Objective: Neoadjuvant chemotherapy regimens have shown encouraging efficacy characterized by high objective response rate (ORR), pathologic complete response (pCR) rate, and major pathologic response (MPR) rate, alongside acceptable safety. This single-center retrospective study aimed to evaluate the safety and efficacy of neoadjuvant pembrolizumab plus chemotherapy in patients with locally advanced resectable oral and oropharyngeal squamous cell carcinomas (LA-OSCC/OPSCC). Materials and methods: A total of 50 patients were included. The patients received 2-4 cycles of neoadjuvant therapy with pembrolizumab, albumin-bound paclitaxel and cisplatin before surgery, followed by adjuvant radiotherapy or immunotherapy. Results: The median follow-up time was 31.7 months (95%CI, 29.4-34.0). The ORR was 85.4%, and the MPR rate was 65.8%. The 1-year event-free survival (EFS) rate was 88.8% (95%CI, 79.8%-98.8%). Patients with moderate programmed cell death ligand 1 (PD-L1) expression (combined positive score (CPS) 1 to <10) achieved the highest MPR rate (71.4%), underscoring the potential predictive value of PD-L1 expression. Treatment-related adverse events (TRAEs), most commonly alopecia, anemia, neutropenia, and nausea, were manageable. No treatment-related deaths occurred. Conclusion: This retrospective analysis indicates that neoadjuvant pembrolizumab combined with chemotherapy is a promising strategy for patients with LA-OSCC/OPSCC. Future prospective studies with larger cohorts and longer follow-up are warranted to confirm these findings.

Hepatocellular carcinoma (HCC) is a global health challenge, with limited treatment options for advanced-stage patients. Although recent approvals of targeted therapies and immune checkpoint inhibitors (ICIs) have expanded the therapeutic landscape, their clinical benefits are often constrained by modest response rates and acquired resistance. Despite ongoing efforts to identify new therapeutic targets, only few inhibitors have progressed to clinical trials, benefiting a small subset of patients. These limitations underscore the need for more effective therapeutic strategies. We integrated data from DepMap and TCGA to identify HCC-specific vulnerabilities. A kinome-wide CRISPR screen validated essential targets. Pharmacological inhibition (AZD1152) and genetic knockout of Aurora B (AURKB) were employed for functional validation. Senescence was assessed via SA-β-gal staining, transcriptomics, and SASP analysis. Immune interactions were evaluated using co-culture assays with T cells and flow cytometry for MHC I and immune markers. In vivo efficacy was tested in immunocompetent murine HCC models. Aurora B was identified as a top candidate essential for HCC survival, overexpressed in tumors, and correlated with poor prognosis. A kinome-wide CRISPR screen further confirmed its essential role in HCC cell survival. Pharmacologic inhibition or genetic knockout of Aurora B markedly suppressed proliferation of HCC cells and induced robust cellular senescence, characterized by cell-cycle arrest, DNA damage, and SASP. These senescent cells exhibited heightened susceptibility to T-cell-mediated cytotoxicity, associated with increased immunoproteasome activity and upregulated MHC I expression. Co-culture assays revealed that senescent tumor cells promoted interferon-γ production and enhanced cytotoxicity in CD8⁺ T cells as well as PD-1 expression. In both in vitro and in vivo models, combining Aurora B inhibition with anti-PD-1 therapy markedly suppressed tumor growth. This study identifies Aurora B as a therapeutically actionable vulnerability in HCC, whose inhibition induces senescence and enhances tumor immunogenicity. The combination of Aurora B inhibitor with anti-PD-1 therapy yields synergistic anti-tumor effects, supporting its potential as a promising strategy to improve immunotherapy outcomes in HCC.

Human papillomavirus (HPV)-negative head and neck squamous cell carcinoma (HNSCC) is characterised by hyperactivation of the cyclin-dependent kinase 4/6 (CDK4/6) pathway. As immunotherapy has become the first-line treatment for HNSCC, resistance to anti-programmed death-1 (PD-1) agents has emerged as a pivotal challenge. This prospective, single arm, phase II study (NCT05721443) evaluated the efficacy and safety of dalpiciclib, a CDK4/6 inhibitor, combined with cetuximab in patients with anti-PD-1-resistant, HPV-negative recurrent and/or metastatic HNSCC. Patients diagnosed with p16-negative R/M HNSCC resistant to first-line anti-PD-1 therapy without prior cetuximab treatment were enroled. Patients received oral dalpiciclib (150 mg daily on days 1-21 of each 28-day cycle) and intravenous cetuximab (400 mg/m2 on day 1 of cycle 1, followed by 250 mg/m2 weekly in each cycle). The primary endpoint was objective response rate (ORR), secondary endpoints were overall survival, progression-free survival, duration of response, and safety. Between March 2023 and November 2024, a total of 28 patients were enroled. The ORR was 67.9

INTRODUCTION:Oral mucosal melanoma (OMM) is a rare subtype of melanoma but exhibits highly invasive biological behavior. Programmed cell death protein 1 (PD-1) monotherapy showed lower response in OMM than other subtypes of melanoma. MATERIALS AND METHODS:We retrospectively analyzed the efficacy and safety of pembrolizumab with anlotinib in patients with advanced OMM between August 2018 and September 2024. The primary endpoint was objective response rate (ORR); the secondary endpoints included disease control rate (DCR), duration of response (DOR), progression-free survival (PFS), overall survival (OS), and treatment-related adverse events (TRAEs). RESULTS:Forty-one patients were enrolled in our study. Seventeen patients (41.4%) achieved an objective response. The median PFS was 6.4 months (95% confidence interval [CI], 4.5-8.3 months), and the median OS was 10.0 months (95% CI, 8.3-11.7 months). Thirty-three patients (80.5%) experienced at least one TRAE. The most common TRAEs were hypertension (29.3%), hand-foot syndrome (19.5%), and anemia (19.5%). Grade 3 or higher TRAEs occurred in two patients (4.8%), and no grade 5 TRAEs were observed. CONCLUSIONS:Our study suggested that pembrolizumab with anlotinib in patients with advanced OMM showed potential efficacy and manageable adverse effects. Further studies are needed to confirm the efficacy of this combination strategy.

Hepatocellular carcinoma (HCC) remains a therapeutic challenge due to the limited efficacy of current systemic therapies. To identify potential therapeutic approaches, computational analysis of HCC datasets and drug screening were integrated, leading to the repurposing of hematological malignancy drug selinexor (an XPO1 inhibitor) for HCC treatment. Functional studies revealed that XPO1 inhibition triggers oxidative stress and cell cycle arrest in HCC cells through nuclear sequestration of NCOR1, disrupting redox homeostasis through FOXK1-dependent transcriptional activation of genes associated with reactive oxygen species (ROS). A genome-wide CRISPR-Cas9 screen further identified the KEAP1-NRF2 axis as a key determinant of sensitivity to XPO1 inhibition. Furthermore, high-throughput compound screening demonstrated that disulfiram, a clinically used aldehyde dehydrogenase inhibitor, synergizes with XPO1 inhibitor through exacerbation of ROS accumulation. Collectively, these findings demonstrate the therapeutic repurposing of selinexor for HCC while uncovering its mechanism of action, establishing a predictive biomarker, and proposing an immediately translatable combination therapy.

Mucosal melanoma (MM), an aggressive melanoma subtype arising in mucosal tissues, displays resistance to therapies effective in cutaneous melanoma. To understand how mucosal microenvironment contributes to treatment nonresponsiveness, we performed integrative analysis of single-cell and bulk messenger RNA sequencing data derived from oral mucosa-originated melanoma and revealed that mucosa-specific inflammation induces enrichment of low-pigmented neural crest-like cancer cell, mediated by COX2+ macrophages and their secretome. Maintenance of this inflammation-induced neural crest-like state in cancer cells depends on HER2 and HER3 activation. Inhibition of HER2/3 by pan-HER inhibitors blocks cell state plasticity and overcomes chemoresistance in primary MM cell lines and patient-derived xenograft (PDX) models. These findings provide insights into how the tissue of origin determines cancer aggressiveness, highlight the role of mucosal inflammation in driving melanoma stemness and chemoresistance, and advance the identification of effective treatment options currently lacking for patients with MM.

Objective·To compare the immune microenvironment of mucosal malignant melanoma(MM)and cutaneous malignant melanoma(CM)using single-cell transcriptome analysis system,and to elucidate the key regulatory mechanism of CD8+T cell depletion in MM.Methods·A total of 36 531 cells from three treatment-naïve MM surgical specimens and three CM samples were subjected to single-cell RNA sequencing.Data pre-processing included batch correction,strict quality control(based on mitochondrial gene ratio and gene number screening),unsupervised clustering,and cell-type annotation(using established marker genes).Differential gene analysis(Wilcoxon test),gene set enrichment analysis[based on Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)databases],cell-cell interaction network analysis,and prognostic association analysis using.The Cancer Genome Atlas(TCGA)were used to comprehensively analyse the heterogeneity of MM and CM tumour immune microenvironment.Results·Compared with CM,the expression of cytotoxicity-related genes(GZMB,IFNG,etc.)in CD8+T cells in MM was reduced(P＜0.001),while the depletion markers(PDCD1,LAG3,etc.)were significantly up-regulated(P＜0.001).The proportion of depleted CD8+T cell subsets increased by nearly 6 times(CM:3.38%vs MM:19.26%,P＜0.010),and the terminal depletion score was higher.In addition,the level of neutrophil infiltration in the MM microenvironment was significantly higher than that in CM(CM:1.6%vs MM:34.1%,P=0.009),and single-cell analysis further divided it into five subgroups.Among them,the CD10+neutrophil subset(accounting for 23.52%of MM neutrophils)highly expressed pro-inflammatory molecules(S100A8/A9)and immunosuppressive molecules(MME/CD10,CD55),and the characteristic scores of myeloid-derived suppressor cells(MDSCs)in this subset were significantly higher than those of other subgroups.TCGA and MM cohort analyses further confirmed that the highly expressed genes of this subset were independently associated with poor prognosis(P=0.049),suggesting that this subset was involved in tumour immunosuppressive effects.Mechanistically,CD10+neutrophils interacted with depleted CD8+T cells through the SELPLG-SELL ligand-receptor pair,driving T-cell inactivation.Conclusion·Single-cell transcriptome sequencing was performed on MM clinical samples,and the single-cell immunomes of MM and CM were systematically compared,suggesting that CD10+neutrophils in MM induce CD8+T-cell terminal depletion through the SELPLG-SELL pathway.The molecular mechanism of MM immunotherapy tolerance was elucidated,providing a new strategy for targeting neutrophil-T cell interaction.

Cryoablation therapy for tumors has a long history of clinical application. Its anti-tumor mechanisms and histopathological changes have been well established, with extensive clinical practice demonstrating its safety and efficacy, theoretically making it an ideal modality for tumor treatment. Historically constrained by limitations in cryogenic media and freezing equipment, its therapeutic effectiveness and clinical adoption were significantly restricted. The emergence of new-generation cryoablation systems represented by Argon-Helium cryosurgical systems has achieved substantial advancements in refrigeration efficiency, ablation range precision, and temperature monitoring accuracy, thereby greatly promoting the widespread adoption of tumor cryoablation technology. This consensus systematically summarizes the mechanisms of cryoablation technology, indications for cryotherapy in head and neck mucosal melanoma, standardized clinical treatment protocols, management of adverse reactions, and related principles. It aims to provide authoritative references for standardizing cryoablation therapy in the treatment of head and neck mucosal melanoma.

While combination BRAF/MEK inhibition has improved survival in BRAFV600 mutant melanoma, targeted therapies for BRAFWT melanoma remain limited. Microphthalmia transcription factor (MITF), a lineage-specific transcription factor that regulates melanocyte proliferation and melanin synthesis, represents a promising melanoma-specific drug target. In this study, we evaluated TT-012, a recently identified MITF dimerization specific inhibitor, and surprisingly found that most BRAFWT melanoma lines were resistant to TT-012 due to low MITF transcriptional activity and reduced dependency on MITF for proliferation. High-throughput drug screen identified tivozanib, an FDA-approved drug targeting VEGFR and other receptor tyrosine kinases (RTKs), which sensitized cells to TT-012. Mechanistically, tivozanib induced cell state transition from MITFlow to MITFhigh state via VEGFR2 inhibition followed by NF-κB pathway activation, restoring MITF transcriptional activity and growth dependency. The combination of tivozanib and TT-012 synergistically inhibited melanoma growth both in vitro and in vivo, underscoring its potential as a novel therapeutic strategy for BRAFWT melanoma.

Ferroptosis, a unique form of iron-dependent cell death triggered by lipid peroxidation accumulation, holds great promise for cancer therapy. Despite the crucial role of GPX4 in regulating ferroptosis, our understanding of GPX4 protein regulation remains limited. Through FACS-based genome-wide CRISPR screening, we identified MALT1 as a regulator of GPX4 protein. Inhibition of MALT1 expression enhances GPX4 ubiquitination-mediated degradation by up-regulating the E3 ubiquitin ligase RC3H1. Using both rescue assays and functional genetic screening, we demonstrate that pharmacologically targeting MALT1 triggers ferroptosis in liver cancer cells. Moreover, we show that targeting MALT1 synergizes with sorafenib or regorafenib to induce ferroptosis across multiple cancer types. These findings elucidate the modulatory effects of the MALT1-RC3H1 axis on GPX4 stability, revealing a molecular mechanism that could be exploited to induce ferroptosis for cancer therapy.

Mucosal melanoma (MM) is a rare and aggressive form of melanoma with a poorer prognosis compared to other subtypes. Recent large-scale next-generation sequencing studies, including our own research, have demonstrated that the molecular characteristics and potential oncogenic drivers of MM differ significantly from those of cutaneous melanoma. The emergence of selective CDK4/6 inhibitors, already approved for use in breast cancer and undergoing phase III clinical trials for other solid tumors, represents a promising development in the treatment of MM. Recent studies have shown that CDK4/6 inhibitors not only induce cell cycle arrest but also play a crucial role in facilitating the interaction between tumor cells and the host immune system. Moreover, our findings indicate that dysregulation of cell cycle progression due to cyclin‐dependent kinase 4 (CDK4) amplification is a significant genetic characteristic in a substantial portion of MM cases. Targeting CDK4 in specific MM patients shows promise for precision cancer therapy, utilizing molecularly characterized MM patient-derived xenograft (PDX) models and clinical trials. This paper provides an overview of existing literature on CDK4/6 dysregulation in MM, as well as preclinical and clinical investigations on CDK4/6 inhibitors and potential combination therapies for MM treatment.

Congenital melanocytic nevi(CMNs)are skin lesions char-acterized by benign melanocytic proliferations and present at birth or shortly thereafter.Large and giant CMNs,with a projected adult size＞20 cm and 40 cm in diameter respectively,are more likely to develop into malignant melanoma.In most cases,melanoma arising from congen-ital melanocytic nevus(CMN)is particularly aggressive.

Tumor-initiating cells (TICs) possess the ability to evade anti-tumor immunity, potentially explaining many failures of cancer immunotherapy. Here, we identify CD49f as a prominent marker for discerning TICs in hepatocellular carcinoma (HCC), outperforming other commonly used TIC markers. CD49f-high TICs specifically recruit tumor-promoting neutrophils via the CXCL2-CXCR2 axis and create an immunosuppressive milieu in the tumor microenvironment (TME). Reciprocally, the neutrophils reprogram nearby tumor cells toward a TIC phenotype via secreting CCL4. These cells can evade CD8+ T cell-mediated killing through CCL4/STAT3-induced and CD49f-stabilized CD155 expression. Notably, while aberrant CD155 expression contributes to immune suppression, it also represents a TIC-specific vulnerability. We demonstrate that either CD155 deletion or antibody blockade significantly enhances sensitivity to anti-PD-1 therapy in preclinical HCC models. Our findings reveal a new mechanism of tumor immune evasion and provide a rationale for combining CD155 blockade with anti-PD-1/PD-L1 therapy in HCC.

BackgroundThe KEYNOTE-048 and KEYNOTE-040 study have demonstrated the efficacy of pembrolizumab in recurrent or metastatic squamous cell carcinoma of the head and neck (R/M HNSCC), we conducted this real-world study to investigate the efficacy of pembrolizumab in patients with R/M HNSCC.MethodsThis is a single-center retrospective study conducted in the Shanghai Ninth People’s Hospital Affiliated to Shanghai Jiao Tong University School of Medicine (Shanghai, China). Between December 2020 and December 2022, a total of 77 patients with R/M HNSCC were included into analysis. The primary endpoint of the study was overall survival (OS), and the secondary endpoints were progression-free survival (PFS), overall response rate (ORR)and toxicity.Efficacy was assessed according to RECIST version 1.1.SPSS 27.0 and GraphPad Prism 8.0 software were utilized to perform the statistical analysis.ResultsBy the cut-off date (February 28, 2023), the median OS,PFS and ORR were 15.97 months,8.53 months and 48.9% in patients treated with the pembrolizumab regimen in the first line therapy. Among these patients, 17 patients received pembrolizumab with cetuximab,and 18 received pembrolizumab with chemotherapy.We observed no significant differences between two groups neither in median OS (13.9 vs 19.4 months, P=0.3582) nor PFS (unreached vs 8.233 months, P= 0.2807). In the ≥2nd line therapy (n=30), the median OS, PFS and ORR were 5.7 months, 2.58 months and 20% respectively. Combined positive score (CPS) was eligible from 54 patients. For first line therapy, the median OS and PFS were 14.6 and 8.53 months in patients with CPS ≥1, and median OS and PFS were 14.6 and 12.33 months in patients with CPS ≥20. The immune-related adverse events (irAEs) were occurred in the 31 patients (31/77, 40.26%), and the most common potential irAEs were hypothyroidism (25.97%), and pneumonitis (7.79%).ConclusionOur real-world results indicated that pembrolizumab regimen is a promising treatment in patients with R/M HNSCC

Although treatment with taxanes does not always lead to clinical benefit, all patients are at risk of their detrimental side effects such as peripheral neuropathy. Understanding the in vivo mode of action of taxanes can help design improved treatment regimens. Here, we demonstrate that in vivo, taxanes directly trigger T cells to selectively kill cancer cells in a non-canonical, T cell receptor-independent manner. Mechanistically, taxanes induce T cells to release cytotoxic extracellular vesicles, which lead to apoptosis specifically in tumor cells while leaving healthy epithelial cells intact. We exploit these findings to develop an effective ther-apeutic approach, based on transfer of T cells pre-treated with taxanes ex vivo, thereby avoiding toxicity of systemic treatment. Our study reveals a different in vivo mode of action of one of the most commonly used chemotherapies, and opens avenues to harness T cell-dependent anti-tumor effects of taxanes while avoid-ing systemic toxicity.

Ferroptosis is a unique form of intracellular iron-dependent cell death that differs from apoptosis, necrosis, and autophagy. GPX4, an antioxidant defense enzyme, plays a pivotal role as regulator of ferroptosis. Extensive researches suggest that targeting GPX4 holds promise for cancer therapy. However, the current GPX4 inhibitors face challenges due to unfavorable drug-like properties, which hinder their progress in clinical development. In this study, we identified a novel inhibitor called MI-2, demonstrating potent ferroptosis-inducing capacity. Mechanistically, MI-2 effectively inhibits the activity of GPX4 by direct interaction. Furthermore, MI-2 promotes the degradation of GPX4 through its well-established target, MALT1. In multiple cancer models, MI-2 has demonstrated synergistic effects when combined with sorafenib or regorafenib, resulting in enhanced ferroptosis induction. These findings highlight the dual modulatory effects of MI-2 on GPX4 activity and stability, offering a promising starting point for the development of drug-like GPX4 inhibitors with translational potential.### Competing Interest StatementThe authors have declared no competing interest.

The total protein concentration was measured in three regions of cerebrospinal fluid (CSF) in the cat. Values obtained were: cortical subarachnoid fluid 53.2 mg/100 ml; cisterna magna fluid 18.0 mg/100 ml; and lumbar subarachnoid fluid 36.3 mg/100 ml. The influx of protein into these regions of CSF was measured during steady-state perfusions of artificial CSF. Corrected protein influx was highest in cortical-subarachnoid perfusions (29.4 μg/min); intermediate in ventricular perfusions (7.6 μg/min), and lowest in spinal subarachnoid perfusions (4.8 μg/min). Protein permeability in the spinal subarachnoid space was approximately four times lower than in the intracranial compartments, indicating that the high concentration of protein in the lumbar fluid results from a slower rate of removal of protein from this region. In contrast, the high concentration of protein in the cortical subarachnoid fluid resulted from a high rate of influx of protein, presumably from the adjacent brain extracellular space.

3593 Background: Genomics-based precision medicine has greatly improved how patients with cancer are being treated with targeted agents, but clinical-grade genomic biomarkers for chemotherapies are currently lacking. The chemotherapeutic trifluridine/tipiracil (FTD/TPI) is approved for the treatment of late-stage metastatic colorectal cancer (mCRC). We aimed to find genomic biomarkers to improve patient selection for FTD/TPI treatment in mCRC. Methods: In a discovery cohort of FTD/TPI-treated mCRC patients (n = 37), genome-wide somatic variants were tested for association with treatment duration and overall survival (OS). In vitro drug testing on isogenic cell lines and patient-derived mCRC organoids, as well as a re-analysis of the double-blind, placebo-controlled, phase 3 RECOURSE trial (n = 800) were performed to support our findings. Results: In the discovery cohort, KRAS codon G12 (KRAS G12 ) mutation status was the only significant genomic determinant of poor outcome of FTD/TPI treatment, which could be replicated in vitro by drug testing on isogenic cell lines and PDOs. In these models, KRAS G12 mutations were associated with increased resistance to FTD-induced (geno)toxicity in vitro. KRAS G12 -based resistance was absent for the closely related chemotherapeutic 5-FU. In the RECOURSE study, KRAS G12 mutations were predictive biomarkers for reduced OS benefit of FTD/TPI vs placebo (unadjusted interaction P= 0.0017, adjusted interaction P= 0.017). For patients with KRAS G12 mutations, OS was not significantly prolonged with FTD/TPI vs placebo (n = 279; HR, 0.97; 95% CI, 0.73−1.20; P= 0.85). An exploratory analysis showed that the KRAS G13 mutant subgroup demonstrated clearly prolonged OS with FTD/TPI vs placebo (n = 60; unadjusted HR, 0.29; 95% CI, 0.15−0.55; P< 0.001; adjusted HR, 0.20; 95% CI, 0.092−0.45; P< 0.001), which was significantly more pronounced as compared to the KRAS G12 mutant and KRAS WT populations (adjusted interaction P< 0.001 and P= 0.036, respectively). Conclusions: Together, KRAS G12 mutations were associated with reduced OS benefit of FTD/TPI treatment, with potential implications for ̃28% of patients with metastatic colorectal cancer now considered for treatment with FTD/TPI. Furthermore, our data show that genomics-based precision medicine may be possible for a subset of chemotherapies.