Frontiers in Oncology

Background: Accurate preoperative prediction of lymph node metastasis (LNM) in papillary thyroid carcinoma (PTC) remains challenging, particularly in clinically node-negative (cN0) patients, leading to potential overtreatment. We aimed to develop and validate a Transformer-based 2.5D deep learning model (ThyLNT) using preoperative computed tomography (CT) images for robust prediction of LNM and to explore its underlying biological basis through multi-omics analyses. Methods: A total of 1,560 PTC patients from six hospitals were retrospectively included. The Tongji Hospital cohort (n=1,010) was divided into training (70%) and internal validation (30%) sets, while five independent institutions served as external test cohorts. For each lesion, seven 2.5D slices were extracted and modeled using a DenseNet201 backbone. Slice-level features were integrated using a Transformer-based feature-level fusion strategy and compared with ensemble learning, multi-instance learning (MIL), and traditional radiomics approaches. Model performance was assessed using area under the receiver operating characteristic curve (AUC), calibration analysis, decision curve analysis (DCA), and precision-recall curves. Multi-omics analyses, including bulk RNA-seq, single-cell RNA-seq, spatial transcriptomics, and spatial metabolomics, were performed to investigate biological correlates. Results: The Transformer-based model consistently outperformed comparator models across cohorts. In the training and validation cohorts, ThyLNT achieved AUCs of 0.882 and 0.787, respectively, with external AUCs ranging from 0.772 to 0.827. Compared with ultrasound (US) and CT, ThyLNT showed superior predictive performance (all P < 0.001 in the validation cohort). Simulation analysis in cN0 patients suggested that ThyLNT could reduce unnecessary lymph node dissection (LND) from 52.16% to 4.88%. Transcriptomic analysis combined with WGCNA and correlation analysis identified VEGFA as the gene most strongly associated with ThyLNT prediction scores. Single-cell and spatial transcriptomic analyses suggested metastasis-related tumor microenvironment remodeling, while enrichment analysis of genes affected by virtual knockout of VEGFA indicated involvement of angiogenesis- and epithelial-mesenchymal transition (EMT)-related pathways. Spatial metabolomics further revealed coordinated lipid metabolic reprogramming in metastatic tissues. These findings suggest that ThyLNT provides robust predictive performance while capturing biologically relevant features associated with metastatic progression.

BackgroundGastric cancer is one of the most common malignancies worldwide and is associated with poor prognosis, placing a considerable burden on public health. Overall treatment outcomes remain unsatisfactory, and accurate lymph node staging is essential for optimizing therapeutic strategies and improving survival. This study aimed to compare the prognostic value of different lymph node staging systems in patients with gastric adenocarcinoma and to provide a more refined prognostic assessment tool for clinical practice. MethodsWe included 4,054 patients with gastric adenocarcinoma from the SEER database (2015-2019) and 383 patients from the First Affiliated Hospital of Hainan Medical University. All patients underwent gastrectomy with D2 lymphadenectomy. Clinicopathological variables included sex, age, race, tumor size, T stage, AJCC N stage (AJCC-N), lymph node ratio (LNR), and log odds of positive lymph nodes (LODDs). Between-group comparisons were performed using the chi-square test. Optimal cut-off values were determined with X-tile software. Survival differences were evaluated by Kaplan-Meier curves. Receiver operating characteristic (ROC) curves were used to compare predictive performance. Cox regression models were applied to identify independent prognostic factors, which were then incorporated into a nomogram. Model performance was assessed using calibration curves and decision curve analysis (DCA). ResultsAJCC-N, LNR and LODDs were strongly and positively correlated in all three datasets (P < 0.001). ROC analysis showed that LODDs had slightly larger areas under the curve than LNR and AJCC-N for predicting 1-, 3- and 5-year survival. Multivariable Cox regression confirmed that LODDs, together with sex, age, race, T stage and tumor size, were independent risk factors for overall survival (P < 0.05). The nomogram constructed from these factors showed good agreement between predicted and observed outcomes on calibration curves, and DCA indicated meaningful clinical net benefit across a broad range of threshold probabilities. ConclusionBy integrating the numbers of positive and negative lymph nodes, LODDs more sensitively reflects changes in metastatic tumor burden and showed the best prognostic performance among the evaluated systems for gastric adenocarcinoma. The proposed nomogram may serve as a useful tool for individualized prognostic assessment.

Purpose This study aimed to evaluate the efficacy and safety of neoadjuvant chemotherapy (NAC) combined with the programmed death protein 1 (PD-1) inhibitor sintilimab versus NAC alone in patients with triple-negative breast cancer (TNBC). Materials and Methods In this retrospective cohort study, we collected clinical data from 61 patients with triple-negative breast cancer (TNBC) who received neoadjuvant therapy at The First Hospital of Lanzhou University between July 2024 and July 2025. These patients were divided into two groups: the neoadjuvant chemotherapy (NAC) plus sintilimab group (n=27) and the NAC-alone group (n=34). The primary endpoint was the pathological complete response (pCR) rate. Secondary endpoints included objective response rate (ORR), safety, and changes in tumor markers. Results The combination therapy group showed significantly higher ORR (85.2% vs. 58.8%) and pCR rates (59.3% vs. 32.4%) compared to the NAC alone group (both P<0.05). Post-treatment Ki-67 levels were also significantly lower in the combination group (P<0.05). The overall incidence of adverse events was comparable between groups (P>0.05), although leukopenia was more frequent with sintilimab (P<0.05). Conclusion In the neoadjuvant setting for TNBC, the addition of sintilimab to NAC significantly improves ORR and pCR rates, effectively reduces the tumor proliferation index Ki-67, and does not significantly increase the overall burden of adverse events. The combination regimen shows a manageable safety profile and demonstrates positive clinical value. Keywords Triple Negative Breast Cancer, Immunotherapy, Sintilimab, Combination neoadjuvant chemotherapy, Efficacy, Real-World data.

Background: To investigate the safety and efficacy of CTguided lung nodule localization needles for the preoperative localization of small pulmonary nodules. Methods: A retrospective study was conducted on 102 patients with a total of 113 small pulmonary nodules who underwent preoperative localization at Jinan Fourth People's Hospital from January 2024 to December 2025. Nodule diameter and depth, localization time, the number of pleural punctures, the localization success rate, and postoperative complications (hook dislodgement, hemorrhage, and pneumothorax) were recorded. All patients underwent video assisted thoracoscopic surgery (VATS) after localization. Results: The mean nodule diameter was 0.97{+/-}0.36 cm, the mean depth was 1.26{+/-}0.48 cm, and the mean localization time was 9.8{+/-}3.65 minutes. The hook dislodgement rate was 0.98% (1/102), the intrapulmonary hemorrhage rate was 14.71% (15/102), and the pneumothorax rate was 16.67% (17/102). All pulmonary nodules were successfully resected by VATS at 73.82{+/-}13.83 minutes after localization, and no severe complications occurred. Conclusions: The use of a CTguided lung nodule localization needle for the preoperative localization of small pulmonary nodules decreases the time needed for intraoperative nodule detection and operation time. This strategy is a simple, safe, and accurate preoperative localization method that is worthy of increased clinical use.

IntroductionColorectal cancer (CRC) remains a leading cause of cancer-related morbidity and mortality worldwide, despite advances in conventional oncological therapies. In recent years, various studies have made advances in integrative oncology, such as investigating the use of Chinese Herbal Medicine (CHM) as a complementary therapy alongside conventional oncological therapies to alleviate treatment-related adverse effects, improve quality of life, and potentially enhance therapeutic outcomes. Despite this, clinical practice in this area remains highly heterogeneous, with limited standardized guidelines on key areas of concern such as (1) optimal intervention, (2) recommended stage and duration of intervention, (3) safety considerations, and (4) possible herb-drug interactions. Hence, this study aims to establish expert consensus on the usage of CHM as a complementary therapy in the management of CRC, to support safe, consistent, and evidence-informed clinical practice. Methods and AnalysisWe will employ a modified Delphi technique to achieve consensus amongst a panel of international experts in various fields related to integrative oncology. Prior to the study, a list of questionnaire items was developed based on a systematic review of existing clinical practice guidelines on CRC. An international panel will be invited based on established international profile in integrative oncology research and clinical practice, and by peer referral. Two rounds of Delphi will be conducted using anonymous online questionnaires. Consensus will be considered reached if at least 50% of the panel strongly agree/disagree that an item should be included or excluded while strong consensus will be set at 76%. Items which achieve strong consensus after Round 1 will be removed, before being sent out for Round 2 with a summary of Round 1 responses for a final consensus. Ethics and DisseminationEthics approval has been obtained from the Institutional Review Board of Nanyang Technological University (IRB-2025-1222). Our findings will be disseminated through peer-reviewed publications and conference presentations. Strengths and limitations of this studyO_LIThis study will develop an expert consensus which aims to guide future integration of Chinese Herbal Medicine (CHM) as a complementary therapy into colorectal cancer (CRC) management. C_LIO_LIKey concerns in areas such as determining the (1) optimal intervention, (2) recommended stage and duration of intervention, (3) safety considerations, and (4) possible herb-drug interactions, thereby laying the groundwork for potential future incorporation of CHM into CRC treatment protocols alongside conventional oncology approaches has been identified, thus limiting implementation in clinical practice. C_LIO_LIDesigning a study e-guide, followed by the consensus rounds study online will facilitate participants responses and the dissemination of information from previous rounds. C_LI

Background and ObjectiveDespite its prevalence, bladder cancer (BC) remains an unsolved pathogenesis. It is believed that TRPC6 channels have unique electrophysiological properties that contribute to intracellular Ca2+ signalling and tumorigenesis in cells. However, the mechanism by which TRPC6 contributes to BC progression and intracellular Ca2+ homeostasis remains unclear. MethodIn this study, TRPC6 expressions in paired BC and adjacent normal tissues were measured by immunohistochemistry. A KEGG pathway enrichment analysis was conducted to determine TRPC6s potential contribution to BC. Ca2+ imaging analysis was performed to explore the contribution of TRPC6 in the BC cell. Flow cytometry and Cell Counting Kit-8 assay were performed to explore the effects of TRPC6 on the proliferation of BC. The impacts of TRPC6 SOCE on PI3K/Akt/mTOR pathway were measured by western blot. Based on the above bunch of studies, TRPC6 was found to be overexpressed in human BC tissue, which correlated with poor survival rates for patient overall survival (OS). We used the TRPC6-specific antagonist SAR7334 to explore and reveal significant inhibition of BC cell proliferation. Mechanistically, TRPC6 mediated cytosolic Ca2+ and regulation of SOCE, leading to the activation of the IP3K/AKT/mTOR pathway. ResultsWe found that SAR7334 arrested the phosphorylation of PI3K and Akt, thus causing a significant decrease in phosphorylated mTOR. Similar effects were observed for the SOCE-specific antagonist MRS1845. In contrast, the Akt inhibitor MK2206 did not alter the SOCE in BC cells. Conclusionsour results indicate that the pharmacological inhibition of TRPC6 arrests tumour cell proliferation through SOCE targeting the PI3K/Akt/mTOR pathway. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=101 SRC="FIGDIR/small/702689v1_ufig1.gif" ALT="Figure 1"> View larger version (28K): org.highwire.dtl.DTLVardef@bf783aorg.highwire.dtl.DTLVardef@6e3487org.highwire.dtl.DTLVardef@13aad23org.highwire.dtl.DTLVardef@13cfe7e_HPS_FORMAT_FIGEXP M_FIG C_FIG Schematic of proposed pathway. A model for summarizing TRPC6 contributes to cell proliferation in bladder cancer. TRPC6 specific inhibitor SAR7334 alter cellular cytosolic Ca2+ and SOCE, thus arrested phosphorylation of the PI3K/Akt pathway and the proliferation of bladder cancer cells.

BackgroundCyclin-dependent kinases drive the progression through the cell cycle and thereby form classical targets for cancer therapy. In prostate cancer (PC), the first line of therapy typically targets androgen receptor (AR), but it frequently leads to development of incurable form of the disease, castration-resistant PC (CRPC). Here, we sought to understand if CRPC cells are selectively addicted to a specific cell cycle kinase. MethodsWe used PC and CRPC patient data to evaluate transcriptional changes and modeled the responses in vitro using multiple models of PC, CRPC and normal cells. Development of a CDK2 inhibitor-resistant CRPC cell line, and a compound screen were used to identify chronic and acute vulnerabilities to augment the efficacy of our candidate therapy in multiple PC, CRPC and also normal cells, to assure selectivity. ResultsWe show that the emergence of CRPC is associated with significant upregulation of cyclins that positively regulate cyclin-dependent kinase 2 (CDK2) and downregulation of CDK4 cyclins. Accordingly, CDK2-specific inhibitors and its knock down efficiently reduce proliferation of PC and CRPC cells. CDK2 inhibitor-resistant CRPC model displayed transcriptional rewiring of cell cycle regulators, characterized by a shift towards CDK4/6-dependency and increased AR-signaling. Combinatorial drug screen discovered both antagonistic and additive combinations, and we show that AR inhibitors selectively augment the efficacy of CDK2 inhibitors against PC and CRPC cells, but the combination is not toxic to normal cells. ConclusionWe discovered that CRPC cells are addicted to high CDK2 activity and show that combination of CDK2 inhibitors with the currently used anti-CRPC therapies selectively augment their efficacy.

Background and AimsA combination of chemical carcinogens and Western diet have been used to induce hepatocellular carcinoma (HCC) in mice, but these models show low incidence of HCC in female mice, while mice that do develop HCC show a wide range of HCC stage development. Therefore, a more reliable mouse model that induces advanced stage HCC in both male and female mice is warranted, which will enable reliable preclinical testing of therapeutics for advanced HCC. The purpose of this study was to create a simple, yet tolerable and reliable chemical carcinogen and Western diet induced mouse model of advanced stage HCC for both male and female mice. Approach and ResultsWe report that providing mice with a Western diet at weaning and for their lifetime combined with the sequential administration of low dose diethylnitrosamine (DEN), thioacetamide (TAA), and sucrose water promotes stage 2-3 HCC by 30 weeks of age in 100% of male mice and 96% of female mice (4% had stage 1 HCC), with a low fatality rate. Spatial transcriptomics, proteomics, and western blotting revealed this model alters genes and proteins similar to human HCC. ConclusionsThis study introduces, for the first time, a reliable chemical carcinogen model that induces advanced HCC in both male and female mice. Importantly, the model was tolerable for mice and induced protein and gene signatures comparable to human HCC. This new protocol will be a valuable model for preclinical testing of new therapeutic approaches for advanced HCC.

PurposeTreatment options of advanced oral squamous cell carcinomas (OSCC) are limited, and cisplatin toxicity and drug resistance are major clinical issues. Src is a central kinase that integrates multiple oncogenic pathways and a promising therapeutic target. However, Src inhibitors have shown suboptimal efficacy as monotherapies and their sensitivity in OSCC remains elusive. Experimental DesignWe examined the activation of major oncogenic signaling pathways and the antitumor effects of six Src inhibitors (dasatinib, ponatinib, vandetanib, saracatinib, PP2, bosutinib) in seven human OSCC cell lines (HSC-2, HSC-3, HSC-4, SAS, HO-1-u-1, CAL27, SCC-25). BALB/cAJcl nu/nu mice bearing CAL27 xenografts received dasatinib (30 mg/kg, intraperitoneally, daily), bosutinib (50 mg/kg, intraperitoneally, daily), cisplatin (2 mg/kg or 4 mg/kg, intraperitoneally, weekly), or combinations. Tumor volume, bioluminescence imaging, and body weight were monitored for 17 or 21 days, followed by histopathological assessment. ResultsThe activation of the key pathways, including Src and MAPK, considerably differed among the cell lines and was linked to heterogeneous sensitivity to Src inhibitors. Effective growth suppression required Src dephosphorylation and downstream MAPK pathway inhibition, which vary depending on the cell line. Additionally, combination treatment with a Src inhibitor and cisplatin showed additive antitumor effects, allowing the reduction of cisplatin doses by half without efficacy loss. Notably, dasatinib alone and in combination with cisplatin decreased tumor burden with characteristic internal tumor death in vivo. ConclusionsThese findings elucidate Src signaling dependency on OSCC and the potential of Src inhibition to decrease cisplatin toxicity, paving way for Src targeted therapeutic strategies.

Immune checkpoint inhibitors such as anti-PD1 antibodies are essential in cancer therapy. Emerging data suggest that lower doses may be effective and more economical, though further evidence is needed. We conducted a retrospective study at Centre Leon Berard to assess the efficacy and safety of low-dose nivolumab (20 mg every three weeks) in patients with advanced cancer, mainly squamous cell carcinomas (SCC). Between 2023 and 2024, 53 patients were treated, with a median age of 74 years; 39.6% were over 80. Most were male (64%) and had ECOG >1 (69.9%). Primary tumor sites included cutaneous SCC (34%), head and neck SCC (32%), and soft tissue sarcoma (15%). After a median follow-up of 8.3 months, median overall survival was 7.5 months. The objective response rate (ORR) was 20.8% overall, rising to 35.3% in cutaneous SCC and 23.5% in head and neck SCC-comparable to standard-dose nivolumab. Toxicity was manageable: 18.7% experienced immune-related adverse events, with only 3.7% grade 3. Low-dose nivolumab demonstrates encouraging efficacy and tolerability in a frail population, supporting its potential role in resource-limited settings. Prospective trials are warranted to confirm these findings in broader populations.

PurposeIntravesical interferon-alpha (IFN) gene therapy has shown promise in treating BCG-unresponsive non-muscle invasive bladder cancer (NMIBC). Ongoing work in our lab aims to further improve its treatment efficacy by identifying resistance mechanisms and deploying targeted combination treatment strategies. Experimental designWe performed end-tumor RNA-seq analysis of MB49 murine tumors treated with IFN gene therapy, identifying the ErbB pathway as a resistance mechanism. We consequently hypothesized that a combination treatment involving an ErbB pathway blocker and IFN could yield improved outcomes. MB49 cells were treated in vitro with lentiviral IFN (LV-IFN) gene therapy, with/without Afatinib, a pan-ErbB inhibitor, and cell viability and migration assays were performed. Next, in vivo studies were conducted in the syngeneic MB49 orthotopic murine bladder cancer model. The mice were randomized into 5 treatment groups (n=10 each): saline (Ctrl), LV-Ctrl, oral Afatinib monotherapy, intravesical LV-IFN monotherapy, and the experimental intravesical LV-IFN + oral Afatinib combination therapy. Overall survival (OS) and drug toxicity were assessed. ResultsCombination therapy significantly reduced MB49 cell viability in vitro compared to all other treatment conditions (mean relative ATPase activity at 72 h for the combination treatment was 4%, compared to 100%, 26%, and 28% for Ctrl, LV-IFN, and Afatinib, respectively, p<0.001). This additive effect on cell viability appeared to be driven by a combination of early-cytostatic and late-cytolytic effects. The combination treatment also markedly inhibited cell migration (mean migrated cells/10x Boyden chamber assay at 36 h were: 92.3 for the combination therapy and 631.0, 600.4, and 270.3 for Ctrl, LV-IFN, and Afatinib, respectively, p<0.001). Finally, the in vivo studies demonstrated improved OS with combination therapy (median OS was 49 d in the combination group vs 15, 29, and 26 d in Ctrl, LV-IFN, and Afatinib groups, respectively, Log-rank p<0.001). No mice in the combination therapy group died of drug toxicity. ConclusionsOur preliminary findings suggest that the ErbB pathway may serve as a clinically significant resistance mechanism to intravesical IFN gene therapy, and when targeted concurrently, may improve treatment efficacy.

Malignant peripheral nerve sheath tumors (MPNSTs) are highly aggressive sarcomas with poor prognosis and a strong tendency for metastasis and relapse. Surgical removal remains the mainstay of treatment but is frequently ineffective or impractical. Currently, no effective targeted therapy exists for this type of malignancy. PRMT5 has recently emerged as a promising therapeutic target in various human cancers with MTAP loss, which results in cancer cell dependency on PRMT5 activity. The frequent loss of MTAP in MPNSTs suggests that PRMT5 inhibition is a promising therapeutic option and enables the stratification of cancer patients with few treatment options. We first examined human nerve sheath tumor samples and found that increased PRMT5 expression and activity correlated with MTAP loss in 86.8% (33/38) of MPNSTs and in atypical neurofibromatous neoplasm with uncertain biologic potential (ANNUBP) (5/5). When PRMT5 activity was inhibited genetically and chemically, the cell growth of MTAP-deficient MPNST cell lines was suppressed, but not that of MTAP-proficient MPNST cell lines. Moreover, in the PRMT5-inhibited MTAP-deficient MPNST cell lines, spontaneous DNA damage accumulation was observed following G2/M cell cycle arrest. The DNA replication stress marker RPA32 decreased, and CHK1 was activated early after PRMT5 knockdown, likely contributing to the accumulation of DNA damage. In addition, we combined PRMT5 inhibition with the DNA-damaging agents doxorubicin and gemcitabine, resulting in synergistic effects and increased cancer cell death in MTAP-deficient MPNST cell lines. Together, these findings identify PRMT5 as a compelling therapeutic target in MTAP-deficient MPNSTs. This PRMT5 inhibition strategy has strong translational potential for MPNSTs. GRAPHICAL ABSTRACT O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=181 SRC="FIGDIR/small/710638v1_ufig1.gif" ALT="Figure 1"> View larger version (33K): org.highwire.dtl.DTLVardef@15abc35org.highwire.dtl.DTLVardef@1fa4ebborg.highwire.dtl.DTLVardef@470c51org.highwire.dtl.DTLVardef@79cc3f_HPS_FORMAT_FIGEXP M_FIG C_FIG

ObjectiveThe purpose of the present study is to describe the survival outcomes of patients with low-grade serous ovarian cancer (LGSOC) in the post-operative setting from a tertiary gynecologic oncology referral centre in Quebec, including evaluation of patient characteristics, clinical outcomes and prognostic factors. MethodsThe study included 25 patients: 1) with a post-surgical histopathologic diagnosis of a low-grade serous tumour of the ovary, 2) underwent primary cytoreductive surgery prior to adjuvant therapy, and 3) for whom clinical data was available. Clinical and demographic features were characterized by descriptive statistics. Clinical endpoints of progression-free survival (PFS) and overall survival (OS) were assessed, utilizing the Kaplan-Meier method for estimating survival probabilities. ResultsThe median age of this cohort was 61 years (range, 26-81). Median OS was 140.6 months in patients with no residual disease (R0), 71 months in patients with microscopic residual disease (R1), and 27.7 months in patients with macroscopic residual disease (R2) (p=.001). Residual disease was also found to significantly impact PFS (p=.008). Administration of adjuvant chemotherapy failed to improve survival outcomes altogether (PFS, p = .270; OS, p = .300). ConclusionsThis study supports the shifting consensus that optimal cytoreductive surgery, where feasible, is paramount for successful treatment of LGSOC. Furthermore, treatment with adjuvant chemotherapy may lead to worse survival outcomes.

BackgroundOlder age and comorbidities complicate initial therapy in non-small-cell lung cancer (NSCLC), as platinum ineligibility has not been systematically characterized. MethodsAll 2592 patients presenting with metastatic NSCLC between 2018-2023 at Thoraxklinik Heidelberg were analyzed. ECOG status (PS), comorbidities, molecular testing, therapy, toxicities, and outcomes were verified from individual patient records. ResultsAmong 1306 patients with PD-L1 0-49%, systemic therapy was initiated in 74%. With availability of monoimmunotherapy, the treatment rate for patients with PD-L1[&ge;]50% (n=507) was higher by 5% (p=0.01), while best supportive care (BSC) by own choice was reduced (1.8% vs. 4.5%, p=0.005) more than medical BSC (mBSC 14.6% vs. 17.8%, p=0.11), and early death remained unchanged (ca. 4%). Initial suitability for systemic therapy was documented for 70% of cases eventually receiving mBSC after deterioration associated with comorbidities, metastatic burden, longer workup duration, or radiotherapy upfront (all p<0.001). The atezolizumab Summary of Medicinal Product Characteristics (SmPC) criteria, i.e. >80 years, or PS [&ge;]3, or comorbidities with PS [&ge;]2 or with age [&ge;]70, were fulfilled by 38% of patients (n=501) and associated with a >3-fold higher risk of BSC or early death (230/501), as well as significantly higher toxicity under platinum and shorter survival, which for a platinum dose ratio [&le;]60% across 4 cycles (9% of 1306) was similar to that with single-agent chemotherapy (median 5.1 months, p<0.001). SmPC criteria correlated better than comorbidity scores with foregoing platinum, but predictive performance for individual patients remained modest (AUC 0.71, p<0.001). ConclusionsThe high initial attrition of approximately 25% in NSCLC could improve with availability of monoimmunotherapy, but requires optimized, faster patient workflows for better mitigation. Adoption of the SmPC criteria could support a priori identification of patients at risk for mBSC or platinum overtreatment to enhance utilization of monoimmunotherapy and other novel platinum-free first-line options in the future. HighlightsO_LIA high initial attrition of approximately 25% is caused by deterioration after histologic diagnosis in advanced NSCLC. C_LIO_LIMonoimmunotherapy and optimized workflows may facilitate treatment for ca. 15% additional stage IV NSCLC patients. C_LIO_LISmPC criteria indicate cases at higher risk for BSC (>3x) or platinum overtreatment (i.e. platinum dose ratio [&le;]60%). C_LIO_LISmPC patients receiving platinum have higher toxicity and shorter survival than non-SmPC patients. C_LIO_LIImproved therapeutic allocation will be essential for utilization of any novel platinum-free option in the future. C_LI

Bladder cancer (BC), particularly muscle-invasive and metastatic disease, remains a major clinical challenge despite recent advances in immunotherapy. In this study, we aimed to identify a promising antitumor compound from five candidate small molecules and to explore its potential roles in BC progression. Through antiproliferative screening, cryptotanshinone (CTS) was identified as the promising candidate. Using both two-dimensional BC cell lines and three-dimensional bladder tumor organoid models, we comprehensively evaluated the effects of CTS on cell proliferation, migration, apoptosis, and organoid growth. To further explore the underlying mechanisms, transcriptomic sequencing based on bladder cancer organoid models, protein-protein interaction network analysis, and public databases (TCGA-BLCA, TIMER, and TISIDB) were integrated to examine immune-related pathways and potential molecular targets associated with CTS. GeneMANIA network prediction and molecular docking analyses were subsequently performed to investigate upstream regulatory networks and the potential interactions between CTS and key components of the cGAS-STING-IFN-I-JAK-STAT signaling pathway. Integrative analyses suggested that IFIT1, IFIT2, and IFIT3 may function as immune-associated genes potentially linked to BC progression, patient prognosis, immune cell infiltration, and PD-1/PD-L1 expression. Molecular docking results suggested that CTS may interact with core regulatory proteins within the cGAS-STING-IFN-I-JAK-STAT pathway, potentially influencing IFIT transcriptional regulation. Collectively, these findings indicate that CTS exhibits measurable antitumor and immunomodulatory effects, which may be associated with modulation of the cGAS-STING-IFN-I-JAK-STAT-IFIT signaling axis, supporting its potential as a small-molecule candidate for bladder cancer.

BackgroundPancreatic cancer is a leading cause of cancer mortality, and early recognition is challenging. To achieve early diagnosis using symptoms alone, we examined patterns across different stages using network analysis to derive clinically useful insights. MethodsSymptom variables from a de-identified dataset of 50,000 pancreatic cancer patients were analyzed. Stratification by stage was done, followed by bootstrap resampling to address imbalances across strata. Symptom networks were then constructed with nodes representing symptoms and edges representing conditional dependencies estimated via an Ising-style neighborhood selection approach implemented through L1-regularized logistic regression. Strength, betweenness, and closeness centrality indices were then calculated, and their stability was analyzed using the case-dropping bootstrap. Network comparison tests were done, and difference networks were analyzed. Spring-layout algorithm was used for visualization, with node size being the predictability (pseudo-R{superscript 2}), and the edge weight being the mean partial correlation magnitude. ResultsOn average, symptoms were present in about one out of four patients (M = 0.26). Weight loss and abdominal discomfort were the most prevalent of the symptoms, followed by jaundice and back pain. Network structures became sparser across stages with a decreasing number of edges and centrality indices. Jaundice emerged as the dominant hub in Stage I, but shared dominance with Weight Loss in Stage II. Node predictability (pseudo-R2) was effectively zero across all disease stages. ConclusionOur network analysis of pancreatic cancer symptomatology across stages revealed distinct patterns that may improve understanding of its clinical presentation and support earlier recognition.

Patients with invasive lobular carcinoma of the breast (ILC) are at high risk of long-term recurrence and metastatic progression with poor prognoses due to delayed detection and treatment-refractory disease. Unfortunately, few models are available to investigate metastatic ILC (mILC) and understand the unique metastatic patterns and phenotypes, including abdominal metastases, leptomeningeal disease, and mixed osteosclerotic/lytic bone metastases. Therefore, we expanded upon the previously established mammary intraductal (MIND) cell line xenograft model by supplementing mice with low-dose estradiol to promote disease progression. We observed spontaneous multi-organ spread from the mammary gland to common and mILC-specific tissues, with micro-metastatic disease as early as 12 weeks post-engraftment and macro-metastatic disease in 24-30 weeks, without the need for primary tumor resection. Primary and metastatic tumors remain highly endocrine responsive, allowing for the evaluation of novel therapeutics in the setting of disseminated metastasis. Derivative cell lines were isolated from various metastatic lesions, a total of 13 derivates from 7 sites across three hosts, and were found to have shared gene expression changes related to metabolism and intercellular signaling. Focusing on bone-derived variant cells as bone is the most common site for mILC to present, we found that bone-derived variant lines maintain multi-organ metastatic potential upon rechallenge by MIND or intratibial injection, despite increased aggressiveness and maintained endocrine response. Notably, bone lesions from either challenge route showed mixed osteosclerotic/lytic features characteristic to clinical ILC. Accordingly, we found that conditioned medium from ILC cells and the mILC bone-derived variants induce osteoblast differentiation and suppressed osteoclast differentiation in vitro, consistent with their effect on bone remodeling in vivo and in clinical disease. Together, the models developed herein can be utilized to understand the unique metastatic processes of mILC, and to investigate new therapeutic combinations in the setting of endocrine-responsive primary and metastatic ILC.

IntroductionManual data extraction from unstructured clinical notes is labor-intensive and impractical for large-scale clinical and research operations. Existing automated approaches typically require large language models, dedicated computational infrastructure, and/or task-specific fine-tuning that depends on curated data. The objective of this study is to enable accurate extraction with smaller locally deployed models using a disease-site specific pipeline and prompt configuration that are optimized and reusable. Materials/MethodsWe developed OncoRAG, a four-phase pipeline that (1) generates feature-specific search terms via ontology enrichment, (2) constructs a clinical knowledge graph from notes using biomedical named entity recognition, (3) retrieves relevant context using graph-diffusion reranking, and (4) extracts features via structured prompts. We ran OncoRAG using Microsoft Phi-3-medium-instruct (14B parameters), a mid-size language model deployed locally via Ollama. The pipeline was applied to three cohorts: triple-negative breast cancer (TNBC; npatients=104, nfeatures=42; primary development), recurrent high-grade glioma (RiCi; npatients=191, nfeatures=19; cross-lingual validation in German), and MIMIC-IV (npatients=100, nfeatures=10; external testing). Downstream task utility was assessed by comparing survival models for 3-year progression-free survival built from automatically extracted versus manually curated features. ResultsThe pipeline achieved mean F1 scores of 0.80 {+/-} 0.07 (TNBC; npatients=44, nfeatures=42), 0.79 {+/-} 0.12 (RiCi; npatients=61, nfeatures=19), and 0.84 {+/-} 0.06 (MIMIC-IV; npatients=100, nfeatures=10) on test sets under the automatic configuration. Compared to direct LLM prompting and naive RAG baselines, OncoRAG improved the mean F1-score by 0.19 to 0.22 and 0.17 to 0.19, respectively. Manual configuration refinement further improved the F1-score to 0.83 (TNBC) and 0.81 (RiCi), with no change in MIMIC-IV. Extraction time averaged 1.7-1.9 seconds per feature with the 14B model. Substituting a smaller 3.8B model reduced extraction time by 57%, with a decrease in F1-score (0.03-0.10). For TNBC, the extraction time was reduced from approximately two weeks of manual abstraction to under 2.5 hours. In an exploratory survival analysis, models using automatically extracted features showed a comparable C-index to those with manual curation (0.77 vs 0.76; 12 events). ConclusionsOncoRAG, deployed locally using a mid-size language model, achieved accurate feature extraction from multilingual oncology notes without fine-tuning. It was validated against manual extraction for both retrieval accuracy and survival model development. This locally deployable approach, which requires no external data sharing, addresses a critical bottleneck in scalable oncology research. Graphical abstract O_FIG O_LINKSMALLFIG WIDTH=200 HEIGHT=89 SRC="FIGDIR/small/26347717v1_ufig1.gif" ALT="Figure 1000"> View larger version (23K): org.highwire.dtl.DTLVardef@178a4e8org.highwire.dtl.DTLVardef@1928b7corg.highwire.dtl.DTLVardef@38f36org.highwire.dtl.DTLVardef@1af4d51_HPS_FORMAT_FIGEXP M_FIG C_FIG

Epidemiological studies suggest that vegetarian diets are associated with lower cancer incidence and mortality, an effect attributed in part to phytochemicals such as polyphenols and carotenoids. Although numerous in vitro experiments and investigations using immunodeficient rodent models report tumor-suppressive activities of phytochemicals, their impact on tumor progression in immunocompetent hosts remains insufficiently understood. Here, we examined the influence of a defined plant phytochemical mixture (PPM) on the growth of colon cancer liver metastases, both in vitro and in immunocompetent mice. Consistent with the prevailing literature, treatment of the murine colon cancer cell line MC38 with the PPM significantly reduced cell proliferation and survival in vitro. Strikingly, however, administration of the PPM to mice bearing MC38-derived hepatic metastases markedly accelerated tumor growth. Immunohistochemical analyses revealed a significantly increased accumulation of immune cells--specifically CD45 leukocytes and F4/80 macrophages--at the periphery of the metastatic lesions in PPM-treated animals. To assess the functional relevance of this inflammatory response, the PPM was combined with the anti-inflammatory drug prednisolone. This intervention resulted in significantly reduced metastatic burden, supporting the notion that the PPM exacerbates tumor progression through enhanced peritumoral inflammation. These findings highlight the importance of validating observations from cell culture and immunodeficient models in fully immunocompetent systems. They further emphasize that the immunomodulatory effects of plant phytochemicals warrant careful and comprehensive investigation.

Immunotherapy with immune checkpoint inhibitors and immunotherapy combined with chemotherapy have represented promising treatments for NSCLC patients leading to prolonged survival. However, the majority of patients with advanced NSCLC have a poor prognosis. The identification and development of biomarkers for stratifying responders and non responders to immune checkpoint inhibitors contribute to unravel the mechanism of immune checkpoint pathway and the immune tumor interaction underlying the responses and are urgently needed to improve clinical outcomes of immune checkpoint inhibitor treatment. In this study, we analyzed the clinical and gene mutation data of NCSLC patients treated with nivolumab containing immunotherapy or nivolumab containing immunotherapy combined with chemotherapy (the immunotherapy treated group, n=119) and chemotherapy alone (the chemotherapy alone treated group, n=991) extracted from the MSK CHORD dataset. A DeevSurv model, a deep learning based extension of the Cox proportional hazards model was trained to generate survival risk score of each patient with binary statuses of thirty one gene mutations as input features into the model. The thirty one genes were selected based on population level mutation frequency, patient level variance in mutation status, and univariate Cox proportional hazards analyses evaluating the association between the presence or absence of each gene mutation and overall survival. The performance of the trained DeepSurv model was evaluated on the test set of the immunotherapy treated group using the concordance indexes (C index). The trained model was subsequently applied without retraining to the entire chemotherapy alone treated group as a control. The resulting C indexes for the immunotherapy treated group and chemotherapy alone treated group were 0.789 and 0.483, respectively. All patients within each group were divided into high and low risk groups according to the median predicted risk score. Kaplan Meier survival curves of high and low risk groups (n=43 vs n=70) in the immunotherapy treated group revealed a significant separation (log rank p<0.001), whereas no separation was observed in chemotherapy alone treated group (p=0.62). In the combined cohort of the immunotherapy treated group and chemotherapy alone treated group, the interaction between the DeepSurv derived risk score and treatment modality was significant (HR for interaction 1.47, 95% CI from 1.32 to 1.65, p<0.005), suggesting the DeepSurv derived risk score predictive value specific to the immunotherapy. Principal component analysis and permutation importance analysis were performed as complementary analyses to assess individual genes associated with the DeepSurv derived risk score and identified ZFHX3, SMARCA4, ALK, BTK, and NOTCH2 as major contributors to survival risk stratification. Collectively. we suggested that nonlinear coupling pattern of 31 tumor gene mutation statuses in the DeepSurv model captures the heterogeneity of survival risk among nivolumab containing immunotherapy or nivolumab containing immunotherapy combined with chemotherapy treated patients with NSCLC which was visualized as clear separation between high risk and low risk groups divided by the median value of the risk scores.