BackgroundUltra-radical cytoreductive surgery is frequently performed for patients with advanced ovarian cancer (OC). However, anastomotic leakage (AL) is a serious complication of such surgeries and the risk factors remain unclear. This study identified early postoperative C-reactive protein (CRP) and albumin were the strongest predictors of leakage.MethodsThis multicenter retrospective study involved 305 patients with ovarian cancer who underwent primary anastomosis following enterectomy, spanning January 2018 to June 2023. Comprehensive clinical and demographic data were used to develop predictive models. Feature selection was performed using LASSO and univariate logistic regression. Machine learning algorithms were subsequently applied, with model interpretability assessed using SHapley Additive explanations (SHAP).ResultsThe study revealed an AL prevalence of 14.1%, with 46.5% of affected patients requiring reoperation. Five predictors were identified, including postoperative CRP, serum albumin levels, Eastern Cooperative Oncology Group score, N stage, and blood urea nitrogen. The Lasso-Logistic model demonstrated the best predictive performance with an area under the curve of 0.828 (0.119). SHAP analysis highlighted early postoperative CRP, albumin, and N stage as major contributing factors. Economic analysis revealed a significant correlation between AL and hospital stay, hospital costs, and time to chemotherapy.DiscussionEarly postoperative inflammatory and nutritional biomarkers, particularly CRP and albumin, demonstrated significant predictive value for anastomotic leakage, providing an early warning for risk stratification and intervention. The investigation also bolstered the evidence supporting the restrictive surgical scope approach advocated in clinical guidelines.

ABSTRACT High‐grade serous ovarian cancer (HGSOC) is the most common and lethal histological subtype of ovarian cancer. This study aimed to identify peritoneal metastasis‐associated biomarkers of HGSOC to provide directions for subsequent treatment. Transcriptome data were extracted from the GEO database and literature. Differential expression analysis, a PPI network, and machine learning were conducted to obtain key cells and biomarkers, followed by the construction of ROC curves and expression validation to screen biomarkers. The functional mechanism of the biomarkers was explored via a molecular regulatory network and enrichment analysis. Potential drugs targeting biomarkers were also predicted. Based on the single‐cell and spatial transcriptome data, the expression patterns and communication types of key cells and biomarkers were explored. CYTH1 and ARHGEF1 were identified as biomarkers, while ILCs, DC‐1, and macrophages were identified as key cells associated with metastasis in HGSOC. Spatial transcriptome data confirmed that the biomarkers are highly expressed in T and NK cells. Cell interaction analysis indicated that the expression of SPP1 was higher in peritoneal metastases than in primary lesions. Furthermore, in vitro studies demonstrated that macrophage‐derived SPP1 significantly enhanced the proliferation and migration of ovarian cancer cells. This study identifies CYTH1 and ARHGEF1 as metastatic biomarkers and implicates macrophages in promoting metastasis through SPP1, which provides new insights into the understanding of tumor metastatic mechanisms in HGSOC.

e17511 Background: Circulating tumor HPV DNA (ctHPV) is emerging as a promising biomarker for minimal residual disease (MRD) in cervical cancer. Prospective studies investigating the association of ctHPV at diagnosis with primary tumor size and lymph node metastasis (LNM), as well as its potential for longitudinal MRD monitoring, are limited. Establishing a rapid and clinically feasible method for ctHPV quantification could improve early risk stratification and guide precision management strategies. Methods: Newly diagnosed HPV16/18-positive cervical cancer patients are being prospectively enrolled, with a planned cohort of 83 patients. 76 patients have been analyzed to date. ctHPV levels at diagnosis were quantified using droplet digital PCR (ddPCR) and next-generation sequencing (NGS). The concordance between ddPCR and NGS was evaluated using correlation analysis and Bland–Altman plots. Associations of ctHPV with LNM status and trends according to primary tumor size were analyzed. Post-treatment ctHPV measurements are ongoing, and patients will continue longitudinal follow-up to evaluate ctHPV dynamics and their potential association with treatment response and prognosis. Results: Analyses presented here include 76 patients from the ongoing prospective cohort. ctHPV was detectable at diagnosis in all patients. ddPCR showed high concordance with NGS, with Bland–Altman analysis demonstrating minimal bias, supporting its use as a rapid and clinically feasible quantification method. Patients with LNM exhibited higher ctHPV levels at diagnosis compared with those without LNM, and ctHPV levels increased with greater primary tumor size. These findings highlight the potential of ctHPV to reflect tumor burden and LNM status at diagnosis. Conclusions: ctHPV at diagnosis is associated with primary tumor size and lymph node metastasis in cervical cancer. ddPCR provides a rapid and reliable method for ctHPV quantification. Ongoing and future longitudinal analyses will explore ctHPV dynamics during treatment and follow-up as a marker for MRD and prognosis, supporting its potential clinical utility in prospective patient management. Circulating tumor HPV DNA (ctHPV) levels measured by ddPCR and NGS in the prospective cohort, showing stratified by lymph node metastasis and primary tumor size. Analysis/cohort N ddPCR log10 (ctHPV + 1) (copies/ml) NGS log10 (ctHPV + 1) (copies/ml) Mean difference (95% LoA) P Value (ddPCR) P Value (NGS) ddPCR/NGS concordance 76 2.091 ± 1.19 2.27 ± 1.29 -0.18 (-0.72 – 0.36) # - - LNM positive 37 2.58 ± 1.24 2.80 ± 1.41 - 0.0003 0.0003 LNM negative 39 1.63 ± 0.94 1.77 ± 0.93 - Tumor <= 2 cm 9 1.25 ± 0.91 1.46 ± 0.89 - 0.0002 * 0.0005 * Tumor 2 – 4 cm 21 1.57 ± 0.86 1.71 ± 0.79 - Tumor >= 4 cm 46 2.49 ± 1.20 2.69 ± 1.38 - #: Bland–Altman analysis; *: Linear trend was evaluated by polynomial contrasts in one-way ANOVA.

5544 Background: Persistent infection with high-risk human papillomavirus (HPV), particularly HPV16 and HPV18, drives the development of high-grade squamous intraepithelial lesions (HSIL), an obligate precancerous condition. The standard care of cervical HSIL requires either surgical excision (LEEP or cold-knife conization), or in some circumstance, watchful waiting. AFN0328 is a novel mRNA-based immunotherapy encoding HPV16/18-related antigens, designed to elicit antigen-specific immune responses and interrupt viral antigen-driven disease progression. Methods: Safety, tolerability and preliminary efficacy of AFN0328 were assessed in patients with HPV16- and/or HPV18-associated HSIL (CIN2/3) in an ongoing, multicenter, open-label Phase 1 trial. The study comprised two sequential parts: Part 1 employed a classic 3+3 dose-escalation design to assess safety and determine the maximum tolerated dose (MTD); Part 2 is a dose-expansion cohort at selected doses (150 μg and 300 μg) to further characterize safety, tolerability, and preliminary efficacy. AFN0328 was administered intramuscularly at 0, 4, and 12 weeks. The primary endpoints included safety and tolerability (dose-limiting toxicities within 28 days post-first dose, adverse events graded per CTCAE v5.0). For Part 2, the efficacy endpoints included viral clearance rate at Week 24 and Week 36, and histopathological regression at Week 36. Secondary endpoints included pharmacokinetic parameters, HPV16/18-specific IgG antibody titers, and pharmacodynamic markers (cellular immune responses and serum cytokines). Results: As of December 29, 2025, 23 patients had received at least one dose of AFN0328. No DLTs, serious adverse events, or grade ≥3 treatment-related adverse events (TRAE) were observed. PK analyses of first and last doses showed that circulating mRNA encoding HPV-related antigens peaked within 48 hours and declined gradually, with detectable levels persisting for up to at least 4 weeks across dose levels. All evaluable patients developed specific IgG responses and viral clearance was observed in evaluable patients during follow-up. Conclusions: AFN0328 demonstrated favorable safety and tolerability with robust immune responses in HPV16/18-associated HSIL, supporting further clinical development as an immunotherapy for HPV-driven precancerous disease. Clinical trial information: CTR20244013. Summary of safety and immunogenicity outcomes. Outcome Result Patients treated 23 Dose-limiting toxicities 0 Grade ≥3 treatment-related AEs 0 Most common TRAEs Injection-site reactions, fever Specific IgG response rate 100%* Observed viral clearance Yes* *Observed in evaluable patients during interim follow-up.

Metabolic reprogramming and immune evasion are central hallmarks of cancer progression; yet, how mitochondrial metabolism shapes immune escape remains unclear. Here, we identify GBP6 as a tumor-intrinsic mitochondrial factor associated with aggressive cervical cancer. GBP6 is enriched in high-risk tumors and correlates with poor clinical outcomes. GBP6 knockdown suppresses tumor cell proliferation and promotes cell death. Mechanistically, GBP6 localizes to mitochondria, interacts with TACO1, and supports respiratory complex IV integrity and activity. GBP6 depletion disrupts mitochondrial bioenergetics, reduces membrane potential and NADPH availability, increases ROS production, and compromises the GSH-GPX4 antioxidant axis, thereby promoting lipid peroxidation and ferroptosis-associated cell death. This mitochondrial redox stress induces ICAM-1 expression through ROS-NF-κB signaling, enhancing NK-cell adhesion, immune synapse formation, and tumor-cell killing. In xenograft models, GBP6 depletion sensitizes tumors to adoptively transferred NK cells. These findings identify GBP6 as a mitochondrial regulator linking metabolic fitness to innate immune evasion in cervical cancer.

Concurrent chemoradiotherapy (CCRT) followed by intrauterine brachytherapy is the standard treatment for locally advanced cervical cancer (LACC), yet its efficacy is frequently constrained by an immunosuppressive tumor microenvironment (TME). This randomized, single-center, Phase II trial (n = 18; 2:1 allocation; high-risk Stages III-IVA LACC) investigated whether adding immunotherapy to CCRT (CICRT) could enhance early tumor regression and overcome immune suppression. In this exploratory analysis, CICRT demonstrated a numerical advantage in tumor reduction over CCRT alone, although the difference did not reach conventional statistical significance (mean residual tumor volume: 3.0% vs. 7.4%, p = 0.080, 95% CI: -9.3%-0.6%, η 2 = 0.180). Single-cell RNA sequencing of paired biopsies (n = 12) revealed that CICRT was associated with pro-inflammatory remodeling, characterized by the upregulation of MHC-II and chemotaxis-related genes in malignant cells and cancer-associated fibroblasts. Notably, CICRT reduced regulatory T cell niches and exhausted CD8+ T cells, alongside a phenotypic shift in tumor-associated macrophages toward an inflammatory state. These findings demonstrate the potential of CICRT to enhance early tumor regression and mitigate TME suppression, providing an exploratory biological rationale for its therapeutic activity in LACC.

Taxane-based chemotherapy is a main treatment modality for ovarian cancer and other solid tumors, but chemoresistance limits the clinical efficacy. Studies have shown tumor interaction with macrophages in the tumor microenvironment (TME) plays a significant role in taxane resistance, yet the underlying molecular mechanisms are poorly understood. In this study, we employed translatome profiling of paclitaxel-treated cancer cells, live-cell imaging analysis, gene knockdown/knockout, and in vitro cancer-macrophage coculture assays to unravel a novel chemoresistance mechanism mediated by tumor-macrophage interaction via the NOTCH2-JAG1 axis. The in vitro data were further validated by multiple xenograft, syngeneic and patient-derived xenograft mouse tumor models of ovarian cancer as well as ovarian cancer patient samples. We found paclitaxel selectively induced translational upregulation of NOTCH2 via cytoplasmic polyadenylation, and this NOTCH2 upregulation persisted after mitotic exit. Subsequent NOTCH2 activation by JAG1 expressed mainly on the neighboring macrophages promoted tumor cell survival and simulated cytokine release, such as CSF1 and IL-1β, that recruited JAG1-expressing macrophages, thus forming a positive feedback loop that further enhanced the pro-tumor NOTCH2 activity. Genetic depletion or pharmacological inhibition of NOTCH2 with the γ-secretase inhibitor attenuated macrophage infiltration and sensitized tumor response to paclitaxel in multiple preclinical models of ovarian cancer. Moreover, single-cell RNA sequencing analysis identified a JAG1-high macrophage subset that was enriched by paclitaxel treatment and attenuated by NOTCH inhibition. Clinically, high NOTCH2 expression in ovarian tumors was associated with recurrence and shorter progression-free survival of ovarian cancer patients. Paclitaxel-induced translational upregulation of NOTCH2 enables immediate juxtacrine activation by JAG1-positive macrophages, coupling tumor cell survival with immune remodeling in the tumor microenvironment to drive chemoresistance. Our results suggest NOTCH2 is a viable biomarker for paclitaxel resistance and that combining NOTCH2 inhibitor with taxane is an effective therapeutic strategy to selectively disrupt tumor-macrophage interaction and overcome macrophage-mediated taxane resistance in NOTCH2-positive tumors.

Heat shock protein 90 (HSP90) is overexpressed in numerous cancers, promotes the maturation of numerous oncoproteins and facilitates cancer cell growth. Certain HSP90 inhibitors have entered clinical trials. Although less than satisfactory clinical effects or insurmountable toxicity have compelled these trials to be terminated or postponed, these results of preclinical and clinical studies demonstrated that the prospects of targeting therapeutic strategies involving HSP90 inhibitors deserve enough attention. Nanoparticulate-based drug delivery systems have been generally supposed as one of the most promising formulations especially for targeting strategies. However, so far, no active targeting nano-formulations have succeeded in clinical translation, mainly due to complicated preparation, complex formulations leading to difficult industrialization, incomplete biocompatibility or nontoxicity. In this study, HSP90 and CD44-targeted A6 peptide functionalized biomimetic nanoparticles (A6-NP) was designed and various degrees of A6-modification on nanoparticles were fabricated to evaluate targeting ability and anticancer efficiency. With no excipients, the hydrophobic HSP90 inhibitor G2111 and A6-conjugated human serum albumin could self-assemble into nanoparticles with a uniform particle size of approximately 200 nm, easy fabrication, well biocompatibility and avoidance of hepatotoxicity. Besides, G2111 encapsulated in A6-NP was only released less than 5% in 12 h, which may avoid off-target cell toxicity before entering into cancer cells. A6 peptide modification could significantly enhance uptake within a short time. Moreover, A6-NP continues to exert the broad anticancer spectrum of Hsp90 inhibitors and displays remarkable targeting ability and anticancer efficacy both in hematological malignancies and solid tumors (with colon tumors as the model cancer) both in vitro and in vivo. Overall, A6-NP, as a simple, biomimetic and active dual-targeting (CD44 and HSP90) nanomedicine, displays high potential for clinical translation.

Circular RNAs (circRNAs) are vital in many physiological and pathological events. Compared to the other processes of circRNA metabolism, circRNA degradation is less understood. Through RNAi screening and further characterization, RNAseK and lysosome are identified as circRNA degradation modules in metazoan, and RNase 1 as a lysosomal circRNA endoribonuclease in mammals. RNAseK and lysosome function synergistically, with RNAseK degrading circRNAs outside and lysosome degrading those inside the organelle. Mutations of RNAseK- or lysosome-sensitive sites in in vitro synthesized circRNA reporters lead to higher expression levels, and simultaneous mutations of both sensitive sites result in further increase. Under stress stimuli such as heat shock and ER stress, significantly decreased global circRNA levels are observed in Caenorhabditis elegans. RNAseK deficiency or lysosome inhibition diminishes the decreases in circRNA levels and impedes the induction of stress response, suggesting that circRNA degradation by RNAseK and lysosome plays protective roles in C. elegans stress response.

This study evaluated the prognostic performance of the 2023 International Federation of Gynecology and Obstetrics (FIGO) staging system in Chinese uterine serous carcinoma (USC) patients and defined the disease's molecular landscape. Retrospective analysis classified USC patients by both 2009 and 2023 FIGO systems; staging migration was analyzed via cross-tabulation and Sankey diagrams. Molecular classification used the Proactive Molecular Risk Classifier for Endometrial Cancer (ProMisE) algorithm, with human epidermal growth factor receptor 2 (HER2) expression assessed by immunohistochemistry (IHC). Progression-free survival (PFS) and overall survival (OS) were analyzed with Kaplan-Meier curves and log-rank tests; the 2023 system's prognostic ability was validated via time-dependent receiver operating characteristic (ROC) curves, Harrell's C-index, and Akaike's Information Criterion (AIC). The 2023 system induced significant staging migration, mainly from IA/IB to IC (endometrium-confined tumors) or IIC (myometrial invasion). This reclassification achieved better prognostic stratification, shown by higher C-index and lower AIC than the 2009 system. Molecular profiling clarified subtype distribution, and HER2 overexpression was confirmed in substantial subsets, highlighting therapeutic relevance. The 2023 FIGO staging system provides superior prognostic stratification for Chinese USC patients compared to the 2009 version.

High-grade serous ovarian carcinoma (HGSOC) remains the most lethal gynecologic malignancy, with platinum resistance posing the major therapeutic barrier. Malignant ascites, a hallmark of advanced HGSOC, correlates with chemoresistance and poor prognosis, yet the contribution of ascites circRNAs in this process remains obscure. Here, we identify ASCOR, a circRNA upregulated in ascites small extracellular vesicles (sEVs) from platinum-resistant HGSOC patients, predicts poor survival. ASCOR promotes platinum resistance in vitro and in vivo by enhancing cell viability, reducing apoptosis, and alleviating DNA damage, with effects also transferred via sEVs. Mechanistically, ASCOR binds RPA1 and facilitates its NUP153-dependent nuclear translocation, which recruits the RNA helicase DDX18, suppressing R-loop formation. Meanwhile, the ASCOR-RPA1-DDX18 axis activates PI3K/Akt signaling to promote platinum resistance. Targeting ASCOR with antisense oligonucleotides reverses chemoresistance in mouse models. Our study unveils an ascites circRNA-driven pathway underlying platinum resistance in HGSOC, suggesting targeting ASCOR is a promising strategy for HGSOC treatment.

e17505 Background: The clinicopathological profile and prognostic factors of cervical clear cell adenocarcinoma unrelated to diethylstilbestrol exposure are not well characterized in Chinese population. Methods: This multicenter retrospective study was conducted to analyze the characteristics of cervical clear cell adenocarcinoma patients from 10 medical centers. 97 patients surgically diagnosed with primary cervical clear cell adenocarcinoma were collected from 2010 to 2022. A comprehensive clinicopathological profile of cervical clear cell adenocarcinoma patients was depicted. The association between clinicopathological characteristics and prognostic factors was also assessed. Results: The peak age prevalence occurred in the 46-55 years group (n = 97), with the human papillomavirus (HPV)-negative rate of 85.1%. All surgically treated patients had FIGO stages IA-IIA or IIIC and received adjuvant radiotherapy and/or chemotherapy. With a median follow-up of 52 months (interquartile range, 36-70 months), multivariable analysis identified ovarian metastasis (HR = 3.89, P = 0.027 ) and lymph node metastasis (HR = 4.26, P = 0.004 ) as independent risk factors for overall survival. Multivariable analysis also demonstrated that lymph node metastasis (HR = 3.15, P = 0.005 ) remained a significant predictor for progression-free survival. Conclusions: This retrospective multicenter study characterized clinicopathological features of cervical clear cell adenocarcinoma and identified prognostic factors. Univariate and multivariate analysis of factors associated with overall survival. Variables Univariate HR (95%CI) P-value Multivariable HR (95%CI) P-value Tumor size >4 cm 3.85 (1.15–12.91) 0.029 3.13 (0.80–12.24) 0.102 DSI >2/3 8.30 (1.81–38.02) 0.006 3.36 (0.62–18.22) 0.161 Ovarian metastasis (Yes) 3.09 (1.03–9.39) 0.047 3.89 (1.17–12.98) 0.027 LVSI (Yes) 4.47 (1.80–11.07) 0.001 0.49 (0.26–3.58) 0.481 LNM (Yes) 4.80 (1.94–11.86) 0.001 4.26 (1.58–11.45) 0.004 Surgical approach (ARH) 1.57 (0.36–6.79) 0.548 - - HR, hazard ratio; 95%CI, 95% confidence interval; DSI, depth of stromal invasion; LVSI, lymphovascular space invasion; LNM, lymph node metastasis; ARH, abdominal radical hysterectomy; LRH, laparoscopic radical hysterectomy.

Pregnancy disorders in patients with autoimmune diseases or viral infections are often associated with an excessive response of type I interferons. We identify radical S-adenosyl methionine domain containing 2 (RSAD2) as a pathogenic interferon-stimulated gene (ISG) associated with pregnancy complications in systemic lupus erythematosus (SLE). The increased expression of RSAD2 mainly occurs in macrophages and structural cell populations at the maternal-fetal interface of pregnant patients with SLE. The elevation of RSAD2 leads to the accumulation of diacylglycerol lipids in the placenta, impairing the necessary vascular development for the fetus. Depletion of Rsad2 in pregnant mice models exposed to type I interferon inducers significantly reduces lipid accumulation, vascular injury, and embryo development disorders. An RSAD2 inhibitor, L-chicoric acid (LCA), alleviates lipid accumulation and vascular damage, improving pregnancy outcomes in SLE-induced and spontaneous mouse models. This study proposes the potential of targeting RSAD2 to improve pregnancy outcomes in individuals with heightened type I interferon response.

Background: The prognostic value of germline BRCA1/2 mutations ( gBRCA1/2m ) in ovarian cancer is controversial, and the clinical implications of specific mutation domains within BRCA1/2 remain underexplored. This study aimed to investigate the impact of distinct gBRCA1/2m domains on survival outcomes in patients with ovarian cancer. Methods: This multicenter retrospective study, conducted between 2010 and 2022 at three major academic centers in China, analyzed 313 patients with epithelial ovarian cancer with pathogenic gBRCA1/2m . We evaluated associations between gBRCA1/2m domains and clinical outcomes including progression-free survival (PFS) and platinum-free interval. Results: Patients who received platinum-based chemotherapy without maintenance therapy with BRCA1 C-terminal domain 1 (BRCT1) mutations showed significantly prolonged PFS (37.8 vs . 22.6 months; hazard ratio [HR], 0.43; 95% confidence interval [CI], 0.19–0.99; P = 0.042), whereas those with Really Interesting New Gene (RING) mutations had shorter PFS (13.1 vs . 23.2 months; HR, 1.82; 95% CI, 0.89–3.72; P = 0.097). In the subgroup of gross residual disease, RING mutations were significantly associated with reduced PFS (12.9 vs . 21.8 months; HR, 3.25; 95% CI, 1.29–8.18; P = 0.008). A higher likelihood of primary platinum-refractory disease (odds ratio, 7.78; 95% CI, 1.25–48.31; P = 0.028) was observed. Across all mutation locations, poly (ADP-ribose) polymerase inhibitor (PARPi) maintenance therapy demonstrated benefits, notably for patients with BRCA1 RING mutations (HR, 0.10; 95% CI, 0.01–0.84; P = 0.010) and BRCA2 RAD51-binding domain (RAD51-BD) mutations (HR, 0.29; 95% CI, 0.11–0.79; P = 0.010). Conclusions: BRCA1 BRCT1 mutations are associated with improved prognosis following platinum-based chemotherapy, whereas BRCA1 RING domain mutations are linked to a heightened risk of primary platinum-refractory disease. Our findings underscore the need for complete resection with no gross residual disease in patients harboring RING mutations. Furthermore, PARPi maintenance therapy exhibits variable efficacy based on mutation location, with BRCA1 RING and BRCA2 RAD51-BD mutations conferring significant benefits.

Ovarian cancer brain metastases (OCBM) are rare and have poor prognosis, with limited clinical management guidelines. This study aimed to identify prognostic factors and optimal treatment approaches for patients with OCBM. We conducted a retrospective multicenter analysis of patients with OCBM from 12 hospitals in China from May 2010 to May 2022. The primary outcomes were overall survival (OS) and brain metastasis–specific survival (BMSS). Kaplan–Meier and Cox regression analyses were used to assess treatment outcomes and identify prognostic risk factors. In total, 129 patients with OCBM were included. The median interval from ovarian cancer diagnosis to brain metastasis (BM) was 25.74 (range: 0–103.1) months. Headache attributed to BM was the most common presenting symptom, reported in 64 (49.6

e17541 Background: Ovarian cancer brain metastases (OCBM) are rare and have poor prognosis, with limited clinical guidelines. This study aims to identify prognostic factors and optimal treatment strategies for OCBM patients. Methods: We conducted a multicenter retrospective analysis of OCBM patients from 12 hospitals in China, spanning from May.1 2010 to May.31 2022. The primary endpoints were overall survival (OS) and brain metastasis-specific survival (BMSS). Kaplan-Meier and Cox regression analyses were used to assess treatment outcomes and identify risk prognostic factors. Results: A total of 129 OCBM patients were included. The median interval from ovarian cancer diagnosis to brain metastasis (BM) was 25.7 months (range: 0-102.9). Intracranial hypertension was the most common primary symptom reported in 64 (49.6%) patients, followed by paralysis in 23 (17.8%) patients. The cerebellum and brainstem were the most frequent sites of BM (36 patients, 27.9%), followed by the frontal lobe (27 patients, 20.9%). For BMSS-specific analysis, multiple BM lesions (HR 2.060, 95% CI: 1.308-3.244, p=0.002), intracranial hypertension (HR 1.765, 95% CI: 1.049-2.968, p=0.032), and relapse lines before BM diagnosis (HR 2.060, 95% CI: 1.308-3.244, p=0.002) as independent factors associated with poorer BMSS. Subgroup Kaplan-Meier analysis showed that patients with a single BM lesion had significantly better outcomes with stereotactic radiosurgery (SRS) compared to those with multiple lesions. Conclusions: This study identifies key prognostic factors for advanced OCBM patients, particularly intracranial hypertension, and the number of relapse-lines prior to BM diagnosis. Furthermore, it provides evidence that patients with a single BM lesion may benefit from SRS, whereas those with multiple lesions require more tailored treatment strategies. Univariate analysis of treatments for brain metastases. BM treatment OS BMSS Hazard ratio 95% CI P value Hazard ratio 95% CI P value Chemotherapy or support care Ref. Ref. WBRT ± Chemotherapy 0.78 0.412-1.477 0.446 0.652 0.334-1.274 0.652 Craniotomy ± Radiation/Chemotherapy 0.377 0.179-0.792 0.010 0.407 0.191-0.867 0.020 SRS ± Chemotherapy 0.270 0095-0.762 0.013 0.274 0.095-0.787 0.016 Abbreviations: BM: Brain metastasis; OS: Overall survival; BMSS: Brain metastases-specific survival; WBRT: Whole-brain radiotherapy; SRS: Stereotactic radiosurgery.

Background: Vascular endothelial growth factor receptor-3 (VEGFR-3) plays an indispensable role in lymphangiogenesis. Previous findings suggest that blocking the VEGFR-3 signaling pathway can inhibit lymph node metastasis effectively, thus reducing the incidence of distant metastasis. The development of new VEGFR-3-targeting drugs for early detection and effective treatments is, therefore, urgently required.Methods: In vitro biopanning of a phage-displayed peptide library was used to identify specific peptides binding to the extracellular domain of VEGFR-3. We obtained a novel VEGFR-3-targeting peptide, TMVP1 (LARGR). Our combined immunofluorescence and radiolabeling studies revealed that FITC-TMVP1 and 99mTc-labeled TMVP1 specifically accumulated in VEGFR-3-positive lymphatic vessels of tumors after intravenous administration in tumor xenograft models in vivo. To enhance the therapeutic efficacy of anticancer drugs, TMVP1 was fused to a proapoptotic peptide, D(KLAKLAK)2.Results: The fusion peptide strongly inhibited tumor lymphangiogenesis in vitro and in vivo and specifically suppressed lung metastasis in a 4T1 breast cancer xenograft model. The accumulation of the TMVP1 in lymphatic vessels was specific.Conclusions: Our results suggest that TMVP1 is a potential therapeutic strategy for developing new diagnostic tracers or alternative anticancer agents for tumor lymphangiogenesis and lymphatic metastasis.

High-grade serous ovarian cancer (HGSOC) is the most lethal type of gynecological cancer, and platinum-resistance is a serious challenge in its treatment. Long non-coding RNAs (lncRNAs) play critical regulatory roles in the occurrence and development of cancers. Here, using RNA sequencing of tumor small extracellular vesicles (sEVs) from HGSOC patients, the lncRNA CATED is identified as significantly upregulated in both tumors and tumor-derived sEVs in platinum-resistant HGSOC, and low CATED levels correlate with good prognosis. Functionally, CATED enhances cisplatin resistance by promoting cell proliferation and inhibiting apoptosis in vitro and in vivo. These effects could be transferred via CATED-overexpressing sEVs from donor cells and HGSOC tumor sEVs. Mechanistically, CATED binds to and upregulates DHX36 via PIAS1-mediated SUMOylation at the K105 site, and elevated DHX36 levels increase downstream RAP1A protein levels by enhancing RAP1A mRNA translation, consequently activating the MAPK pathway to promote platinum-resistance in HGSOC. Antisense oligonucleotide mediated knockdown of CATED reverse platinum-resistance in sEV-transmitted mouse models via the DHX36-RAP1A-MAPK pathway. This study newly identifies a sEV-transmitted lncRNA CATED in driving HGSOC platinum-resistance and elucidates the mechanism it regulates the interacting protein through SUMOylation. These findings also provide a novel strategy for improving chemotherapy in HGSOC by targeting CATED.

Rationale: Pyroptosis, an emerging form of programmed cell death, facilitates the release of tumor antigens and inflammatory factors, which can be leveraged to enhance the efficacy of immune checkpoint blockade (ICB) therapy. However, achieving high-efficiency induction of pyroptosis in cancer cells while minimizing toxicity remains a significant challenge. Methods: In this study, we designed a tumor-targeting peptide TMTP1-modified nanostructured lipid carrier (referred to as TP-NLC) with high loading capacities for gambogic acid (GA) and indocyanine green (ICG). The TMTP1, identified by our research team for its tumor-targeting capabilities, was conjugated to the nanocarrier surface using "click chemistry" to improve the drug delivery efficiency to tumor tissues. The TP-NLC nanocarrier was thoroughly characterized with respect to its morphological attributes, photostability, tumor-targeting capabilities, ability to induce pyroptosis, reactive oxygen species (ROS)-responsive behavior, and anti-tumor efficacy both in vitro and in vivo. Results: GA encapsulated within the TP-NLC nanocarrier, induced pyroptosis in tumor cells, and enhanced the efficacy of ICG-induced pyroptosis under laser irradiation by disrupting intracellular antioxidant systems, realizing that the combination of GA and ICG synergistically induced caspase-3/GSDME-mediated pyroptosis in a ROS-dependent manner. Tumor cells of pyroptosis released cellular contents and tumor antigens, which subsequently promoted the maturation of dendritic cells (DCs), enhanced intratumoral infiltration of CD8+ T cells, initiated systemic antitumor immune response, and augmented the efficiency of PD-1 blockade against both primary and metastatic tumors. Conclusion: The combination of GA and ICG therapy utilizing the constructed nanocarriers presents an attractive therapeutic strategy to trigger pyroptosis and potentiate PD-1 blockade therapy for cervical cancer chemo-immunotherapy.