-high tumors exhibited increased neutrophil and myeloid fractions. Functionally, IL13RA2 knockdown reduced proliferation, migration, and wound-healing capacity in UMUC3 and T24 cells. Microarray analysis identified reproducible IL13RA2-regulated gene sets enriched for cytokine, inflammatory, motility, and chemotaxis pathways. Conditioned media from IL13RA2-expressing cells robustly induced migration of purified neutrophils, whereas IL13RA2 depletion markedly impaired neutrophil chemotaxis. IL13RA2 silencing suppressed JAK1/2-STAT3 activity and reduced multiple pro-inflammatory cytokines and chemokines, including CXCL5, CXCL6, CXCL8, CCL3, and CCL4. Conclusions: TRL is a potent negative prognostic factor in ICI-treated mUC. IL13RA2 emerges as a central driver linking leukocytosis to immune resistance by promoting inflammatory signaling, cytokine production, and myeloid recruitment. These findings identify IL13RA2 as a key mediator of TRL and a potential therapeutic target to overcome myeloid-driven ICI resistance in urothelial carcinoma. Citation Format: Harvey Yu-Li Su, Shih-Yu Huang, Chung-Wen Kuo, Chang-Ting Lin, Yi-Hua Chen, Ming-Chun Kuo, Hsuan-Ying Huang, Chih-Yen Chien. Multi-omics dissection of tumor-related leukocytosis in urothelial carcinoma identifies IL13RA2 as a key driver [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 5570.

This report describes a rare primary subglottic adenoid cystic carcinoma harboring a SMARCA4 loss-of-function mutation, a molecular driver not previously reported in this anatomical site. Despite presenting with locally advanced pT4a disease and early pulmonary metastasis, the patient achieved 5-year local control following total laryngectomy and adjuvant radiotherapy. The findings suggest that molecular profiling can identify non-MYB-driven subtypes and guide individualized management for atypical laryngeal malignancies.

Background To evaluate the prognostic significance of preoperative monocyte-to-albumin ratio (MAR) in patients with oral squamous cell carcinoma (OSCC). Methods This study enrolled 408 patients diagnosed with OSCC who underwent primary radical surgery between 2011 and 2017. Preoperative MAR was calculated as the ratio of absolute monocyte count (∗106/L) to serum albumin level (g/L), measured within one week before surgery, while univariate and multivariate survival analyses were conducted using Kaplan-Meier and Cox proportional hazards regression models. A nomogram incorporating various clinicopathological factors was developed to predict overall survival (OS) on an individualized basis. Results The 5-year rates for OS, cancer-specific survival (CSS), and relapse-free survival (RFS) were 81.6%, 88.9% and 82.7% respectively. The optimal cut-off value for MAR was determined to be 8.31. Patients with high MAR values had significantly lower 5-year OS (75.1% vs. 91.3%, p < 0.001), CSS (83.5% vs. 96.9%, p < 0.001) and RFS (77.0% vs. 91.1%, p < 0.001) compared to those with low MAR values. In Cox regression analysis, high MAR independently predicted worse outcomes for OS (HR: 1.91, 95% CI: 1.1–3.317, p = 0.022), CSS (HR: 3.43, 95% CI: 1.329–8.852, p = 0.011) and RFS (HR: 2.282, 95% CI: 1.245–4.183, p = 0.008). The MAR-based nomogram demonstrated good predictive accuracy for OS (concordance index: 0.751). Conclusion The preoperative MAR is a valuable prognostic marker in OSCC. Integration of MAR into nomogram-based risk assessment models could enhance the stratification of patients at higher risk of adverse oncological outcomes, thereby informing treatment decisions.

OBJECTIVE:To evaluate the prognostic significance of the margin-to-depth of invasion ratio (MDR) in pathological nodal negative (pN0) oral tongue squamous cell carcinoma (OTSCC). STUDY DESIGN:Case series with chart review. SETTING:Tertiary academic medical center. METHODS:Patients with pN0 OTSCC who underwent primary radical surgery and high-quality neck dissection (lymph node yield ≥18) without positive margin (<1 mm) between 2007 and 2017 were retrospectively analyzed. Cox proportional hazards models were utilized to identify factors associated with survival outcomes, including cancer-specific survival (CSS) and relapse-free survival (RFS). RESULTS:A total of 296 patients were enrolled in this study. The optimal cutoff value for MDR was determined to be 0.29. For those with high MDR, the 5-year CSS and RFS were significantly superior compared to those with low MDR (CSS: 92.8% vs 71.5%, P < .001; RFS: 89.3% vs 67.9%, P < .001). In the multivariate Cox model, a low MDR remained an independent negative prognosticator of CSS (HR: 2.846, 95% CI: 1.229-6.593, P = .015) and RFS (HR: 2.661, 95% CI: 1.126-6.288, P = .026). In radical surgery without adjuvant therapy subgroup patients, patients with a low MDR had significantly poorer CSS and RFS than patients with a high MDR (both P < .001). CONCLUSION:Among pN0 OTSCC patients with negative surgical margins (≥1 mm), survival outcomes differed significantly between the high and low MDR groups. Patients with a low MDR had a markedly higher risk of recurrence and poorer cancer-specific survival, suggesting that adjuvant treatment may be warranted for this subgroup.

BACKGROUND:This study evaluated the prognostic significance of achieving a textbook outcome (TO) in patients with surgically treated p16-negative oropharyngeal squamous cell carcinoma (OPSCC). METHODS:A total of 194 patients with p16-negative OPSCC who underwent radical surgery and neck dissection between 2007 and 2017 were included. Cox proportional hazards models were applied to identify factors associated with overall survival (OS) and disease-free survival (DFS). RESULTS:Among the cohort, 100 patients (51.5%) achieved a TO. Patients in the TO group demonstrated significantly better 5-year OS rates (76.5% vs. 52.3%, p < 0.001) and 5-year DFS rates (72.2% vs. 47.2%, p = 0.001) compared with those in the non-TO group. Multivariate analysis revealed that failure to achieve a TO be an independent negative predictor of OS (hazard ratio [HR] 1.824, 95% confidence interval [CI] 1.126-2.957, p = 0.015) and DFS (HR 1.662, 95% CI 1.044-2.645, p = 0.032). CONCLUSION:Failure to achieve a TO is associated with poorer long-term outcomes in surgically treated patients with p16-negative OPSCC. Therefore, TO may serve as a practical, composite proxy for evaluating and improving surgical quality in this population.

PURPOSE:To prospectively compare dose to swallowing-related organs-at-risk (OARs) and swallowing outcomes between proton (intensity-modulated proton therapy, IMPT) and photon (volumetric modulated arc therapy, VMAT) radiotherapy (RT), and to characterize dose-response relationships associated with dysphagia in nasopharyngeal carcinoma (NPC). METHODS:In total, 104 patients with NPC received definitive chemoradiotherapy (IMPT, n = 58; VMAT, n = 46). Mean OAR doses were compared. Swallowing-related quality of life (QoL) was assessed using the MD Anderson Dysphagia Inventory (MDADI) at baseline (T1), end of RT (T2), and one-year post-RT (T3). Logistic regression evaluated predictors of ≥ 10-point MDADI decline or > 10% weight loss; linear mixed-effects and multivariable linear models assessed OAR dose-response relationships. RESULTS:IMPT delivered lower mean doses to the oral cavity, middle pharyngeal constrictor muscle (M-PCM), and inferior pharyngeal constrictor muscle (all P < 0.01), with similar sparing of salivary glands and the superior pharyngeal constrictor muscle (S-PCM). IMPT resulted in less weight loss at T2 (3.4% vs 5.5%; P = 0.019) and lower odds of > 10% loss (OR 0.10; 95% CI, 0.01-0.80). MDADI Composite and domain scores did not differ by RT modality. Higher baseline MDADI predicted better T2 and T3 scores, and older age reduced the risk of ≥ 10-point decline at T2. Higher S-PCM dose was consistently associated with worse MDADI at T2 and T3, while M-PCM dose was associated with acute declines at T2. CONCLUSIONS:IMPT improves sparing of several swallowing-related OARs and reduces early weight loss, but swallowing-related QoL is associated with dose at pharyngeal constrictor muscles, particularly the S-PCM.

PURPOSETo quantify risk reduction in oral precancer and cancer with time since cessation of betel quid chewing, smoking, and alcohol drinking.PATIENTS AND METHODSWe conducted a multicenter case-control study in Taiwan of patients with invasive oral cancer (n = 768), visual/clinical oral precancer (n = 1,998), and hospital-based controls (n = 717, recalibrated/reweighted to represent the Taiwan general population). Weighted logistic regression was used to evaluate associations of duration, intensity, and time since cessation of betel quid chewing without tobacco, smoking, and alcohol drinking (modeled using splines and categorically) with oral precancer and cancer risk versus controls.RESULTSRisk of oral precancer increased with increased duration and intensity of chewing and smoking, and increased intensity of alcohol drinking. Risk of oral cancer increased with increased duration and intensity of chewing, increased duration of smoking, and increased intensity of alcohol drinking. For oral precancer, when compared with current users (defined as current users and individuals who quit for ≤2 years before study recruitment), odds ratios (ORs) for 10-year cessation of behaviors were as follows: chewing = 0.63 (bootstrap 95% CI, 0.43 to 0.83); smoking = 0.30 (95% CI, 0.18 to 0.41); and alcohol = 0.60 (95% CI, 0.34 to 0.86). Likewise, risk of oral cancer decreased with increasing time since cessation; ORs for 10-year cessation were as follows: chewing = 0.74 (95% CI, 0.44 to 1.05); smoking = 0.45 (95% CI, 0.22 to 0.67); and alcohol drinking = 0.55 (95% CI, 0.20 to 0.90). Adjusted population-attributable fractions show that cessation of chewing for ≥10 years would prevent an estimated 23.2% of oral precancers and 24.2% of oral cancers; cessation of smoking for ≥10 years would prevent 51.5% of oral precancers and 44.6% of cancers in Taiwan.CONCLUSIONOur findings support the potential for primary prevention of oral precancer and cancer through cessation of betel quid chewing (without tobacco), smoking, and alcohol drinking among current users.

Head and neck squamous cell carcinoma (HNSC) ranks among the most common malignancies globally, with ALDH2 mutations linked to elevated disease risk. This study delineates the tumor-suppressive functions of ALDH2, examining its enzymatic activity, post-translational modifications, potentially transcriptional regulation, and protein–protein interactions. In an oral squamous cell carcinoma (OSCC) cohort, high ALDH2 immunostaining independently correlated with improved clinical outcomes. Functional assays across four HNSC-derived cell lines revealed that ALDH2 inhibits anchorage-independent growth, migration, invasion, and endothelial tube formation effects mediated by suppression of the HRAS–AKT–NFκB signaling axis. Mutations at E504 and phosphorylation-deficient variants at T261 and S488 impaired ALDH2 enzymatic function and abolished its tumor-suppressive capacity by reactivating oncogenic signaling. Mechanistically, ALDH2-mediated inhibition of AKT1 reduced NR4A1 phosphorylation, thereby enhancing TGM2 transcription and translation and promoting apoptosis. Notably, ALDH2 directly interacts with ALDH6A1, and this association, independent of catalytic activity, synergistically amplifies anti-tumor signaling. Collectively, these findings identify ALDH2 as a key tumor suppressor in HNSC, including OSCC, and highlight the therapeutic potential of activating ALDH2, NR4A1, and TGM2. Moreover, stabilization of the ALDH2–ALDH6A1 complex may offer a viable strategy for disease prevention and treatment, even in the context of frequent ALDH2 mutations.

BACKGROUND:The survival benefit of adjuvant radiotherapy/chemoradiotherapy (RT/CRT) in pT4aN0M0 gingival squamous cell carcinoma (GSCC) with adequate surgical margins remains unclear due to limited subsite-specific evidence. In this retrospective cohort study, we evaluated the associations of adjuvant RT/CRT with survival outcomes and identified prognostic factors in this clinically relevant subgroup. METHODS:We used the Taiwan Cancer Registry (2011-2021) to identify patients with first primary T4aN0M0 GSCC and clear resection margins (≥5 mm). Patients treated with surgery alone were compared with those receiving adjuvant RT/CRT. Cohorts were balanced via 1:1 propensity score matching (PSM) on age, sex, tumor depth, tumor differentiation, and comorbidity burden. Disease-specific survival (DSS) and overall survival (OS) were assessed using Kaplan-Meier estimates and Cox proportional hazards regression models. RESULTS:Among the 530 study patients, 75.3% (n = 399) received adjuvant RT/CRT. In the unmatched cohort, 5-year DSS and OS rates were similar for surgery alone vs. adjuvant RT/CRT (80% vs. 79%, p = 0.27; 67% vs. 72%, p = 0.99, respectively). After PSM (125 pairs), the 5-year DSS and OS rates were comparable, being 81% vs. 83% (p = 0.61) and 68% vs. 78% (p = 0.46), respectively. Multivariable analysis demonstrated that increasing age, moderate/poor differentiation, depth > 14 mm, and Charlson comorbidity index > 1 were independent prognosticators for DSS or OS, whereas adjuvant RT/CRT was not independently associated with better outcomes. CONCLUSION:For patients with T4aN0M0 GSCC and clear surgical margins, adjuvant RT/CRT did not confer a survival advantage over surgery alone, suggesting limited benefit of routine adjuvant therapy in this setting. Prospective studies are needed to validate this finding and to refine evidence-based risk stratification models.

BACKGROUND:Systemic inflammation and nutritional status are key determinants of prognosis in oral squamous cell carcinoma (OSCC). The neutrophil-to-lymphocyte ratio and serum albumin in peripheral blood are established markers reflecting host immune response and nutritional condition, respectively. The lymphocyte-albumin-neutrophil ratio (LANR), a composite index integrating these parameters, has emerged as a potential prognostic biomarker in several cancers; however, its role in OSCC remains unclear. This study aimed to evaluate the prognostic value of preoperative LANR in OSCC. METHODS:We retrospectively analyzed OSCC patients who underwent radical surgery between 2007 and 2017, randomly dividing them into training and validation cohorts by a 2:1 ratio. LANR was calculated by multiplying the absolute lymphocyte count by the serum albumin level and dividing by the absolute neutrophil count. The primary outcomes were 5-year overall survival (OS) and disease-free survival (DFS). Survival-associated factors were identified through univariate and multivariate Cox regression analyses, serving as the foundation for constructing a predictive nomogram model. RESULTS:This study included a total of 660 patients, with 440 assigned to the training cohort and 220 to the validation cohort. A LANR cutoff value of 18.7 effectively stratified patients into distinct prognostic groups, demonstrating significant differences in survival outcomes. A low LANR independently predicted worse OS (hazard ratio [HR]: 1.54, 95% CI: 1.04-2.28; p = 0.03) and DFS (HR: 1.54, 95% CI: 1.08-2.19; p = 0.017). The LANR-based nomogram demonstrated strong predictive performance, with concordance indices of 0.75 and 0.71 in the training and validation cohorts, respectively. CONCLUSION:Preoperative LANR serves as an independent prognostic marker in patients with OSCC. Incorporating LANR into risk models may enhance patient stratification and guide treatment strategies.

Objectives. This study aimed to evaluate the prognostic value of the margin-to-depth of invasion ratio (MDR) in patients with locally advanced oral squamous cell carcinoma (LAOSCC) who underwent curative surgery followed by adjuvant concurrent chemoradiotherapy (CCRT).Methods. We analyzed 422 consecutive patients with LAOSCC treated at a single institution between 2007 and 2017. MDR was defined as the ratio of the closest surgical margin (mm) to the tumor depth of invasion (mm). Survival outcomes, including overall survival (OS), cancer-specific survival (CSS), and relapse-free survival (RFS), were assessed. The optimal MDR cutoff was determined by X-tile analysis and validated using repeated k-fold cross-validation.Results. The optimal MDR cutoff for predicting survival was 0.35. Patients with MDR ≥0.35 (high MDR, n=205) demonstrated significantly better 5-year OS (66.1% vs. 47.6%, P<0.001), CSS (77.5% vs. 57.4%, P<0.001), and RFS (71.5% vs. 53.8%, P=0.001) than those with MDR <0.35 (low MDR, n=217). In multivariate analysis, low MDR remained an independent adverse prognostic factor for OS (hazard ratio [HR], 1.612; P=0.005), CSS (HR, 2.028; P=0.001) and RFS (HR, 1.501; P=0.033). Among patients with adequate margins (≥5 mm), MDR retained significant prognostic value (OS, P=0.008; CSS, P=0.001; RFS, P=0.015). Cross-validation confirmed the robustness of the MDR threshold value of 0.35 across all survival endpoints.Conclusion. MDR is an independent prognostic marker in LAOSCC treated with surgery and adjuvant CCRT. A cutoff of 0.35 effectively stratifies survival risk, even among patients with adequate surgical margins. Incorporating MDR into postoperative assessment could refine risk stratification and guide individualized follow-up and adjuvant treatment planning.

OBJECTIVES:To assess the prognosis of young patients (≤40 years old) with head and neck squamous cell carcinoma (HNSCC), focusing on the preoperative Systemic Inflammation Response Index (SIRI). METHODS:Between January 2007 and February 2017, 175 young patients with HNSCC (≤40 years old) who underwent radical surgery were retrospectively enrolled in this study. The patients were randomly divided into a training cohort (N = 131) and a validation cohort (N = 44). The SIRI is defined as the absolute neutrophil count (×10⁹/L) multiplied by the absolute monocyte count (×10⁹/L), divided by the absolute lymphocyte count (×10⁹/L) in peripheral blood, all measured within one week prior to radical surgery. Univariate and multivariate Cox regression analyses were conducted to identify variables associated with survival outcomes, which were then used to construct and evaluate a predictive nomogram. RESULTS:In both the training and validation cohorts, patients were classified into low- and high-SIRI groups based on a cutoff value of 0.87, which was determined by receiver operating characteristic analysis. This SIRI cutoff effectively stratified patients into two distinct prognostic groups with significant survival differences. Multivariable Cox analysis identified the presence of lymphovascular invasion and the high preoperative SIRI as significant independent prognostic factors associated with poorer cancer-specific survival (CSS) in young patients with HNSCC. Using these variables, a predictive model for 5 year CSS was constructed and visualized as a nomogram. The model demonstrated strong predictive performance, with a C-index of 0.744 [95% CI (0.643-0.845)] in the training cohort and 0.839 [95% CI (0.740-0.938)] in the validation cohort. CONCLUSION:Data from preoperative SIRI assessment, coupled with the presence of pathological adverse features, serve as valuable references for risk stratification in young patients with HNSCC.

Head and neck squamous cell carcinoma (HNSCC) remains a prevalent and challenging cancer to treat due to its genetic heterogeneity. Cisplatin resistance is one of important causes in treatment failure of locally advanced HNSCC. ONC201, a selective dopamine receptor D2 antagonist and mitochondrial ClpP agonist, has emerged as a potential antitumor agent in various malignancies. This study explores the therapeutic potential of ONC201, alone and in combination with cisplatin, in both cisplatin-sensitive and -resistant HNSCC cells, with an emphasis on endoplasmic reticulum (ER) stress-mediated apoptosis. A cisplatin-resistant HNSCC subline (OC2-CR1) was developed via long-term drug exposure. The treatment effectiveness of ONC201 alone and cisplatin in combination on cell viability, DNA damage, reactive oxygen species (ROS) production, and stress response markers were evaluated. ONC201 exhibited potent cytotoxicity in both cisplatin-sensitive and -resistant HNSCC cells, retaining efficacy in OC2-CR1 cells. Combined treatment with ONC201 and cisplatin demonstrated synergistic inhibition of proliferation and migration, with enhanced induction of apoptosis. Mechanistically, ONC201 induced ER stress-mediated cell death via ATF4/CHOP signaling in cisplatin-sensitive cells, while ATF3/CHOP predominated in resistant cells. In vivo, combination therapy significantly suppressed tumor growth in xenograft models, including cisplatin-resistant tumors, without inducing toxicity. Immunohistochemical analysis confirmed activation of CHOP in tumor tissues. Furthermore, clinical correlation revealed that low CHOP expression in OSCC patients was associated with increased recurrence risk and inferior recurrence-free survival significantly. This study provides compelling evidence that ONC201 enhances cisplatin efficacy through distinct, stress-mediated apoptotic pathways in HNSCC. The ability of ONC201 to overcome cisplatin resistance and its synergistic antitumor effects highlight its promise as a candidate for combination therapy. These findings support the translational potential of targeting the ATF3/ATF4/CHOP axis to improve outcomes in patients with cisplatin resistant HNSCC.

Background:This study aimed to evaluate the survival predictability of preoperative systemic inflammation response index (SIRI), calculated as the absolute neutrophil count multiplied by the absolute monocyte count and divided by the absolute lymphocyte count, in patients with metachronous secondary primary head and neck squamous cell carcinoma (mspHNSCC) who had undergone prior radiotherapy for first primary HNSCC (fpHNSCC). Methods:A total of 101 consecutive patients who underwent upfront surgery for mspHNSCC at a single institute between 2007 and 2016 were retrospectively reviewed between December 2023 and November 2024 and included in the analysis. The baseline leukocyte counts for the fpHNSCC and mspHNSCC were collected. Cox proportional hazards models were constructed using age and variables significant in univariate analysis to assess the impact of SIRI on overall survival (OS) and cancer-specific survival (CSS). Additionally, a SIRI-based nomogram was developed and validated. Results:Statistically significant declines in baseline leukocyte counts were observed in mspHNSCC compared to fpHNSCC (p < 0.001). Among the inflammatory markers, the preoperative SIRI was the most predictive of survival outcomes for mspHNSCC. Higher SIRI values were significantly associated with poorer outcomes in both OS and CSS. The optimal SIRI cutoff for survival prediction was 1.383, as determined by receiver operating characteristic curve analysis with Youden's index; patients with SIRI ≥ 1.383 had significantly lower 5-year OS (32.9% vs 60.1%, p = 0.001) and CSS (64.7% vs 83.9%, p = 0.003). Multivariate analysis revealed lymphovascular invasion, extranodal extension, and high SIRI as independent adverse risk factors for CSS. The SIRI-based nomogram accurately predicted CSS, with a concordance index of 0.773. Conclusion:Data from preoperative SIRI assessment, coupled with the presence of pathological adverse features, serve as valuable references for risk stratification in patients with previously irradiated mspHNSCC.

Oral tongue squamous cell carcinoma (OTSCC) is an aggressive malignancy and the most common subsite of head and neck cancer among Taiwanese males. This study aimed to evaluate the prognostic significance of depth of invasion (DOI) in patients with node-negative OTSCC treated with radical surgery alone. We retrospectively analyzed 243 patients with node-negative OTSCC who had undergone radical surgery with adequate margins between 2005 and 2017. Each millimeter increase in DOI was significantly associated with a higher hazard of all-cause mortality (ACM) (hazard ratio [HR], 1.07; 95% confidence interval [CI], 1.03-1.111; p < 0.001), cancer-specific mortality (CSM) (HR, 1.087; 95% CI, 1.04-1.136; p < 0.001) and local recurrence (LR) (HR, 1.081; 95% CI, 1.021-1.145; p = 0.008), but not regional recurrence (RR) (HR, 1.042; 95% CI, 0.986-1.102; p = 0.144). In multivariate analysis, DOI remained an independent predictor of ACM, CSM, and LR. A DOI-based nomogram demonstrated improved predictive performance, with a concordance index of 0.700 for overall survival. In conclusion, DOI represents a crucial prognostic factor for ACM, CSM, and LR in patients with node-negative OTSCC treated with surgery alone, highlighting its potential clinical utility for early risk stratification and guidance in decision-making regarding adjuvant therapy or intensified surveillance.

Oral cavity squamous cell carcinoma (OSCC), a leading subtype of head and neck cancer, exhibits high global incidence and mortality rates. Despite advancements in surgery and radiochemotherapy, approximately one-third of patients experience relapse. To improve current targeted and immunotherapy strategies for recurrent OSCC, we conducted multi-omics analyses on pretreatment OSCC samples (cohorts 1 and 2, n=137) and identified A3A and EGFR, both at the RNA and protein levels, as inversely expressed markers for patient stratification and response prediction. Survival analysis demonstrated that elevated A3A or PD-L1 expression levels correlated to improved responses to anti-PD-1 therapy in patients (cohort 3a, n=50, IHC). In contrast, high RRAS expression (cohort 4, n=252, qRT-PCR) was significantly associated with OSCC recurrence. Cell-based experiments revealed that RRAS was involved in radiotherapy and cisplatin resistance through the EGFR/RRAS/AKT/ERK signaling pathway. In OSCC patient-derived xenograft (PDX) mouse models, treatments with cisplatin and cetuximab (anti-EGFR) effectively reduced tumor size in EGFR-high-derived (#34) but not A3A-high-derived (#22) PDX tumors. Our study demonstrated that A3A-high tumors were immune-hot and responsive to anti-PD-1 therapy, whereas EGFR-high tumors exhibited chr.7p11.2 gains and DNA repair alterations. Additionally, RRAS-high tumors were associated with OSCC recurrence via AKT and ERK phosphorylation and demonstrate improved clinical outcomes with cetuximab therapy (cohort 3b, n=49, IHC). This study emphasizes the significance of A3A and EGFR expression levels in OSCC patient stratification and precision therapy, suggesting the use of anti-PD-1 or anti-EGFR treatments, respectively based on these biomarkers. Furthermore, RRAS emerges as a novel prognostic marker for local recurrence.