222 Background: Gestational trophoblastic neoplasia (GTN) is caused by cancerous transformation of placental trophoblastic cells, which can undergo hematogenous spread via circulating tumor cells (CTCs). Due to limited accessibility to tumor tissues, the molecular classification and metastatic evolutionary dynamics of GTN progression remains poorly defined. Methods: We performed molecular subtyping and evolutionary trajectory analysis using single-cell omic profiling of CTCs and patient-matched tumor cells from GTN blood and tissue biopsies respectively. Experimental in vitro and in vivo assays were employed to test the function of disease-driving genes involved in GTN progression. Blood and tissue-based clinical analyses from independent patient samples were carried out to demonstrate the molecular heterogeneity of GTN subclones within individual patients. Results: Three distinct GTN molecular subtypes were identified and validated in clinical samples, including cilia-like subtype marked by FOXJ1 positivity, tuft cell-like subtype driven by POU2F3, and a senescent subset with elevated BHLHE41 expression. Experimental analyses using GTN cell line models confirmed a central role of POU2F3 in driving epithelial–mesenchymal transition (EMT), migration, invasion, and tumorigenesis in vivo . Furthermore, BHLHE41 overexpression is sufficient to drive p53/p21 activation and senescence induction, characterized by β-Galactosidase formation and elevated expression of the senescence-associated secretory phenotype (SASP) signatures. In a prospective cohort of patient blood samples, CTCs overexpressing POU2F3 or BHLHE41 were significantly elevated in the GTN group compared to the benign hydatidiform moles (HM) cases, implicating their translational potential as single-cell liquid biopsy for accurate molecular subtyping and prediction of GTN progression in the absence of tissue biopsies. Conclusions: This study uncovers key molecular subtypes and functional states of GTN using CTC-based single-cell omic analysis, offering a noninvasive approach for disease classification and real-time monitoring in the absence of tumor tissue biopsies.

The precise activation of the cGAS-STING pathway for effective tumor immunotherapy remains a significant challenge due to the complexity of immune responses and tumor microenvironment (TME) limitations. Here, a multifunctional nanoagonist, cRGD-PDA/ZIF8@ICG/TPT (cDZ@IP), was developed to realize nano-metabolite-driven multimodal synergistic activation of the STING pathway and enhanced immune recognition. The agonist combines cRGD peptide-targeted polydopamine-coated zeolitic imidazolate framework-8 with the photosensitizer indocyanine green and the chemotherapeutic drug topotecan. Upon near-infrared laser irradiation, cDZ@IP degrades within the TME, generating high levels of reactive oxygen species, inducing mitochondrial stress, and releasing endogenous mitochondrial DNA. Additionally, topotecan enhances DNA damage accumulation by inhibiting nuclear DNA repair. Zn2 + released from the agonist further amplifies cGAS-STING pathway activation, thereby ensuring a robust immune response. The mild photothermal therapy-induced immunogenic cell death promotes the initiation of antitumor immunity, while also enhancing the effectiveness of immune checkpoint blockade. In vivo studies show that cDZ@IP significantly inhibits primary and distant tumor growth and prevents lung metastasis, providing a promising strategy for STING pathway-targeted cancer immunotherapy.

LBA3515 Background: Total neoadjuvant chemoradiotherapy is the standard of care for locally advanced rectal cancer (LARC) to control local recurrence and achieve organ preservation. However, for proficient mismatch repair (pMMR) or microsatellite stable (MSS) LARC, which accounts for nearly 90% of rectal cancers, conventional chemoradiotherapy has limited efficacy and is associated with significant side effects. Recent studies have shown that combining radiotherapy with immunochemotherapy can improve pathological complete response (pCR) rates, but the inclusion of tumor-draining lymph nodes (TDLNs) in the conventional irradiation field may impair T-cell immunity and reduce response to immunotherapy. Our previous single-arm phase II trial demonstrated that node-sparing modified short-course radiotherapy combined with chemotherapy and PD-1 blockade could achieve a high pCR rate of 78.8% in pMMR LARC. 1 Building on these findings, we initiated this phase III trial to compare this new treatment regime with conventional short-course chemoradiotherapy in improving pCR rates. 2 Methods: This is a phase III, open-label, multicenter, randomized trial conducted across 17 hospitals in China. A total of 154 eligible MSS/pMMR middle or low rectal cancer patients (cT3-4N0/+M0) will be recruited and randomly assigned (1:1) to two groups: control group (conventional short-course chemoradiotherapy), experimental group (node-sparing modified short-course chemoradiotherapy plus PD-1 blockade). The innovative node-sparing modified short-course radiotherapy targets only the primary tumor bed, excluding TDLNs. Following randomization, patients will receive short-course radiotherapy (conventional or node-sparing) followed by four cycles of CAPOX ± tislelizumab: tislelizumab 200 mg IV on day 1, oxaliplatin 130 mg/m² IV on day 1, and capecitabine 1000 mg/m² orally on days 1-14, and Total mesorectal excision (TME) will be performed at weeks 14-15. The primary endpoint is pCR rate, while secondary endpoints include organ preservation rate, disease-free survival, overall survival, adverse effects, and quality of life. The primary and secondary endpoints will be analyzed in the intent-to-treat (ITT) population. Safety analyses will be performed in the safety population, defined as patients who received at least one dose of study treatment. Results: 247 patients were assessed for eligibility, a total of 154 patients were enrolled in the ITT population (77 per group). Baseline characteristics were well balanced between the two groups. In the ITT population, the experimental group demonstrated a significantly superior pCR rate compared to the Control group: 61.4% (47/77) vs 28.6% (22/77) (P < 0.001). The MPR rate was also significantly higher in the experimental group (80.5% vs 50.6%). All patients in both groups received sphincter-sparing surgery. Regarding safety, the incidence of grade 3-4 treatment-related adverse events (TRAEs) was comparable between the experimental and control groups (23.4% vs 22.1%). Immune-related adverse events (irAEs) occurred in 7.8% (6/77) of patients in the experimental group, primarily grade 1-2. Conclusions: Compared with conventional short-course preoperative chemoradiotherapy, node-sparing modified radiotherapy combined with CAPOX and PD-1 blockade significantly improved pCR rates in patients with pMMR/MSS LARC, with a manageable safety profile. This regimen represents a promising and highly effective neoadjuvant strategy for MSS rectal cancer. Clinical trial information: NCT06507371. Reference：1. Annals of Oncology (2024) 24 (suppl_1): 1-20. 10.1016/iotech/iotech100744; 2. 2025;43 16_suppl. https://doi.org/10.1200/jco.2025.43.16_suppl.tps3641. Clinical trial information: NCT06507371 .

Emerging evidence suggests that tissue-resident microbiota (TRM) is associated with tumor biology; however, their distribution and compositional characteristics across different colorectal tissue types remain incompletely defined. Here, we conducted a comprehensive cross-sectional analysis of TRM distribution and abundance across 1134 clinical specimens, including normal mucosa, precancerous polyps, and colorectal cancer (CRC) tissues. Our results reveal distinct microbial profiles among these diagnostic groups, with consistent differences in community composition between normal, polyps, and CRC samples. Integrative analyses further identified microbial signatures capable of distinguishing tissue categories and reflected differences in the local tumor microenvironment. In contrast, intratumoral microbiota composition showed subtle variation across established tumor stages and was not associated with clinical outcomes. These findings define diagnostic group-associated patterns of TRM in colorectal tissues and establish a foundation for future mechanistic investigations into the biological roles of TRM in colorectal cancer.

Immunotherapy has reshaped the treatment landscape of colorectal cancer (CRC), with the clearest and most durable benefit established in mismatch repair-deficient (dMMR)/microsatellite instability-high (MSI-H) disease. However, framing CRC immunotherapy simply as "MSI-H responsive versus microsatellite stable (MSS) resistant" is no longer sufficient. Recent studies indicate that a subset of proficient mismatch repair (pMMR) colon cancers, particularly in the neoadjuvant setting, can mount clinically meaningful responses to immune checkpoint blockade, suggesting that disease stage, local immune organization, and treatment timing critically influence immunotherapy sensitivity. In parallel, emerging evidence has expanded the relevant immune landscape beyond the tumor bed itself, showing that spatially organized stromal and adipose niches can actively divert tumor-reactive lymphocytes and promote immune escape. These advances shift the central challenge in CRC immunotherapy from simply identifying new agents to defining when and in whom immune resistance is reversible, and which biological bottlenecks-such as vascular dysfunction, myeloid suppression, and spatial immune exclusion-must be overcome. In this context, alternative checkpoint inhibitors, bispecific antibodies, cellular therapies, vaccines, nanotechnology-enabled platforms, and microbiome-targeted approaches remain important, but their translational maturity and evidentiary support differ substantially. Biomarker development is likewise evolving from static genomic classification toward dynamic and mechanism-informed stratification incorporating circulating tumor DNA (ctDNA), chromosomal instability, immune architecture, and treatment-induced response trajectories. This review synthesizes recent advances in CRC immunotherapy while emphasizing evidence hierarchy, biomarker-guided patient selection, and the mechanistic basis of combination strategies. We argue that the next phase of CRC immunotherapy will depend less on the indiscriminate addition of novel agents and more on the rational deployment of immunotherapy across molecularly, spatially, and temporally defined disease states.

In the past 5 years, clinical trials on immune checkpoint inhibitors (ICIs) for the treatment of locally advanced rectal cancer (LARC) have flourished globally, and China has become one of the leading regions in this field. In response to the breakthrough progress and accumulation of evidence from key clinical trials, the Chinese Society of Colorectal Surgery has recognised the need for updated consensus guidance on the development of perioperative and organ-preserving treatment strategies for LARC. This expert consensus guidance provided unified standards for the indications, medication regimens, efficacy evaluations and follow-up of ICIs in this population, with a focus mainly on perioperative management and organ-sparing strategies. The diagnostic part of this consensus guidance is based on the internationally recognised definition of mismatch repair/microsatellite instability detection and emphasises the importance of multidisciplinary teams in treatment decision-making. In terms of treatment, based on the results of key trials that have changed clinical practice in the past 5 years, this expert consensus provides graded recommendations for the duration of preoperative immunotherapy and the necessity of postoperative adjuvant therapy, local resection and organ preservation strategies. Moreover, we refined the management process for the safety of perioperative immunotherapy. This document aims to provide a reference for surgeons; internal medicine, radiation therapy, pathology and imaging physicians; patients and nursing staff involved in the treatment of LARC, as well as health policy makers.

LBA3563 Background: The optimal second-line treatment for metastatic colorectal cancer (mCRC) after failure of oxaliplatin-based chemotherapy therapy remains an area of active research, while bevacizumab combined with chemotherapy is a standard option. Fruquintinib (Fru) is a highly selective and potent oral tyrosine kinase inhibitor of VEGFR 1, 2, and 3. This study aimed to compare the efficacy and safety of fruquintinib combined with chemotherapy versus bevacizumab combined with chemotherapy as second-line treatment for mCRC. Methods: This was a non-inferiority trial conducted across 12 hospitals and cancer centers in China. Patients with mCRC who had progressed on fluoropyrimidine and oxaliplatin-based first-line chemotherapy therapy were randomly assigned (1:1) to receive either Fru (4 mg orally, once daily for 3 weeks on/1 week off) plus FOLFIRI, or bevacizumab (Bev) (5 mg/kg intravenous, every 2 weeks) plus FOLFIRI. Randomization was stratified by primary tumor location and RAS/BRAF status. During combination therapy, pts who achieve disease control after 4-6 months of treatment proceeded to maintenance therapy, receiving either fruquintinib combined with capecitabine or bevacizumab combined with capecitabine. The primary endpoint was progression-free survival (PFS). Final analysis occurred after either 97 PFS events or after the last patient had completed 12 months of follow-up, whichever occurred first. The non-inferiority upper margin of HR was 1.33. Results: Between Jul 13, 2023, and Mar 4, 2025, 122 pts were enrolled and randomly assigned to the Fru group (n=60) or the Bev group (n=62). The median age was 59.0 (IQR 54-69) and 60.0 (IQR 51-70) years, 45 (75%) and 48 (77%) had left-sided tumors, 36 (60%) and 36 (58%) were RAS/BRAF mutant, respectively. At data cut-off (Feb 28, 2026), the median PFS of the Fru group was non-inferior to the Bev group, whether in the Intention to Treat Population (ITT) (9.40 vs 7.39 months, hazard ratio [HR]=0.806 [95% CI 0.522–1.244, 80% CI 0.607-1.07]; p=0.33) or the per-protocol set (PPS) (9.49 vs 7.85 months, HR=0.848 [95% CI 0.542–1.326, 80% CI 0.633-1.136]; p=0.469). In the subgroup analysis , the Fru group showed longer PFS in prior never used VEGF inhibitor pts (10.6 vs 8.5 months, HR=0.94, 95% CI 0.54-1.62), and pts without liver metastasis (14.5 vs 9.9 months, HR=0.97, 95% CI 0.43-2.22) compared to the Bev group. The objective response rate was 35.0% with Fru vs 22.6% with Bev group. Any-grade treatment-emergent adverse events (TEAEs) occurred in 100.0% (Grade ≥3, 28.1%) of the Fru group and 93.2% (Grade ≥3, 28.8%) of the Bev group. Conclusions: Fruquintinib combined with FOLFIRI demonstrated non-inferiority in PFS compared with bevacizumab combined with FOLFIRI as a second-line treatment for mCRC, with a manageable safety profile. Clinical trial information: NCT05555901 .

[This corrects the article DOI: 10.1016/j.isci.2023.106521.].

Full-thickness skin wounds remain a global clinical burden, perpetuated by impaired inflammation resolution, angiogenesis, and extracellular matrix (ECM) remodeling. Here, we report a sunlight-responsive hydrogel (A@HN: Au/g-C₃N₄@HAMA-NB) that synergizes ECM-mimetic scaffolding with photoelectric conversion to address these pathological barriers-realizing the vision of “promoting skin regeneration through sunlight exposure”. In both normal and diabetic Sprague-Dawley rats with full-thickness skin defects, the A@HN hydrogel combined with daily sunlight irradiation (A@HN (light)) achieved rapid wound closure, outperforming control groups by a substantial margin. Histologically, A@HN (light) promoted complete re-epithelialization, thick granulation tissue formation, and native-like collagen deposition, with tissue architecture approaching that of normal skin. Immunologically, it shifted macrophages toward a reparative M2 phenotype and enhanced angiogenesis, while transcriptomic analysis revealed activation of angiogenesis, ECM organization, and immune-modulatory pathways. These changes were accompanied by upregulated pro-regenerative factors and downregulated pro-inflammatory cytokines. Mechanistically, sunlight-triggered electrical signals orchestrate a multi-stage regenerative response: resolving inflammation, driving neovascularization, and remodeling ECM. This sunlight-responsive system offers a wireless strategy for general full-thickness skin defects and refractory diabetic wounds, with significant clinical translation potential.

Background: Accurate preoperative prediction of lymph node metastasis (LNM) is essential for tailoring treatment strategies in T1 colorectal cancer (CRC). Although the neutrophil-to-lymphocyte ratio (NLR), an easily acquired inflammatory biomarker, correlates with tumor progression, its incremental value when incorporated into machine learning (ML) models for predicting LNM in T1 CRC remains unclear. Therefore, this study evaluated the independent predictive value of preoperative NLR for LNM and constructed interpretative ML models to stratify LNM risk in T1 CRC. Methods: We retrospectively enrolled 533 pathologically confirmed T1 CRC patients. NLR was calculated as the neutrophil-lymphocyte count ratio. Multivariable logistic regression identified NLR's independent correlation with LNM. Features were screened via combined least absolute shrinkage and selection operator (LASSO) regression and the Boruta algorithm. Six ML models, including Logistic Regression (LR), Random Forest (RF), XGBoost, LightGBM, CatBoost, and Logistic Regression with Splines (LR-Spline), were developed and validated. Model performance was evaluated in terms of discrimination, calibration, and clinical utility. Shapley additive explanations (SHAP) were applied for feature interpretability. Results: Elevated preoperative NLR was independently associated with an increased risk of LNM (adjusted OR = 2.83, 95% CI: 1.48-5.41, p = 0.002), exhibiting stability across all clinical and pathological subgroups. The optimized LightGBM outperformed other models with an area under the receiver operating characteristic curve (AUC) of 0.768, alongside favorable clinical utility. Conclusions: An elevated preoperative NLR is a robust, independent predictor of LNM in patients with T1 CRC. An optimized LightGBM model integrating NLR with routine clinicopathological indicators offers accurate risk stratification, potentially refining surgical decision-making and sparing low-risk T1 CRC patients from unnecessary radical resections.