4009 Background: The addition of immune checkpoint inhibitors (ICIs) to chemotherapy (chemo) in the neoadjuvant/adjuvant setting for gastric cancer (GC) has yielded mixed efficacy. Target population and treatment regimen remain to be optimized. This study compared the efficacy of serplulimab (anti-PD-1 antibody) plus chemo versus placebo plus chemo as neoadjuvant therapy and serplulimab versus chemo as adjuvant therapy for resectable GC. Methods: Patients with PD-L1 positive (PD-L1 CPS ≥ 5), histologically confirmed, untreated, resectable gastric or gastroesophageal junction (GEJ) adenocarcinoma were randomized 1:1 to receive 3 cycles of neoadjuvant intravenous (iv) serplulimab (4.5 mg/kg) or placebo, plus chemo (iv oxaliplatin, 130 mg/m 2 and oral S-1, 40-60 mg based on body surface area), followed by post-surgery adjuvant serplulimab monotherapy (up to 17 cycles) or adjuvant chemo (5 cycles), Q3W. The primary endpoint was investigator (INV)-assessed event-free survival (EFS). Efficacy superiority was first evaluated and established in patients with PD-L1 CPS ≥ 10 followed by that in the PD-L1 CPS ≥ 5 population. Results: Between November 26, 2019 and April 19, 2024, 588 patients were enrolled across 57 sites, with 292 randomized to the serplulimab group and 296 to the placebo group. As of the data cutoff date of August 19, 2025 with a median follow-up duration of 35.9 months, INV-assessed median EFS was significantly longer in the serplulimab group (n = 193) than in the placebo group (n = 217) in the PD-L1 CPS ≥ 10 population (not reached [NR], 95% CI 43.0–not estimable [NE] vs. 42.0 months, 95% CI 20.5–NE; HR 0.65, 95% CI 0.47–0.90; p = 0.0082), and in the PD-L1 CPS ≥ 5 population (NR, 95% CI 37.9–NE vs. 35.9 months, 95% CI 21.3–52.0; HR 0.73, 95% CI 0.56–0.94; p = 0.0152), meeting the protocol-specified superior criteria. Blinded independent central review-assessed median EFS was consistent with that of the investigator. Pathological complete response rate was higher in the serplulimab group than in the placebo group (21.6% vs 6.4%; odds ratio 3.95). Median OS was immature in both groups. Grade ≥ 3 treatment-related adverse events (TRAEs) were reported in 136 (46.6%) and 172 (58.5%) patients in the serplulimab and placebo groups, respectively. Discontinuation of any component of the study regimen due to TRAEs occurred in 19 (6.5%) and 31 (10.5%) patients in the respective groups. Conclusions: Neoadjuvant serplulimab plus chemo, followed by adjuvant serplulimab monotherapy, significantly prolonged EFS in patients with PD-L1-positive, resectable GC or GEJ adenocarcinoma. Improvements in other efficacy endpoints were also observed along with a favorable safety profile compared to neoadjuvant/adjuvant chemo. This world-first chemotherapy-free regimen (adjuvant) represents a promising treatment option for this indication. Clinical trial information: NCT04139135 .

Colorectal cancer (CRC) is one of the most prevalent malignancies globally and poses a substantial threat to human health. The current understanding of the biological significance of Splicing Factor 3a Subunit 2(SF3A2) in CRC remains limited. In this study, the upregulation of SF3A2 in CRC was identified through TMT-based quantitative proteomic screening of surgically resected paired primary cancer and normal epithelial tissues. Patients with elevated SF3A2 expression exhibited reduced survival compared to those with lower expression levels. Knockdown of SF3A2 significantly decreased cell proliferation, migration, and invasive properties both in vivo and in vitro. Bioinformatics enrichment analysis revealed that SF3A2 was involved in alternative splicing and functioned as an oncogene by modulating the expression of genes associated with critical tumorigenesis pathways and functions. Furthermore, immune infiltration analysis using multiple algorithms (including TIMER, EPIC, QUANTISEQ, and MCPCOUNTER) indicated an inverse relationship between SF3A2 expression levels and the presence of various immune cell types. Concurrently, predictions derived from the TIDE algorithm corroborated that patients exhibiting elevated SF3A2 expression were likely to experience a diminished response to immunotherapy. In summary, SF3A2 emerged as a promising therapeutic target for CRC and served as a novel biomarker for forecasting responses to immunotherapeutic interventions.

Purpose This study explored risk factors and gut microbiome characteristics associated with postoperative bowel functional recovery in patients undergoing colorectal surgery. Methods Patients undergoing colorectal surgery between January 2023 and October 2023 were stratified into two cohorts based on the timing of their first postoperative defecation (≤ 5 days vs. >5 days). Clinical data were systematically recorded to identify independent risk factors for delayed bowel recovery. Fresh postoperative stool specimens were collected and analyzed using metagenomic sequencing to examine the relationship between gut microbiota composition and bowel functional outcomes. Results Thirty-five patients were enrolled. Multivariate analysis identified the timing of first postoperative enteral feeding ( p < 0.01) as an independent risk factor for delayed defecation. Alpha diversity indices showed no significant differences between groups in microbial species richness. However, patients with delayed defecation (> 5 days) exhibited a lower relative abundance of probiotic taxa (e.g., Bifidobacterium, Lactobacillus) and an increased abundance of pathogenic bacteria compared to the ≤ 5-day cohort. Metagenomic profiling further demonstrated impaired microbial metabolic pathways in the delayed recovery group, including reduced carbohydrate metabolism (e.g., glycolysis/gluconeogenesis) and amino acid metabolism (e.g., selenocysteine and taurine biosynthesis). Conclusions Early postoperative resumption of enteral nutrition and probiotics may enhance bowel functional recovery. The observed reductions in microbial-driven gluconeogenesis/glycolysis, selenocysteine, and taurine synthesis suggest dysregulation of these metabolic pathways may compromise intestinal mucosal repair and homeostasis, contributing to delayed postoperative recovery.

Background Low anterior resection syndrome frequently complicates sphincter-preserving rectal cancer surgery. This study aimed to evaluate the capacity of pre-reversal high-resolution anorectal manometry and clinical factors to predict major low anterior resection syndrome at three months post-stoma reversal. Methods This prospective cohort study enrolled 46 patients with low rectal cancer undergoing protective stoma reversal. Prior to reversal, clinical data, colonoscopy (for diversion colitis), and high-resolution anorectal manometry parameters were assessed. Bowel function was evaluated using the low anterior resection syndrome score at one and three months. Multivariable logistic regression identified independent predictors of major low anterior resection syndrome at three months, followed by the construction and internal validation of a predictive nomogram. Results The incidence of major LARS decreased from 80.4% at one month to 47.8% at three months. Pre-reversal first sensation volume negatively correlated with 3-month low anterior resection syndrome scores (ρ = -0.32, p = 0.030). Multivariable analysis identified diversion colitis (adjusted OR = 6.95, 95% CI: 1.47 to 44.30, p = 0.023) and lower first sensation volume (adjusted OR = 0.90, 95% CI: 0.81 to 0.98, p = 0.031) as independent predictors. Causal mediation analysis confirmed these factors operated independently (p = 0.904). The composite predictive model demonstrated reliable discrimination (AUC = 0.736) and maintained stability (optimism-corrected C-index = 0.712) during internal validation. Conclusions Reduced first sensation volume and diversion colitis independently predict mid-term major low anterior resection syndrome. The developed pre-reversal nomogram facilitates individualized risk stratification, highlighting a potential window for targeted preemptive interventions.

Background Tumor-associated macrophages (TAMs) are critically involved in colorectal cancer (CRC) progression, yet their spatial and metabolic heterogeneity across primary and metastatic microenvironments remains poorly defined, limiting therapeutic development.Methods Integrated single-cell RNA sequencing was performed on 998,204 cells from a multicenter cohort encompassing primary CRC, adjacent normal tissue, liver and lymph node metastases, and peripheral blood. Computational analyses included uniform manifold approximation and projection (UMAP) clustering, pseudotime inference, RNA velocity, and CellChat. Findings were validated using multiplex immunohistochemistry, spatial transcriptomics, and in vivo preclinical models.Results This study identified two distinct TAM populations: SPP1+ and SEPP1+ TAMs. SPP1+TAMs, associated with angiogenesis, immune evasion, and liver metastasis, were linked to poorer prognosis. This population exhibited tissue-specific differentiation trajectories and comprised two metabolic subtypes: a glycolytic ATP5F1E+ subtype enriched in primary tumors and lymph nodes, and an oxidative phosphorylation-dominant MT-CO1+subtype specific to liver metastases. Spatially, MT-CO1+SPP1+TAMs localized to oxygen-rich invasive margins, while ATP5F1E+SPP1+TAMs resided in hypoxic tumor cores. Functionally, SPP1+TAMs drove CD8+ T-cell exhaustion and spatial exclusion. In preclinical models, combined regorafenib and anti-programmed cell death protein 1 therapy reduced SPP1+TAM infiltration and suppressed tumor growth and metastasis.Conclusion This study delineates the spatial, developmental, and metabolic heterogeneity of SPP1+TAMs in CRC, identifying SPP1+TAMs as key mediators of immunosuppression and metastasis. Targeting these populations with combination therapy effectively enhances antitumor immunity, presenting a novel translational strategy for CRC immunotherapy.

Tumor microenvironment (TME) serves as a critical mediator in cancer progression and treatment response, with cancer-associated fibroblasts (CAFs) recognized as core regulators of stromal-tumor interactions. As essential components of the TME, CAFs orchestrate tumor progression and therapy resistance through multifaceted mechanisms, including extracellular matrix (ECM) remodeling, angiogenesis modulation, and immune regulation, which collectively establish a complex regulatory network. Owing to their functional heterogeneity and potent tumor-promoting properties, CAFs have emerged as promising targets for novel anti-tumor therapies. This review highlights recent advances in the understanding of CAFs, focusing on their intricate interactions within the TME and their synergistic roles in promoting therapy resistance in cancer. Furthermore, we also discuss current CAF-targeted therapeutic strategies and ongoing clinical trials, and propose future research directions to further advance this field.

BACKGROUND:Stanniocalcin 1 (STC1) has been implicated in cancer pathogenesis, yet its pan-cancer implications and mechanistic roles in tumor progression and immune modulation remain incompletely characterized. The clinical relevance of STC1 in predicting prognosis and its interaction with tumor immune microenvironment components requires systematic investigation. OBJECTIVE:This study aims to establish the pan-cancer prognostic significance of STC1 and elucidate its associations with immunological characteristics, including immune checkpoint proteins, tumor mutational burden (TMB), microsatellite instability (MSI), and immune cell infiltration. This study focuses specifically on validating its role in the pathogenesis of gastric adenocarcinoma (STAD). METHODS:Multi-omics analysis was performed using TCGA pan-cancer datasets and bioinformatics tools (UALCAN, cBioPortal, HPA, GTA). Experimental validation included multiplex fluorescence staining of STAD tissue microarrays (n=30) and Western blot analysis of STAD cell lines. Key parameters analyzed encompassed clinical outcomes, cancer stemness indices, neoantigen load, and epithelial-mesenchymal transition (EMT) signatures. RESULTS:Pan-cancer analysis revealed significant STC1 overexpression in 18/33 cancer types (54.5%), particularly in prostate adenocarcinoma (94% deep deletions). STC1 expression correlated with poor prognosis (HR=1.32, p<0.01), elevated TMB (r=0.43), and MSI (r=0.38) across multiple malignancies. Single-cell RNA sequencing demonstrated a strong association with EMC (NES=2.18, FDR<0.001). In STAD, this study confirmed 3.7-fold protein overexpression (p=0.008) and identified positive correlations with CD8+ T cell infiltration (r=0.62, p=0.002) and CD4+ T cell infiltration (r=0.58, p=0.004). CONCLUSION:This multi-modal study establishes STC1 as a novel pan-oncogenic factor with dual roles in tumor progression (via EMT and stemness regulation) and immune microenvironment remodeling. The strong association with immune checkpoints (PD-L1, CTLA4) and T cell infiltration patterns positions STC1 as a promising immunotherapeutic target, particularly in STAD and MSI-high cancers. These findings provide mechanistic insights for developing STC1-directed therapeutic strategies.

e23514 Background: Gastrointestinal stromal tumor (GIST) is the most prevalent mesenchymal neoplasm of the gastrointestinal tract. While radical resection is the gold-standard treatment for localized GIST, the 5-year recurrence rate of high-risk GIST exceeds 50%, posing a major clinical challenge. Whether ctDNA-based detection of postoperative molecular residual disease (MRD) can reliably predict recurrence in high-risk locally advanced GIST patients remains a key question in translational oncology. Methods: This prospective study enrolled patients with high-risk locally advanced GIST who underwent R0 resection. Surgical tissue specimens were obtained for genomic profiling. Blood samples were collected at baseline (pre-surgery), at a landmark time point within one month after surgery, and serially thereafter at 3- to 6-month intervals during follow-up. Tumor-derived variants were identified through whole-exome sequencing of resected tumors. A personalized tumor-informed assay was designed by selecting up to 50 top-ranked variants with a variant allele frequency ≥3.0% to monitor MRD status. Results: As of Jan 2026 (cutoff), 44 eligible patients were enrolled. 44 tumor samples, 42 pre-surgical plasma samples, and 330 post-surgical blood samples were analyzed, with a median follow-up of 21 months. Tumor sequencing detected mutations in KIT and PDGFRA in 39 (88.6%) and 4 (9.1%) patients. Prior to surgery, MRD was detected in 57.1% (24/42) of plasma samples, and positivity correlated with larger tumor volume, higher Ki-67 index, mitotic count, and TMB (all P < 0.05). Multivariable logistic regression identified the Ki-67 index as an independent predictor of pre-surgical MRD positivity (OR, 1.20; 95% CI, 1.02 to 1.41; P = 0.024). In landmark analysis, ctDNA was detectable in 4 patients (4/41, 9.8%). These patients had worse DFS trend vs. MRD-negative patients (HR=3.48, 95% CI=0.70–17.35, P =0.11). To date, 8 patients had radiologically confirmed recurrence via CT. Among 4 patients with both MRD positivity and recurrence, one had longitudinal MRD positivity at 9 months post-op (3-month lead before CT relapse), and 2 had simultaneous detection by both methods. Notably, none of these 4 patients received regular adjuvant therapy. Additionally, multivariable Cox regression (adjusted for sex, age, TNM stage, and adjuvant therapy) showed longitudinal MRD positivity was associated with inferior DFS (HR=4.59, 95% CI=1.00–20.98, P =0.05). Patients with sustained negative MRD (n=29) had better survival than those with MRD conversion to negative (n=2), positive (n=8), or persistent positivity (n=2) (P=0.1). Conclusions: This study indicates that personalized tumor-informed ctDNA may predict recurrence in high-risk locally advanced GIST patients, particularly those not receiving regular adjuvant therapy. Clinical follow-up is ongoing. Clinical trial information: NCT05408897 .

The application of immunoscore (IS) in non-metastatic colorectal cancer has been validated. We performed a meta-analysis to evaluate the prognostic value of IS on survival in patients undergoing liver resection for colorectal liver metastasis (CRLM). Following the PRISMA 2020 guidelines, the protocol was registered in PROSPERO under number CRD42024498641. PubMed, Embase, Web of science, and the Cochrane Register of Controlled Trials databases were searched. To evaluate and compare the prognostic value of the IS, we independently assessed both the liver metastasis (metastatic IS) and the matched primary tumor (primary IS) for each patient with CRLM. Survival indicators include OS, RFS, DFS, with sufficient data to calculate hazard ratios (HRs) and 95

Microsatellite-stable/proficient mismatch repair (MSS/pMMR) colorectal cancer (CRC) is characterized by a cold tumor microenvironment, with limited CD8⁺ T cell infiltration and poor responsiveness to immune checkpoint inhibitors (ICIs). Here, using an in vivo CRISPR/Cas9 screen in a CMT93 cell-derived murine tumor model, we identify Arid3b as a key negative regulator of CD8⁺ T cell infiltration and antitumor activity. Genetic ablation of Arid3b in CD8⁺ T cells significantly enhances their intratumoral accumulation and promotes robust tumor control. Mechanistically, Arid3b deficiency upregulates Runx3, driving a tissue-resident memory-like phenotype and effector function. Notably, the benefits conferred by Arid3b deficiency are abrogated upon Runx3 deletion, indicating a RUNX3-dependent mechanism. Together, targeting ARID3B could offer a promising strategy to reshape the tumor microenvironment and sensitize MSS CRC to immunotherapy.

Abstract Background: Methylation-based analysis of cell-free DNA (cfDNA) has emerged as a key technology for MCED. However, existing approaches rely on traditional machine learning algorithms, which inherently limit detection performance. With the rapid advancement of artificial intelligence (AI), we have developed Genie-ADLA, a deep learning algorithm designed specifically for MCED. By integrating state-of-the-art deep neural network architectures with the intrinsic patterns inherent in methylation data, Genie-ADLA significantly enhanced MCED performance. Methods: Genie-ADLA was trained and evaluated on a dataset of 4,781 participants aged 40-75 years, including 2,702 pathologically confirmed cancer cases across 16 cancer types and 2,079 non-cancer controls (NCT06217900). The training set comprised 3,217 samples (1,756 cancer cases and 1,461 non-cancer controls), and the model’s performance was evaluated on an independent test set of 1,564 samples (618 non-cancer controls and 946 cancer cases). To address challenges inherent to methylation data—high dimensionality, sparsity, and noise—we applied feature dimensionality reduction and embedding strategies, reducing computational burden, mitigating overfitting, and improving learning efficiency. An ensemble learning approach further strengthened robustness and generalization. Results: Across all stages of 16 cancer types, Genie-ADLA achieved an overall sensitivity of 63.43% (600/946, 95% CI: [60.26%, 66.50%]) at 99.3% (612/618, 95% CI: [97.90%, 99.64%]) specificity in the test cohort. Compared with the XGBoost model trained on the same dataset, Genie-ADLA demonstrated improved overall sensitivity in 11 of the 16 cancer types, with an average increase of 4.86%.For stage I-III cancer patients, the sensitivities at 99.3% specificity showed notable gains over XGBoost: colorectal cancer achieved 76.98% (97/126, 95% CI: [68.65%, 84.01%]), an improvement of 9.52% from 67.46%; esophageal cancer reached 80.95% (51/63, 95% CI: [69.09%, 89.75%]), up 6.35% from 74.60%; breast cancer reached 37.14% (26/70, 95% CI: [25.89%, 49.52%]), improving by 5.71% from 31.43%. Lung cancer was subdivided into adenocarcinoma and non-adenocarcinoma, with stage I-III sensitivities of 40.90% (27/66, 95% CI: [28.95%, 53.71%]) in adenocarcinoma, an increase of 10.6%, and 84.44% (38/45, 95% CI: [70.54%, 93.51%]) in non-adenocarcinoma, improving by 2.22%. Conclusions: Genie-ADLA, leveraging advanced deep neural network architectures and data processing strategies, substantially elevates the performance ceiling of methylation-based early cancer detection, offering a new paradigm for AI-driven cancer screening. Citation Format: Kezhong Chen, Ziyu Li, Xiaojian Wu, Jian Huang, Guoyue Lv, Weiping Wen, Dahong Zhang, Xiangyu Zhao, Danbo Wang, Zhihua Liu, Lixin Sun, Shu Wang, Xiangnan Li, Zhigang Li, Jiandong Tai, Jiayin Yang, Zhentong Wei, Ming Cai, Qiang Zhang, Songbing He, Shuhua Yi, Shenhong Qu, Wenhui Zhao, Xianjun Yu, Ruixia Guo, Jianhong Lian, Desong Yang, Huaiwu Lu, Xi Guo, Yan Zhang, Zhuowei Liu, Yingjiang Ye, Chang Lin, Jie Gao, Xuanhui Liu, Yushu Guo, Suying Ding, Guoqiang Zhao, Yanzhan Yang, Jiangyu Li, Shiqing Chen, Hui Yu, Fang Liu, Yang Wang, Min Li, Baoliang Zhu, Yonghui Li, Xiaohui Wu, Fan Yang, Jun Wang. Genie-ADLA: A deep learning algorithm for methylation-based multiple cancer early detection (MCED) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2026; Part 1 (Regular Abstracts); 2026 Apr 17-22; San Diego, CA. Philadelphia (PA): AACR; Cancer Res 2026;86(7 Suppl):Abstract nr 5471.

Elevated intratumoral immune inflammation prior to treatment is typically associated with better outcomes in hot tumors treated with immune checkpoint inhibitors (ICIs). However, we observed a paradox in pMMR/MSS locally advanced rectal cancer (LARC) patients, where a subset with elevated baseline immune inflammation exhibited worse outcomes after combined radiotherapy and ICI treatment compared with patients with minimal immune inflammation. To investigate this counterintuitive finding, we performed paired scRNA-seq and scTCR-seq on longitudinally collected samples, including tumor biopsies (pre-treatment, post-radiotherapy, and post-immunotherapy) and peripheral blood mononuclear cells (pre-treatment and post-immunotherapy), from 20 pMMR/MSS LARC patients treated with sequential radiotherapy and ICI therapy (NCT06493240). We propose the concept of clonal entrapment to explain this phenomenon. Specifically, our profiling results reveal that increased HLA-DQA2 expression in dendritic cells and upregulated GDF15 expression in treatment-resistant tumor cells correlate with the restricted expansion of novel tumor-reactive TCR clonotypes. Consequently, the immune response is limited primarily by pre-existing TCR clonotypes within the tumor, especially those partially expanded under chronic inflammation, leading to the expansion of TCR clonotypes derived mainly from pre-treatment CD8+ T cell pools following ICI therapy. By identifying this feature of the pMMR/MSS LARC microenvironment, our study provides a high-resolution framework for understanding resistance to sequential radiotherapy and ICI therapy.

BACKGROUND:Perioperative chemo-immunotherapy shows variable outcomes in resectable gastric or gastro-oesophageal junction adenocarcinoma. We evaluated the efficacy and safety of neoadjuvant serplulimab with S-1 plus oxaliplatin (SOX) chemotherapy followed by adjuvant serplulimab versus neoadjuvant placebo plus SOX followed by adjuvant SOX in patients with PD-L1-positive, locally advanced, resectable gastric or gastro-oesophageal junction adenocarcinoma. METHODS:In this randomised, double-blind, multicentre phase 3 ASTRUM-006 study, patients were screened at 75 hospitals in China and Thailand. Eligible patients aged 18-70 years with PD-L1 combined positive score (CPS) ≥5, resectable gastric or gastro-oesophageal junction adenocarcinoma were randomly assigned 1:1 to neoadjuvant serplulimab or placebo (intravenous; 4·5 mg/kg) plus SOX chemotherapy (intravenous oxaliplatin 130 mg/m2 on day 1, and oral S1 40-60 mg twice daily on days 1-14) for three cycles (cycle length 21 days), followed by adjuvant serplulimab (up to 17 cycles; serplulimab group) or SOX (five cycles; placebo group). The primary endpoint was investigator-assessed event-free survival (defined as the time from randomisation to the occurrence of progressive disease or local or distant recurrence, other new malignancies, or death), with efficacy first evaluated in PD-L1 CPS ≥10, then in the intention-to-treat (CPS ≥5) population. This study is registered with ClinicalTrials.gov, NCT04139135, and is ongoing. FINDINGS:Between Nov 26, 2019, and April 19, 2024, 1646 patients were screened, of whom 588 patients (median age 61·0 years [IQR 55-66]; 124 [21%] female and 464 [79%] male) at 57 hospitals in China were randomly assigned to the serplulimab group (n=292) or placebo group (n=296). With a median follow-up duration of 42·7 months (IQR 24·3-53·6), median event-free survival was significantly longer with serplulimab than with placebo in the PD-L1 CPS ≥10 population (not reached [NR] vs 42·0 months; hazard ratio 0·65 [95% CI 0·47-0·90]; p=0·0082). With a median follow-up of 35·9 months (IQR 23·5-49·4), median event-free survival was also significantly longer with serplulimab than with placebo in the intention-to-treat population (NR vs 35·9 months; 0·73 [95% CI 0·56-0·94]; p=0·015). Grade 3 or worse treatment-related adverse events occurred in 136 (47%) patients in the serplulimab group and 172 (59%) patients in the placebo group; 19 (7%) and 31 (11%) patients in the respective groups discontinued treatment due to treatment-related adverse events. INTERPRETATION:Neoadjuvant serplulimab plus SOX followed by adjuvant serplulimab significantly improved event-free survival and demonstrated a better safety profile compared with neoadjuvant and adjuvant SOX in PD-L1-positive, resectable gastric or gastro-oesophageal junction adenocarcinoma. Extended follow-up for the overall survival data is warranted to confirm a survival advantage of this perioperative strategy with a chemotherapy-sparing adjuvant component for this indication. FUNDING:Shanghai Henlius Biotech.

Abstract The liver is the primary target organ for hematogenous metastasis of colorectal cancer, and colorectal cancer liver metastasis is one of the key and challenging aspects in its treatment. In order to improve the diagnosis and comprehensive treatment of colorectal cancer liver metastasis, the guideline development group has summarized advanced experiences and the latest achievements from both domestic and international sources, and has once again revised and updated the Guideline for the diagnosis and comprehensive treatment of colorectal cancer liver metastases (2025 edition) to continuously provide guidance and reference for clinical practice in this field.

Understanding the relationship between age and prognosis of female lymph node-metastatic gastric cancer (FLMGC) remains limited. This study aimed to elucidate the prognostic impact of age on FLMGC. This population-based retrospective analysis used data from the SEER database. After applying the exclusion criteria, a final cohort of 1,172 FLMGC cases was included. The patients were categorized by age into five groups: ≤ 50 years (n = 144), 51–60 years (n = 192), 61–70 years (n = 291), 71–80 years (n = 325), and ≥ 81 years (n = 220). Cancer-specific survival (CSS) was selected as the outcome in this study. Significant factors for CSS identified in multivariate analysis included age, tumor differentiation, AJCC T and N stages, systemic therapy, number of positive lymph nodes, and total number of retrieved lymph nodes. Age had a nonlinear effect on CSS (P for nonlinearity < 0.001), with the lowest risk occurring in the 55–60 year age group. This SEER-based study revealed a nonlinear relationship between age and FLMGC prognosis, with the lowest risk occurring in the 55–60-year age group.

Cell-free DNA (cfDNA) serves as a non-invasive biomarker for cancer detection, but conventional methods face challenges due to the ultra-low abundance of tumor-derived cfDNA (ctDNA) among normal cfDNA. Though nucleosome-bound cfDNA harbors rich epigenomic features that could enable ctDNA identification by single-molecule multi-omics cross-validation, this remains unexplored due to methodological limits. Here, we developed a cfDNA sequencing approach integrating methylation, fragmentomics, and histone modifications at the single-molecule level; together with gene semantics and epigenomic annotations, these modalities were vectorized and fused to represent each cfDNA molecule. We trained a Transformer-based model (cfAI) to profile and evaluate ctDNA likelihood at molecule, gene, and sample levels. cfAI achieved ~10-fold enrichment of cancer-derived signals over noise and reached 72.6% sensitivity at 93.1% specificity for multi-cancer detection. Our study establishes an innovative framework that overcomes the inherent signal-to-noise limitations of conventional assays and reveals biological features at molecular resolution for cancer detection. ### Competing Interest Statement Liyang Song, Ying Xin, Guoqiang Zhao, Guo Chen, Jing Liu, Baoliang Zhu, Xueguang Sun and Xiaohui Wu are employees of Shanghai Xiaohe Medical Laboratory Co., Ltd, Shanghai, China. Liyang Song, Xueguang Sun and Xiaohui Wu are authors of the related patents. All other authors have declared no conflicts of interest. Shanghai Xiaohe Medical Laboratory Co., Ltd.

Liver metastasis is a common complication in colorectal cancer (CRC), with its presence and progression significantly shortening patient survival. Therefore, a deeper understanding of the underlying mechanisms driving liver metastasis in CRC is essential to identify more effective and actionable therapeutic targets and improve prognosis. Liver metastasis in CRC is a multifaceted and dynamic process. Tumor cells with invasive properties communicate with the surrounding microenvironment through mechanisms such as immune checkpoint molecules and cytokines, thereby establishing a supportive niche for their colonization and proliferation. Moreover, suppressive immune cells may enhance the invasiveness of tumor cells. The interplay between tumor cells and the microenvironment is an interdependent process. Targeting these interactions offers promising potential for novel therapeutic strategies. The present review outlined mechanisms of colorectal cancer liver metastasis, emphasizing the immune microenvironment's role, current treatment approaches, and future development prospects.

Objective: Advanced gastric cancer remains highly refractory to therapy, with limited immunotherapy efficacy due to tumor microenvironment heterogeneity. Primary cilia, microtubule-based organelles involved in tumor progression, remain insufficiently explored in gastric cancer. This study aimed to define primary cilia subtypes and establish prognostic signatures for personalized treatment strategies.Methods: Bulk transcriptomic data from over 1,500 gastric cancer samples were integrated to define distinct primary cilia subtypes. A primary ciliary phenotype-associated signature（PCS） was established using a multimachine learning survival framework incorporating ten algorithms. The prognostic predictive value and immunotherapy response prediction capability of PCS were validated across multiple independent cohorts. Singlecell RNA sequencing analysis was performed to identify cellular populations associated with high-PCS phenotype.Causal weighted gene co-expression network analysis（WGCNA） was employed to identify driving factors, followed by functional validation through cell culture experiments and xenograft models.Results: Two distinct primary cilia subtypes were identified and validated across all cohorts, with C2 patients exhibiting significantly worse overall survival compared to C1 patients. PCS demonstrated robust predictive value for both prognosis and immunotherapy response, with superior accuracy compared to existing models across multiple validation cohorts. High-PCS patients showed reduced tumor purity, increased stromal cell infiltration,and poor response to immunotherapy. Single-cell analysis revealed that fibroblasts had the highest PCS scores and identified a novel secreted modular calcium-binding protein 2（SMOC2） high myofibroblastic cancer-associated fibroblast（mCAF） population as the key driver of high-PCS phenotype. Functional experiments confirmed that SMOC2 knockdown significantly suppressed gastric cancer cell proliferation, migration, and invasion, while promoting mCAF-to-inflammatory cancer-associated fibroblasts（iCAF） transition.Conclusions: PCS serves as a robust prognostic biomarker for gastric cancer patients. Additionally, targeting SMOC2 high mCAFs represents a potential therapeutic strategy for patients with high-PCS gastric cancer.

BackgroundRecent years, immunotherapy has emerged as a pivotal approach in cancer treatment. However, the response of gastric cancer to immunotherapy exhibits significant heterogeneity. Therefore, the early identification of gastric cancer patients who are likely to benefit from immunotherapy and the discovery of novel therapeutic targets are of critical importance.Materials and methodsWe collected data from European Nucleotide Archive (ENA) and Gene Expression Omnibus (GEO) databases. In project PRJEB25780, we performed WGCNA analysis and Lasso regression and chose CXCR2P1 for the subsequent analysis. Then, we compared the expression difference of CXCR2P1 among different groups. Kaplan-Meier curve was used to analyze the prognostic value of CXCR2P1, which was validated by project IMvigor210 and GEO datasets. ESTIMATE and CIBERSORT algorithm were used to evaluate the reshaping effect of CXCR2P1 to immune microenvironment of tumor. Differentially expressed genes (DEG) analysis, enrichGO analysis, Gene Set Enrichment Analysis (GSEA) and co-expression analysis were used to explore the cell biological function and signaling pathway involved in CXCR2P1.ResultsWGCNA identified CXCR2P1 as a hub gene significantly associated with immune response to PD-1 inhibitors in gastric cancer. CXCR2P1 expression was elevated in responders and correlated with better prognosis. Functional analysis revealed its role in reshaping the tumor immune microenvironment by promoting immune cell infiltration, including M1 macrophages, activated CD4+ T cells, and follicular helper T cells. CXCR2P1 enhanced antigen presentation via the MHC-II complex, influenced key immune pathways, such as Toll-like receptor signaling and T-cell activation, which led to the up-regulation of expression of PD-L1. GSEA showed CXCR2P1 were correlated with microRNAs. Through DEG analysis and expression analysis, MIR215 was identified as a potential direct target of CXCR2P1.ConclusionHigh expression of CXCR2P1 is correlated with better response to PD-1 inhibitor. It reshapes the immune microenvironment by increasing immune infiltration and changing the fraction of immune cells. In tumor immune microenvironment, CXCR2P1 can promote inflammation, enhance antigen presentation and activate the PD-1/PD-L1-related signaling pathway, which might be achieved by CXCR2P1-MIR215 axis.