BACKGROUND:Non-small cell lung cancer (NSCLC) is a particularly aggressive subtype of lung cancer characterized by early metastasis and poor prognosis. Lung adenocarcinoma (LUAD) represents the most prevalent histological subtype within NSCLC. The development of brain metastases in LUAD is frequently associated with a severely unfavorable outcome. While extrachromosomal circular DNA (eccDNA) has been implicated in various tumors, its specific role in brain metastasis related to LUAD remains largely unexplored. METHODS:EccDNA associated with brain metastasis in LUAD profiles was collected through sequencing 20 cerebrospinal fluid (CSF) samples from both control subjects and LUAD patients with brain metastases, focusing on those with Epidermal Growth Factor Receptor (EGFR) mutations, rare mutations, or no mutations at all. The genomic characteristics of the eccDNAs were examined across groups utilizing a circular map. Differentially expressed eccDNAs were subjected to analysis and functional annotation by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG), employing eccDNA-related genes. RESULTS:The distribution of eccDNAs within the genome is intricately associated with gene density. We compared the genomic features of eccDNAs between the subtypes of control and brain metastatic LUAD. Differentially expressed eccDNAs in CSF were identified, and subsequent GO and Kyoto Encyclopedia of Genes and Genomes (KEGG) analyses demonstrated their functional relevance in each group. CONCLUSIONS:Our findings offer initial insights into the features of CSF-enriched eccDNAs across LUAD subtypes with varying pathogenic mechanisms, emphasizing their potential as diagnostic and prognostic indicators for LUAD.

Combining immune checkpoint inhibitors (ICI) with chemotherapy has been established as the standard first-line (1 L) treatment for advanced non-small cell lung cancer (NSCLC). However, the optimal second-line (2 L) treatment following 1 L chemo-immunotherapy remained controversial. Patients with advanced NSCLC who progressed following 1 L chemo-immunotherapy and continued to receive ICI as 2 L therapy were enrolled in the study. These patients were divided into two groups according to the therapeutic modality: chemo-immunotherapy plus anti-angiogenesis therapy group (CIA group) and chemo-immunotherapy group (CI group). Baseline characteristics were balanced using inverse probability of treatment weighting (IPTW) to minimize selection bias before comparative analyses. The efficacy of immunotherapy rechallenge combination therapy was evaluated based on overall survival (OS) and progression-free survival (PFS). Cox proportional hazards regression analyses were applied to determine predictive factors independently associated with survival outcomes. A total of 101 patients were enrolled in this study, 45 patients were enrolled in CIA group, and those who didn’t receive anti-angiogenesis drugs were defined as CI group (n = 56). Overall, Immunotherapy rechallenge reached a median OS of 19.47 months and median PFS of 7.10 months. The CIA group showed significantly longer PFS than the CI group (11.49 vs. 6.06 months, p = 0.03). Similar results in PFS were observed in the study cohorts balanced with IPTW (11.49 vs. 6.89, p = 0.01). Multivariate analyses revealed that prior immunotherapy response and smoking status could be independent prognostic factors for PFS. Immunotherapy rechallenge could bring survival benefits following progression under chemo-immunotherapy, especially those who responded to prior immunotherapy. Additionally, the use of anti-angiogenesis drugs could significantly improve the clinical response of immunotherapy rechallenge.

Radiation-induced lung injury (RILI) arises from the unavoidable exposure of normal lung tissue during radiotherapy for thoracic malignancies. It remains a dose-limiting complication in thoracic radiotherapy, critically impacting treatment efficacy and long-term patient survival. Despite its clinical significance, the dynamic molecular reprogramming underlying RILI progression remains poorly characterized. Utilizing a C57BL/6J thoracic irradiation mouse model validated by histopathological analysis, we implemented an integrative tri-omics approach (RNA-seq, LC-MS/MS proteomics, and UHPLC-QTOF metabolomics) to systematically map the temporal evolution of RILI at critical phases: acute inflammation, transitional, and fibrotic. The three periods correspond to 4 weeks, 8 weeks, and 16 weeks after the mice received 16 Gy of electron beam irradiation to the thoracic region. Combined analysis using complete-linkage hierarchical clustering revealed co-expression modules, and Ces2e was identified as a pivotal node exhibiting sustained upregulation at both transcriptional and translational levels during early pathogenesis. Functional enrichment revealed time-dependent activation of xenobiotic metabolism and extracellular matrix (ECM)-receptor interaction pathways, with Ces2e overexpression correlating with macrophage polarization and lipid peroxidation accumulation. This temporal multi-omics atlas not only deciphers stage-specific molecular signatures of RILI but also nominates Ces2e as a candidate early-stage molecular marker.

Resistance to 5-fluorouracil-based chemotherapies presents a major clinical challenge in colorectal cancer (CRC) treatment, leading to elevated recurrence rates. Recent findings indicate that enhanced metabolic plasticity in cancer cells plays a crucial role in drug resistance development, though the exact mechanisms remain uncertain. In this study, we demonstrate that the lipophagy-triggered fatty acid oxidation (FAO) is involved in maintaining the adenosine triphosphate (ATP) levels upon drug administration. Using mass spectrometric imaging, we identified arachidonic acid (AA) as a critical lipid mediator of intracellular selective autophagy and energy balance in resistant cancer cells. The high level of arachidonate metabolism was associated with poor response to chemotherapy in clinical practice. Mechanistically, accumulated AA induced phosphorylation of PLINs via the LOXs/PKCζ pathway and stimulated the production of free fatty acid (FFA) driven by lipophagy. Importantly, we further certified that inhibition of AA-associated lipophagy specifically sensitize drug-resistant colorectal cancer cells to chemotherapy. This study offers insights into the mechanism of acquired chemoresistance, suggesting that targeting AA-associated lipophagy could be a potential therapeutic strategy to overcome drug resistance in colorectal cancer.

Developing effective therapeutic strategies for head and neck squamous cell carcinoma (HNSCC) remains a considerable clinical challenge. Cetuximab, a first-line targeted therapy for HNSCC, exhibits limited efficacy. The aim of this study was to explore the potential of α-1,3-mannosyltransferase (ALG3) inhibition in augmenting the therapeutic efficacy of cetuximab. We first analyzed the Cancer Genome Atlas (TCGA) data and found that ALG3 was significantly overexpressed in HNSCC tissues, correlating with worse pathological features and lower overall and disease-specific survival. Functional studies using ALG3-knockdown cells and a subcutaneous tumor model demonstrated that ALG3 inhibition markedly suppressed HNSCC proliferation both in vitro and in vivo. Furthermore, combining ALG3 inhibition with cetuximab elicited potent anti-cancer effects in vitro and in vivo. Mechanistic investigations via quantitative polymerase chain reaction, western blotting, and transmission electron microscopy revealed that ALG3 knockdown induced endoplasmic reticulum (ER) stress in HNSCC cells through the Bip/IRE1α axis. Finally, blocking N‑linked glycosylation synergistically enhanced cetuximab-mediated growth inhibition of HNSCC cells. In conclusion, ALG3 is a promising target to enhance the therapeutic efficacy of cetuximab in HNSCC.

Background: The optimal approach to treatment intensification for epidermal growth factor receptor (EGFR)-mutant lung adenocarcinoma with brain metastases remains a topic of debate, especially in the context of high-risk molecular features such as TP53 co-mutation. The extent to which baseline clinical risk influences treatment efficacy is yet to be determined. Objectives: We aim to systematically evaluate the prognostic significance of EGFR/TP53 co-mutations in treatment-naïve patients with lung adenocarcinoma and newly diagnosed brain metastases. Design: This study enrolled 218 treatment-naïve patients with EGFR-mutant lung adenocarcinoma and brain metastases who received first-line third-generation EGFR-tyrosine kinase inhibitors (TKIs). Treatment effects were evaluated using interaction models stratified by Lung-molGPA scores. Stratification: Group A (Lung-molGPA 1–2) versus Group B (Lung-molGPA 2.5–4). Methods: Within each group, the influence of TP53 co-mutations on survival was analyzed. Additionally, among patients with TP53 co-mutations, the effects of cranial radiotherapy (CRT) and treatment with either third-generation EGFR-TKIs monotherapy or third-generation EGFR-TKIs combined with chemotherapy on overall survival (OS) were further investigated, taking into account the Lung-molGPA scores. Results: The Lung-molGPA significantly influenced the prognostic impact of TP53 mutations and the survival benefits of treatment intensification strategies (log-rank test p = 0.044, hazard ratio (HR) = 2.1832, 95% confidence interval (CI): 1.023–4.661). In patients with low Lung-molGPA scores, intensification with CRT did not correlate with a survival advantage (32.2 vs 28.0 m, log-rank test p = 0.5088, HR = 1.417, 95% CI: 0.4768–4.211). Conversely, in patients with high Lung-molGPA scores, both CRT and systemic intensification were associated with improved outcomes (44.0 vs 26.3 m, log-rank test p = 0.0054, HR = 0.3199, 95% CI: 0.1585–0.6456; 20.1 vs 32.3 m, log-rank test p = 0.0074, HR = 2.547, 95% CI: 1.113–5.831). Conclusion: Baseline clinical risk, as delineated by the Lung-molGPA, serves as a crucial determinant of therapeutic benefit in cases of EGFR-mutant lung adenocarcinoma with cerebral metastases. Implementing risk-adapted treatment intensification strategies could potentially prevent overtreatment in patients classified as low-risk, while simultaneously optimizing clinical outcomes in high-risk cohorts.

ObjectiveTo externally evaluate and update the QUANTEC and Appelt NTCP models for radiation pneumonitis (RP) in lung cancer patients treated with contemporary IMRT and multimodal therapy, and to preliminarily validate a simplified local model in an independent cohort.MethodsWe retrospectively analyzed 580 lung cancer patients treated with thoracic IMRT between 2018 and 2023 as the development cohort. The QUANTEC and Appelt models were evaluated and locally updated using a closed testing procedure to determine the least extensive revision required. Clinical and DVH variables were standardized, and smoking status and pulmonary comorbidity were recoded according to published definitions. A final simplified local model (Model D) was developed using BIC-guided multivariable logistic regression with regularization. Performance was assessed by AUC, Brier score, calibration-in-the-large (CITL), calibration slope, Hosmer–Lemeshow test, and decision curve analysis. External validation of Model D was performed in 100 patients from an independent center using fixed coefficients.ResultsBoth the QUANTEC and Appelt models showed substantial calibration bias in the local cohort, with systematic underestimation of RP risk. Updating improved calibration as expected, with little change in discrimination. Model D, incorporating age, stage, smoking status, tumor location, pulmonary comorbidity, NLR, SII, V30, and MLD, showed the best apparent overall performance in the development cohort (AUC 0.708, Brier 0.215, CITL = 0, slope = 1, Hosmer–Lemeshow P = 0.599). In the external cohort, discrimination and prediction error were similar (AUC 0.718, 95% CI 0.576–0.831; Brier 0.207), although absolute RP risk was overestimated (CITL = −1.043, slope = 1.133, Hosmer–Lemeshow P < 0.001).ConclusionsThe original QUANTEC and Appelt models underestimated RP risk in this contemporary IMRT cohort. Updating improved calibration, whereas discrimination changed little. Model D showed better apparent overall performance and preserved ranking ability in an independent external cohort. Calibration drift across centers suggests that simple recalibration may improve absolute risk estimation in new settings.Clinical trial registrationhttps://www.chictr.org.cn/hvshowproject.html?id=276191&v=1.1, identifier ChiCTR2500102055.

Lung cancer remains the leading cause of cancer-related mortality worldwide, and immune checkpoint blockade is often limited by an immunosuppressive tumor microenvironment (TME). To address these challenges, we developed a novel multifunctional nanoplatform, based on a mesoporous Fe3O4 core and silica shell, co-loading pemetrexed and an anti-PD-L1 antibody (PD-L1&Pem@msNPs). The construct enables checkpoint-targeted delivery, alternating magnetic field (AMF)-triggered hyperthermia and controlled drug release, enabling a singlesystem chemo-immuno-magnetothermal regimen. We comprehensively investigated the physicochemical properties, magnetothermal performance, loading/conjugation, and release behavior, and validated cellular uptake and cytotoxicity in vitro. In subcutaneous and orthotopic lung tumor models, PD-L1&Pem@msNPs achieved superior tumor suppression and extended survival compared with control formulations. Mechanistically, transcriptomic profiling together with immunophenotyping demonstrated marked TME remodeling, with increased intratumoral T-cell representation accompanied by coordinated rewiring of macrophages status. Notably, longitudinal flow cytometry revealed a shift in T-cell states from an exhausted-intermediate (Tex-int) toward a progenitor-like (Tex-prog) phenotype, consistent with restoration of T-cell functionality under treatment. Collectively, PD-L1&Pem@msNPs provides an externally activatable, modular platform to remodel the TME and improve the therapeutic impact of PD-L1 blockade in lung cancer.

Developing effective therapeutic strategies for colorectal cancer (CRC) presents a significant clinical challenge. Although immune checkpoint blockade therapies, such as the CD47 blockade, have shown promise, their efficacy is often limited by suboptimal responses and associated toxicities. This study explores the potential of the gut microbial metabolite cyclic diadenosine monophosphate (c-di-AMP) to enhance the therapeutic efficacy of CD47 antibodies in treating CRC while mitigating its adverse effects. A subcutaneous tumor model was established to assess the therapeutic efficacy of c-di-AMP in combination with CD47 blockade for CRC treatment. Growth inhibition assays were performed to evaluate the direct effects of c-di-AMP on CRC cells. Cell removal assays, flow cytometry, and immunofluorescence were employed to investigate the impact of c-di-AMP on macrophage-mediated phagocytosis. Macrophage polarization phenotypes were characterized by flow cytometry and immunohistochemistry. Transcriptome sequencing was also conducted to elucidate the molecular mechanisms underlying the c-di-AMP-induced enhancement of macrophage phagocytic activity. Our results revealed that c-di-AMP, in combination with CD47 blockade, exerted potent anticancer effects in vivo. Mechanistically, c-di-AMP enhanced macrophage-mediated phagocytosis triggered by CD47 blockade by promoting M1 polarization and activating the NF-κB signaling pathway. Furthermore, c-di-AMP upregulated the expression of CD11b/CD18, thereby strengthening intercellular adhesion between macrophages and CRC cells, which further facilitated phagocytosis. These findings indicate that c-di-AMP may enhance the efficacy of CD47 blockade in CRC by promoting macrophage phagocytosis through NF-κB activation.

​Background: Neoadjuvant immunochemotherapy achieves higher pathological complete response (pCR) rates than chemotherapy alone in limited-stage Small Cell Lung Cancer (LS-SCLC). However, whether pCR translates into a survival benefit remains unclear, and the underlying biological mechanisms have yet to be investigated.Patients and Methods: This is a multicenter retrospective cohort study. 85 patients with stage I-III LS-SCLC who underwent radical resection following neoadjuvant immunochemotherapy (NIC, n=28) or neoadjuvant chemotherapy (NC, n=57) were included. The primary objectives were to compare pCR rates between treatment groups and to assess the prognostic impact of pCR stratified by treatment group, using event-free survival (EFS) and overall survival (OS) as clinical endpoint. Seven-plex multiplex immunofluorescence (mIF) was performed on nine surgical specimens selected via purposive sampling. Public transcriptomic datasets (GSE281525, GSE60052) served as independent validation.Results: The pCR rate in the NIC group was significantly higher than the NC group (35.7% vs 14.0%, P=0.027). With the median follow-up of 20.8 months in the NIC group, pCR patients had longer EFS than non-pCR patients (not reached vs 20.3 months, P=0.007). No such difference was seen in the NC group (19.8 vs 18.0 months, P=0.945). Spatial immune profiling revealed that NIC pCR tumors were uniformly immune-hot (2/2) with high CD8+ T cell infiltration and low PD-1 exhaustion. Nearest-neighbor distance analysis revealed closer macrophage-CD8 T cell co-localization in NIC pCR compared with NC pCR specimens. Transcriptomic analysis of independent cohorts confirmed that CD8 activation and composite antitumor immune scores were significantly associated with favorable survival.Conclusions: pCR after neoadjuvant immunochemotherapy, but not chemotherapy, translates to survival benefit in LS-SCLC. Spatial immune profiling reveals distinct immune-mediated versus drug-mediated mechanisms underlying SCLC tumor clearance. These findings highlight the need to consider treatment type when interpreting pCR in SCLC and provide rationale for integrating spatial immune biomarkers in prospective neoadjuvant trials.

Background Non-small cell lung cancer brain metastases (NSCLC-BM) confer a poor prognosis, with only a minority of patients responding to immune checkpoint inhibitors (ICIs). Tertiary lymphoid structures (TLSs) have been linked to prognosis and response to immunotherapy across various tumours. Given the unique immune environment of brain metastases, TLSs within NSCLC-BM might exhibit distinct characteristics. Nevertheless, the function and formation mechanisms of these intracranial TLSs remain inadequately understood. Methods We retrospectively analysed 120 resected NSCLC-BM samples. Heterogeneity of TLS characteristics and functional states in NSCLC-BM was assessed using H&E and multiplex immunohistochemistry (mIHC). Associations with OS and iPFS were evaluated, and a nomogram incorporating TLS score was developed. Potential mechanisms underlying TLS formation were explored using public scRNA-seq datasets and mIHC, and were functionally validated in vivo in the LC-BrM mouse model. Findings TLSs were primarily intratumoral and immature. Elevated intratumoral TLS scores independently predicted prolonged OS and iPFS, and predicted benefit from postoperative ICIs. TLS-rich tumours exhibited increased CD8+ T cells, FOXP3-CD4+ T cells, and B cells. Mechanistically, TCF1+CD4+ T cells and microglia co-localised within TLSs and might facilitate their formation via LTβ/LTβR signalling. Meanwhile, stromal CXCL12 was further identified as a crucial chemoattractant for these cells and correlated with favourable outcomes of ICI therapy. Functionally, the combination of rmCXCL12 and anti-PD-1 promotes TLS formation and intracranial tumour control in an LTβR-dependent manner. Interpretation These findings highlight the significance of TLSs as prognostic biomarkers and potential therapeutic targets in patients with NSCLC-BM. Funding This work was supported by the Natural Science Foundation of China (82573031); the CQMU Program for Youth Innovation in Future Medicine (W0172); the Postdoctoral Fellowship Program of the China Postdoctoral Science Foundation (GZC20233356).

Radiotherapy (RT) has emerged as an effective synergistic therapy to enhance PD-1 inhibitor efficacy in Lung cancer brain metastases (LCBM). Nonetheless, not all patients benefit from this radioimmunotherapy strategy. Further research is imperative to determine the optimal RT fractionation regimen for enhancing PD-1 inhibitor efficacy in LCBM. An intracranial LCBM model was established by intracranial injection of LLC-Luc-BrM cells. Mice received PD-1 inhibitors combined with RT delivered at the same biologically effective dose (BED) using three RT fractionation regimens: conventionally fractionated radiotherapy (CFRT, 2 Gy × 8f), hypofractionated radiotherapy (HFRT, 3 Gy × 5f), and single-fraction high-dose radiotherapy (SF-HDRT, 10 Gy × 1f). Intracranial tumor growth was monitored by IVIS imaging, and survival was assessed using Kaplan–Meier analysis. T cell infiltration and function were evaluated using mIHC and ELISA. Cell depletion experiments were performed to clarify the essential cell populations involved in this synergistic efficacy. To investigate endothelial adhesion molecule expression and tertiary lymphoid structure (TLS) formation-related programs, RNA sequencing, IHC, and mIHC were employed. Among the three regimens, HFRT combined with PD-1 inhibitors most effectively inhibited tumor growth and prolonged survival. This combination significantly enhanced infiltration of CD4 + CD3 + and CD8 + CD3 + T cells and increased secretion of GZMB, IFN-γ, and TNF-α. T cell depletion experiments confirmed that this antitumor efficacy depended on both CD8 + T cells and CD4 + T cells. Mechanistically, HFRT upregulated ICAM-1 and VCAM-1 on CD31 + endothelial cells and promoted perivascular accumulation of CD3 + T cells, consistent with enhanced vascular permissiveness. HFRT also induced TLS-like aggregates and enriched TLS-associated gene signatures, with a nonclassical phenotype. HFRT was identified as an optimal RT fractionation regimen for enhancing PD-1 inhibitor efficacy in LCBM. The synergistic efficacy requires both CD4 + and CD8 + T cells and is associated with vascular remodeling and nonclassical TLS formation induced by HFRT.

Recent studies have applied machine-learning radiomics to predict EGFR mutations or PD-L1 expression, yet little is known about PD-L1 prediction specifically in EGFR-mutant lung adenocarcinoma, despite evidence linking high PD-L1 expression to TKI resistance. To address this gap, this study integrates CT radiomics features with clinical data to develop and validate interpretable models for predicting PD-L1 expression in advanced EGFR-mutant LUAD and identifying key predictive determinants. In this single-center study, we retrospectively collected the pretreatment CT images, including non-contrast enhanced CT (NECT) and contrast enhanced CT (CECT), as well as clinical information from a cohort consisting of 165 EGFR-Mutant LUAD patients consecutively staged at IIIB-IVB between January 2020 and May 2022. After feature extraction and selection, six machine learning algorithms (Logistic regression, Linear discriminate analysis, Supporter vector machine, Gradient boosting, Random forest and adaptive boosting) were employed to construct the models. Model performances were assessed using 5-fold cross-validation and then visualized using coefficient matrices and SHAP (SHapley Additive exPlanations) to interpret the models’ predictions. A total of 30 radiomics features and 4 clinical features were selected. Models using radiomics features alone achieved the highest area under the curve (AUC) value of 0.91, and the combination of radiomics and clinical features models showed similar predictive performance with an AUC value of 0.90. However, the clinical features models exhibited insufficient predictive ability and poor generalizability (AUC = 0.64). According to the coefficient matrices, the top 5 features associated with PD-L1 ≥ 1

BackgroundEmerging evidence has demonstrated that lung cancer patients with brain oligometastases (oligo-BMs) could benefit from local radical therapies. Despite rapid advancements in the treatment of oligometastatic disease, the molecular determinants governing the oligometastatic phenotype in the central nervous system remain elusive.MethodsWe performed 1021-panel sequencing, transcriptome profiling, DNA methylation mapping, and multiplex immunohistochemistry on 20 paired primary lung adenocarcinoma and oligo-BMs specimens to delineate their evolutionary dynamics and microenvironmental characteristics.ResultsA pronounced intertumor heterogeneity was observed between primary tumors (PTs) and oligo-BMs, while intratumoral heterogeneity was conserved across lesions. Subclonal analysis and phylogenetic reconstruction demonstrated that oligo-BMs exhibited predominant polyclonal seeding patterns and parallel progression trajectories. Moreover, oligo-BMs displayed a more immunosuppressed microenvironment compared to PTs, characterized by attenuated immunogenic cell death signatures, downregulation of immune-activated pathways, and diminished infiltration of activated immune cells (including B cells, NK cells, and Th1 cells). The methylation levels at functional genomic regions were highly concordant between PTs and oligo-BMs. Notably, we identified NLGN1 as a potential regulator of oligo-BM, where the hypermethylation at its genebody regions was mechanistically associated with transcriptional upregulation. Clinical validation revealed that NLGN1 was specifically overexpressed in BMs compared to PTs and extracranial metastases, correlating with advanced pathological stages and poor prognosis. In vivo studies further confirmed NLGN1 promotes BM in mice.ConclusionsOur findings provide novel insights into the evolutionary trajectory, immune landscape and methylation pattern of oligo-BMs, potentially paving the way for the development of innovative therapeutic strategies for lung cancer patients with oligo-BMs.

Abstract Background: Dysregulation of MET gene could serve as oncogenic driver in NSCLC. The efficacy of MET inhibitor monotherapy was limited in EGFR wild-type NSCLC patients(pts) with MET alterations. Pre-clinical study demonstrated that MET inhibitor combined with immunotherapy may synergistically enhance anti-tumor activity. SOUND study aims to explore the efficacy and safety of savolitinib plus durvalumab in EGFR wild-type advanced NSCLC pts with MET alterations. Methods: Pts who did not receive immunotherapy previously were enrolled into MET Ex14m cohort (Cohort 1) or MET Overexpression/AMP cohort (Cohort 2) according to MET alteration status determined by NGS, FISH or immunohistochemistry and received savolitinib (600mg [body weight≥50kg] / 400mg [body weight < 50kg], once daily) in combination with durvalumab (1,500 mg once every four weeks). The primary endpoint was progression-free survival (PFS). Secondary endpoints included objective response rate (ORR), 12-month overall survival (OS) rate, and safety. Here, we report the interim analysis results. Results: At data cutoff (January 17, 2025), forty-seven pts were enrolled and received at least one dose of study treatment, with 24 in Cohort 1(C1) and 23 in Cohort 2(C2), respectively. For C1, 83.3% pts had adenocarcinoma, 16.7% had brain metastases, 37.5% had PD-L1 ≥ 50% and 87.5% were treated in first line. For C2, 56.5% had adenocarcinoma, 34.8% had brain metastases, 56.5% had PD-L1 ≥ 50% and 87.0% were treated in first line. Median follow-up was 15.4 months(m) in C1 and 13.2 m in C2. The median PFS was not reached (maturity of 33.3%) in C1 and 5.5 m (95% CI, 3.2-9.2, maturity of 78.3%) months in C2. The confirmed ORR per investigator in C1 and C2 were 45.8% (95% CI, 25.6-67.2%) and 52.2% (95% CI, 30.6-73.2%), respectively. 12-month OS rate for C1 and C2 were 82.0% and 60.1%, respectively. Any grade treatment-related adverse events (TRAEs) were observed in 44 (93.6%) pts. The most common TRAEs were peripheral oedema (38.3%), anemia (38.3%), hypoalbuminemia (34%), aspartate aminotransferase increased (34%) and alanine aminotransferase (ALT) increased (31.9%). Grade 3 and above TRAEs were reported in 14 (58.3%) pts in C1 and 12 (52.2%) pts in C2. The most common grade 3 and above TRAE were hepatic function abnormal (14.9%), drug-induced liver injury (12.8%) and ALT (10.6%) increased. Conclusions: Savolitinib plus durvalumab showed promising clinical anti-tumor activity in pts with advanced EGFR wild-type NSCLC with MET alterations, especially in pts with MET ex14m. Close and frequent(i.e. weekly) monitoring of liver function should be conducted in pts receiving this combination. Prospective studies with larger sample sizes are needed to further evaluate the efficacy and safety of this combination in EGFR wild-type NSCLC pts with MET alterations. Clinical trial information: NCT05374603 Citation Format: Yong-Feng Yu, Xiao-Ying Huang, Qian Chu, An-Wen Liu, Li Zhuang, Xiao-Rong Dong, Hong-Cheng Wu, Jian-Ying Zhou, Shun-Dong Cang, Yan Wang, Yi Hu, Zhen-Zhou Yang, Meng-di Wu, Xiao-yuan Wang, Shun Lu. Savolitinib combined with Durvalumab in EGFR wild-type advanced NSCLC patients with MET alterations (SOUND): A multicenter, open-label, Phase II trial [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(8_Suppl):Abstract nr CT247.

BACKGROUND:Neoadjuvant immunochemotherapy achieves higher pathological complete response (pCR) rates than chemotherapy alone in limited-stage small cell lung cancer (LS-SCLC), but whether pCR carries similar prognostic information across treatment modalities is uncertain. PATIENTS AND METHODS:This multicenter retrospective cohort study included 85 patients with stage I-III LS-SCLC who underwent radical resection after neoadjuvant immunochemotherapy (NIC, n = 28) or chemotherapy (NC, n = 57). pCR rates were compared between regimens, and treatment-stratified associations of pCR with event-free survival (EFS) and overall survival (OS) were assessed. Treatment-by-pCR interactions were examined using Firth-penalized Cox models. Seven-plex multiplex immunofluorescence (mIF) in nine purposively selected specimens and two public transcriptomic cohorts provided exploratory immune context. RESULTS:The pCR rate was 35.7% for NIC and 14.0% for NC (P = 0.022). At a median NIC follow-up of 21.1 months, pCR was associated with longer EFS (NR vs 20.3 months, P = 0.007). No corresponding association was detected in the NC group (18.0 vs 19.8 months, P = 0.892). The treatment-by-pCR interaction was significant (Firth-penalized Cox, P = 0.004), indicating that the association between pCR and EFS differed between treatment groups. In the descriptive mIF analysis, both selected NIC pCR specimens showed an immune-hot phenotype, characterized by the highest CD8 + T cell infiltration among the specimens examined and lower PD-1 exhaustion than the NIC non-pCR specimens. Higher antitumor immune scores were associated with longer OS in GSE60052. CONCLUSIONS:In this selected surgical cohort, pCR was more frequent after NIC than NC and showed different EFS associations across treatment groups in exploratory analyses. Both examined NIC pCR specimens were immune-hot, but these mIF observations are hypothesis-generating and cannot establish a mechanism. These findings require prospective validation.

Nuclear factor of activated T-cells, cytoplasmic 4 (NFATc4), a transcription factor of the NFAT family, has been reported to participate in the tumorigenesis and progression of several cancers. However, the function and regulation of NFATc4 in lung adenocarcinoma (LUAD) remain poorly understood. Here, we report for the first time that NFATc4 is significantly overexpressed in LUAD tissues, and high NFATc4 expression correlates with lymphatic metastasis, advanced tumor stage, and poor prognosis in patients. Subsequent functional studies revealed that NFATc4 depletion inhibits LUAD cell viability, proliferation, and tumor growth by inducing cell cycle arrest in the G2/M phase and apoptosis. A mechanistic study shows that NFATc4 knockdown leads to significant enrichment of cellular process-related pathways and differentially expressed genes, especially downregulated genes Cyclin B1 (CCNB1) and cyclin-dependent kinase 1 (CDK1). NFATc4 directly binds to the CCNB1 promoter to regulate the CCNB1/CDK1 pathway, resulting in cell cycle arrest and inhibition of cell proliferation. This study identifies NFATc4/CCNB1/CDK1 as a novel regulatory pathway involved in LUAD development and provides a potential prognostic biomarker and molecular therapeutic target for LUAD.

INTRODUCTION:Cerebrospinal fluid (CSF) metabolites are implicated in various neurological disorders, but their relationship with primary brain malignancies remains unclear. This study aims to use a two-sample Mendelian randomization (MR) approach to evaluate the role of CSF metabolites in the development of primary brain malignancies. METHODS:Data on 338 human CSF metabolites were obtained from a genome-wide association study (GWAS) involving 291 participants. The information on primary brain malignancies was derived from a large-scale GWAS summary, which included 816 cases and 314,193 controls of European ancestry. The inverse-variance weighted model was used as the primary analysis method, supplemented by sensitivity analyses such as heterogeneity testing, horizontal pleiotropy testing, and leave-one-out analysis. Linkage disequilibrium score regression was performed for genetic correlation analysis. RESULTS:This study identified ten CSF metabolites whose associations with primary brain malignancies are consistent with a potential causal role (0.0001 < P < 0.05), including one unidentified metabolite. Of these, five metabolites were positively correlated with the risk of disease, while four showed a negative correlation. Notably, after FDR correction, the causal association between elevated levels of 5-methylthioadenosine (MTA) and increased risk of primary brain malignancies remained (P = 0.038), and there was no evidence of reverse causality between the two, suggesting that MTA may be an important risk factor for primary brain malignancies. CONCLUSIONS:The findings provide evidence consistent with potential causal associations between ten CSF metabolites and primary brain malignancies, helping us to further understand the pathogenesis of primary brain malignancies and provide clues for the prediction of the disease.

RATIONALE:Human epidermal growth factor receptor 2 (HER2)-positive cholangiocarcinoma is a rare disease with a low incidence and high degree of malignancy. Trastuzumab deruxtecan (T-DXd) has been approved for the treatment of HER2-positive breast and gastric cancer. However, it is still in the initial exploration period for HER2-positive cholangiocarcinoma. PATIENT CONCERNS:A 57-year-old Han Chinese male patient with recurrent metastatic cholangiocarcinoma who was tested for HER2 expression in surgical specimens in the absence of reliable drug therapy. DIAGNOSES:Postoperative pathological examination confirmed a diagnosis of moderately to poorly differentiated adenocarcinoma of the common bile duct. INTERVENTIONS:Our case revealed HER2 amplification and effectively received T-DXd as backline therapy. OUTCOMES:After approximately 4 months without disease progression, the patient experienced an increase in plasma tumor markers; however, he regained disease control after receiving T-DXd in combination with lenvatinib with a favorable physical status and quality of life. LESSONS:Reexamination of tumor tissue samples to identify target mutations is necessary for backline treatment of cholangiocarcinoma. T-DXd is effective in the treatment of HER2 amplification cholangiocarcinoma and is relatively well tolerated after multiple lines of therapy. The combination of multi-target tyrosinase inhibitors is a possible strategy for overcoming resistance in the future.