Patients with non‑small cell lung cancer (NSCLC) harboring epidermal growth factor receptor (EGFR)‑activating mutations can be treated with EGFR‑tyrosine kinase inhibitors (TKIs). Although EGFR‑TKI‑targeted drugs bring survival promotion in patients with EGFR mutations, drug resistance is inevitable, so it is urgent to explore new treatments to overcome drug resistance. In addition, wild‑type EGFR lacks targeted drugs, and new targeted therapies need to be explored. Ferroptosis is a key research direction for overcoming drug resistance. However, the role and mechanism of regulating ferroptosis in different EGFR‑mutant NSCLC types remains unclear. In the present study, H1975 (EGFR T790M/L858R mutant), A549 (EGFR wild‑type) and H3255 (EGFR L858R mutant) NSCLC cell lines were used. The expression of ferroptosis markers in these cell lines was detected using western blotting and reverse transcription‑quantitative PCR. Cell viability was determined using the MTT assay and reactive oxygen species (ROS) levels were measured using flow cytometry. The results showed that, compared with EGFR wild‑type/sensitive mutant cells, EGFR‑resistant mutant cells were more sensitive to the ferroptosis inducer, erastin. Furthermore, the mammalian target of rapamycin (mTOR) inhibitor, everolimus (RAD001), induced cell death in all three cell lines in a dose‑dependent manner. The ferroptosis inhibitor, ferrostatin‑1, could reverse cell death in EGFR‑resistant mutant and EGFR wild‑type cells induced by RAD001, but could not reverse cell death in EGFR‑sensitive mutant cells. Compared with EGFR wild‑type/sensitive mutant cells, EGFR‑resistant mutant cells were more sensitive to RAD001 combined with erastin. In addition, a high‑dose of RAD001 reduced the expression levels of ferritin heavy‑chain polypeptide 1 (FTH1), glutathione peroxidase 4 (GPX4) and ferroportin and significantly increased ROS and malondialdehyde (MDA) levels in EGFR‑resistant mutant and EGFR wild‑type cells. In the present study, GPX4 inhibitor only or combined with RAD001 inhibited the AKT/mTOR pathway in EGFR‑resistant mutant cells. Therefore, the results of the present study suggested that inhibition of the mTOR pathway may downregulate the expression of ferroptosis‑related proteins in EGFR‑resistant and EGFR wild‑type NSCLC cells, increase the ROS and MDA levels and ultimately induce ferroptosis.

Somatic variants play a crucial role in the occurrence and progression of cancer. However, in the absence of matched normal controls, distinguishing between germline and somatic variants becomes challenging in tumor samples. The existing tumor-only genomic analysis methods either suffer from limited performance or insufficient interpretability due to an excess of features. Therefore, there is an urgent need for an alternative approach that can address these issues and have practical implications. Here, we presented OncoTOP, a computational method for genomic analysis without matched normal samples, which can accurately distinguish somatic mutations from germline variants. Reference sample analysis revealed a 0% false positive rate and 99.7% reproducibility for variant calling. Assessing 2864 tumor samples across 18 cancer types yielded a 99.8% overall positive percent agreement and a 99.9% positive predictive value. OncoTOP can also accurately detect clinically actionable variants and subclonal mutations associated with drug resistance. For the prediction of mutation origins, the positive percent agreement stood at 97.4% for predicting somatic mutations and 95.7% for germline mutations. High consistency of tumor mutational burden (TMB) was observed between the results generated by OncoTOP and tumor-normal paired analysis. In a cohort of 97 lung cancer patients treated with immunotherapy, TMB-high patients had prolonged PFS (P = .02), proving the reliability of our approach in estimating TMB to predict therapy response. Furthermore, microsatellite instability status showed a strong concordance (97%) with polymerase chain reaction results, and leukocyte antigens class I subtypes and homozygosity achieved an impressive concordance rate of 99.3% and 99.9% respectively, compared to its tumor-normal paired analysis. Thus, OncoTOP exhibited high reliability in variant calling, mutation origin prediction, and biomarker estimation. Its application will promise substantial advantages for clinical genomic testing.

Background: The necessity of platinum-doublet chemotherapy in first-line immunotherapy for non-squamous non-small cell lung cancer (nsqNSCLC) with programmed death-ligand 1 (PD-L1) expression on less than 50% of tumor cells remains poorly investigated. Biomarkers predicting this necessity can guide chemotherapy-free treatment to minimize unnecessary toxicity. Methods: Treated with immune checkpoint inhibitor monotherapy (ICI-mono), chemotherapy, or combination (ICI-chemo), 790 low PD-L1-expressing nsqNSCLCs (in-house: n=83; public: n=707) were analyzed for development and validation of the interaction score for additional chemotherapy (ISAC). Transcriptomic (public, n=11) and multiplex immunofluorescence data (in-house, n=100) were analyzed to evaluate the immune microenvironment. Results: ICI-chemo, compared with ICI-mono, tended to prolong progression-free survival (PFS; HR=0.72, p=0.004) and overall survival (OS; HR=0.77, p=0.071) as first-line therapy in low PD-L1-expressing nsqNSCLCs. The added value of chemotherapy was observed in the ISAC-low subgroup (PFS: HR=0.48, p<0.001; OS: HR=0.53, p=0.001) rather than the ISAC-high subgroup (PFS: HR=1.08, p=0.65; OS: HR=1.14, p=0.56). This predictive utility was independent of tumor mutational burden and PD-L1 expression, indicated by subgroup and multivariable analyses. A high ISAC was associated with adaptive immune resistance reflected by more proinflammatory (eg, CD8⁺ T cells and M1 macrophages) rather than anti-inflammatory tumor-infiltrating immune cells (eg, M2 macrophages) and high expression of immune checkpoints except for PD-L1 (eg, programmed cell death protein-1). Conclusion: A high ISAC was identified as a significant predictor for virtually no added value of platinum-doublet chemotherapy for first-line ICI treatment in low PD-L1-expressing nsqNSCLC. Our findings may help refine personalized therapeutic strategies for nsqNSCLC, thereby improving efficacy and reducing undue toxicity.

Background: For limited-stage small cell lung cancer (LS-SCLC), immunotherapy combined with chemotherapy as induction followed by subsequent radiotherapy is still under clinical investigation. Some patients who receive treatment have undergone radiotherapy following immunochemotherapy in real-world. However, the preliminary efficacy and safety remain unknown. In this study, we will explore a novel treatment approach for LS-SCLC. Methods: This study analyzed 32 patients with LS-SCLC treated at Jilin Cancer Hospital between January 2022 and June 2024. Eligible patients received Immune checkpoint inhibitors (ICIs) combined with chemotherapy, followed by sequential or concurrent radiotherapy and maintenance chemo-immunotherapy. Tumor response was evaluated every two cycles per Response Evaluation Criteria in Solid Tumors (RECIST) v1.1, with progression-free survival (PFS) and overall survival (OS) as endpoints. Adverse events (AEs) were graded according to National Cancer Institute-Common Terminology Criteria (NCI-CTC) 5.0. Blood samples were collected at baseline and during treatment to assess immune function, neuron-specific enolase (NSE), pro-gastrin-releasing peptide (ProGRP), neutrophil-to-lymphocyte ratio (NLR), platelet-to-lymphocyte ratio (PLR), and lactic dehydrogenase (LDH). Results: All patients received a chemotherapy regimen of 4–6 cycles of carboplatin plus etoposide (EC) or cisplatin plus etoposide (EP). In the latest efficacy assessments, 3.1% achieved complete response (CR), 90.6% achieved partial response (PR), 6.3% achieved stable disease (SD), with overall response rate (ORR) of 93.7% and disease control rate (DCR) of 100%. Adverse reactions related to treatment included grade 3/4 myelosuppression occurring in 15.6%, grade 4 thrombocytopenia in 3.12% and radiation pneumonitis in 18.7%. Follow-up through June 2025 showed median PFS of 16.3 months [95% confidence interval (CI): 11.2–21.3] and median OS of 37 months (95% CI: 30.0–44.0) for first-line treatment. The incidence of abnormal baseline immune function was 78.9%, of which 80% presented as immune suppression. PLR demonstrated significant prognostic value: patients with baseline PLR >101.75 had longer PFS. Similarly, at the end of the second cycle or before radiotherapy, patients with PLR >60.1 also showed prolonged PFS. In contrast, dynamic changes in NLR did not demonstrate prognostic significance [hazard ratio (HR) =1.05, 95% CI: 0.32–3.53, P=0.93]. LDH testing suggested that patients with normal baseline LDH had longer PFS than those with elevated levels (18.5 vs. 10.0 months), although the difference was not statistically significant (P=0.15). Notably, in some patients, LDH levels decreased from elevated to normal during treatment, suggesting a potential association with treatment response. Conclusions: Chemotherapy combined with ICIs followed by sequential or concurrent radiotherapy is feasible, safe, and effective for LS-SCLC, achieving high short-term response rates and prolonged PFS. PLR strongly predicts PFS, while normal or decreasing LDH indicates favorable treatment response. Elevated NSE and ProGRP show trends toward shorter PFS. These biomarkers can guide prognosis and therapy monitoring, supporting personalized management in LS-SCLC.

Conventional three-dimensional (3D) organoid models are limited by high cost, technical complexity, and poor replication of tumor-immune interactions. To address these limitations, we developed two novel 2.5D systems from malignant pleural effusion (MPE) of patients with lung adenocarcinoma. The first uses MPE-derived adherent cells to create matrix-free niches, while the second combines MPE supernatant with DMEM-F12 and 10-30 % fetal bovine serum, with or without agarose, to form a thermoresponsive matrix that mimics dynamic tumor microenvironments. Autologous MPE supernatants were used to preserve native cytokines and extracellular matrix components. Genomic and immune microenvironment analyses demonstrated over 90 % concordance between 2.5D organoids and parental tumors, which aligned with the standards reported for existing 3D tumor-organoid cultures. The 2.5D organoids retained key functional characteristics, including mucus secretion, phenotypic plasticity, and angiogenesis. Organoid-based high-throughput screening enabled identification of candidate therapeutics, and drug sensitivity assays correlated strongly with patient outcomes, underscoring their utility for personalized medicine. The matrix-free design facilitated rapid organoid generation (3-10 days) and reduced costs by approximately 90 % relative to conventional 3D methods. These 2.5D models provide a clinically translatable platform that replicates tumor heterogeneity while overcoming cost and technical barriers, offering a promising tool for precision pharmacology and oncology.

Background: The treatment of patients with advanced epidermal growth factor receptor (EGFR)-mutated non-small-cell lung cancer (NSCLC) whose disease progresses after tyrosine-kinase inhibitors (TKIs) treatment has become a research hotspot. Objective: To identify effective and safe treatment options for patients with EGFR-mutated advanced NSCLC who progressed on TKIs. Methods: We searched databases including PubMed, Cochrane Library, and major international conference abstracts (2018–2023) to identify phase II/III randomized controlled trials (RCTs) and single-arm studies of EGFR-mutated advanced NSCLC post-TKI progression from April 2018 to June 2024. Outcomes included progression-free survival (PFS), overall survival (OS), objective response rate (ORR), and grade ≥3 adverse events (AEs), treatment-related AEs (TRAEs), and TRAE-related deaths. Bayesian network meta-analysis and individual patient data (IPD) meta-analysis were performed to compare treatment efficacy and safety. Results: This meta-analysis included randomized controlled trials (RCTs) and 5 single-arm phase 2 trials (3116 patients) evaluating 7 treatment regimens for EGFR-mutated advanced NSCLC post-TKI progression. In the network meta-analysis (NMA), amivantamab plus lazertinib plus chemotherapy (amiva-lazer-chemo) yielded the highest PFS (surface under the cumulative ranking curve [SUCRA]: 0.88; hazard ratio [HR] vs chemotherapy, 0.44; 95% CI, 0.32-0.61), followed by AK112 plus chemotherapy (SUCRA: 0.79; HR, 0.46; 95% CI, 0.32-0.67). All regimens significantly improved PFS compared with chemotherapy alone. Amivantamab plus chemotherapy ranked highest for ORR (SUCRA: 0.82; odds ratios [OR] vs chemotherapy, 3.16; 95% CI, 1.09-9.41). Amiva-lazer-chemo had the highest grade ≥3 AE incidence. IPD analysis confirmed superior PFS for amiva-lazer-chemo (median, 8.45 months; 95% CI, 7.02-9.26; HR vs chemotherapy, 0.47; 95% CI, 0.40-0.55; P <.001). Moderate ORR heterogeneity (I² = 52.2%) and high AE heterogeneity (I² = 79.5%-92.1%) were noted. Conclusion: In this meta-analysis of patients with TKI-resistant EGFR-mutated advanced NSCLC, the amiva-lazer-chemo regimen was associated with longer PFS at both the study level and individual patient level. Combination therapy with anti-angiogenic agents also represents a viable treatment strategy for this patient population. Systematic Review Registration: https://www.crd.york.ac.uk/PROSPERO/ , identifier CRD42024565403.