Activation of cGAS-STING signaling in cancer cells requires cytosolic DNA produced by intrinsic or treatment-induced DNA damage. However, clinical efforts to exploit this pathway to improve immunotherapy have yielded limited success, highlighting gaps in understanding the link between DNA damage and immunotherapy. Here, we identify ubiquitination-directed cytosolic DNA degradation as a critical determinant for cGAS-STING activation following DNA damage. Mechanistically, the cytosolic DNA exonuclease TREX1 is degraded by the E3 ubiquitin ligase SPOP but is reversely stabilized by the deubiquitinase USP7. Cancer-associated SPOP mutations or USP7 overexpression elevate TREX1 levels, promoting cytosolic DNA degradation and impairing cGAS-STING-mediated immune activation. Notably, elevated USP7 expression correlates with reduced tumor-infiltrating lymphocytes and accelerated disease progression in patients undergoing chemoradiotherapy. Furthermore, USP7 inhibitors reduce TREX1 levels and restore immune responses following radiation. These findings elucidate the mechanisms linking DNA damage to immune activation and highlight USP7 inhibitors as potential enhancers of radioimmunotherapy.

The histological heterogeneity of primary tumors across the pan-cancer spectrum poses a formidable barrier to accurate lymph node metastasis assessment, often causing AI systems to make “overconfident errors” on rare variants that lead to missed diagnoses. To address this, we present UPATHLN, a unified diagnostic platform that synergizes a pathology foundation model-based encoder with a decoupled uncertainty estimation mechanism. We developed and validated the system using a large-scale multicentre dataset of 26,229 lymph nodes from 14 distinct primary origins. In internal validation, UPATHLN achieved an area under the curve (AUC) of 0.986. Crucially, the uncertainty module functioned as a decisive fail-safe: by flagging potential false-negative predictions for mandatory pathologist review, it intercepted all missed diagnoses, securing 100% conditional sensitivity across both the development and independent test cohorts—even for tumors from seven unseen primary origins. Concurrently, this mechanism reduced the review burden on negative lymph nodes by 73.2%. Ultimately, UPATHLN sets a new benchmark for safety-critical AI, demonstrating that explicitly modeling uncertainty is key to unlocking reliable, workload-efficient diagnostics at the pan-cancer scale.

Cervical cancer patients are at a high risk of tumour recurrence. Accurate delineation of the primary gross tumour volume (GTV) on pretreatment MR images is crucial for developing personalized prognosis evaluation and treatment. In this study, we collected pretreatment T2-weighted MR data from 306 cervical cancer patients (62 for testing) between 2011 and 2020. We developed a 3D MedNext-7L model for automatic GTV delineation and compared it with the nnUNetV2 model. Manual modifications were made to improve accuracy, and radiomics features were extracted to predict the risk of local recurrence within 5 years. Feature selection was performed using the least absolute shrinkage and selection operator algorithm, and modelling was conducted with the support vector machine algorithm. Additionally, we explored the potential of directly utilizing automatic delineation results for recurrence prediction. The Dice similarity coefficient and sensitivity for the MedNext-7L model in the test set were 0.812 ± 0.144 and 0.848 ± 0.128, respectively, surpassing those of nnUNetV2 at 0.803 ± 0.154 and 0.823 ± 0.155, respectively. Using manually refined GTVs, our model achieved AUC values of 0.875 and 0.811 in the fivefold cross-validation set and test set, respectively. On the basis of the automatically delineated GTVs, our model achieved AUC values of 0.786 and 0.730 in the fivefold cross-validation set and test set, respectively. The 3D MedNext-7L model outperforms nnUNetV2 in GTV delineation on pretreatment T2-weighted MR images. Rapid manual modifications combined with radiomics and machine learning techniques can effectively predict recurrence risk. The automatic delineation results show promise for direct application in recurrence prediction, indicating a feasible pathway towards a fully automated predictive system.

A substantial portion of patients experience radioresistance, which impedes clinical benefit. The radiation-induced 'protumor' immune response is previously demonstrated to limit antitumor efficacy. However, the detailed mechanism remains to be explored. In this study, we observe CXCR5+ monocytes are enriched in tumor upon radiation. CXCR5 expression on monocytes in host is induced by tumor-derived VEGF through PI3K/mTOR/HIF-1α axis. Local radiation enhances CXCL13 expression from tumor cells, a specific ligand of CXCR5, which leads to the recruitment of CXCR5+ monocytes. Tumor-infiltrating CXCR5+ monocytes induce radioresistance by inhibiting CD8+ T cells through PD-1/PD-L1 interaction. Moreover, radiation-induced GM-CSF promotes the differentiation of CXCR5+ monocytes toward M2-like macrophages. In contrast, inhibiting VEGFR signaling, neutralizing CXCL13 and GM-CSF, or blocking PD-L1 facilitates radiation-induced tumor control by abrogating CXCR5+ monocyte-mediated immunosuppression. Furthermore, the CXCR5+ and CD14+ populations are increased in patients with cancer following radiotherapy. Monocyte is increased in the peripheral blood of patients with progressive disease following radiotherapy. These findings suggest potential strategies for blocking the CXCR5/CXCL13 axis to improve radiotherapy efficacy.

Chemo-resistance is a major challenge in rectal cancer treatment. This study investigates the therapeutic potential of MEK inhibitors, cobimetinib and trametinib, in 5-fluorouracil (5-FU)-resistant rectal cancer cells. High-throughput drug screening identified these inhibitors as top candidates based on their selective drug sensitivity scores (sDSS). Both drugs exhibited dose-dependent cytotoxicity against rectal cancer cells while sparing normal epithelial cells and showed synergistic interactions with 5-FU. MEK inhibition disrupted redox homeostasis, increasing reactive oxygen species (ROS) and oxidative damage markers, including protein carbonyl and malondialdehyde (MDA), while also decreasing mitochondrial respiration, as evidenced by reduced oxygen consumption rates (OCR). Apoptotic induction was significantly reduced in mitochondrial respiration-deficient p⁰ cells, supporting the role of mitochondrial respiration in MEK inhibitor activity. Genetic MEK1/2 knockdown mimicked these effects, confirming MEK1/2 as a key regulator of oxidative stress and mitochondrial respiration. In vivo, cobimetinib and trametinib suppressed tumor growth in a chemo-resistant colorectal cancer xenograft model without significant toxicity, inducing oxidative stress and decreasing mitochondrial respiration. These findings highlight MEK inhibitors as promising candidates for overcoming chemo-resistance in rectal cancer by targeting oxidative stress and mitochondrial respiration.

Tumor metastasis is the primary cause of cancer treatment failure and mortality. Pregnant patients with cancer sometimes experience a poor prognosis and accelerated disease progression, yet the underlying mechanisms remain poorly understood. Here we show that pregnancy enhances tumor metastasis in female mice by elevating adenosine levels, which drives pre-metastatic niche remodeling. Myeloid cells in placental and uterine tissues exhibit increased expression of adenosine-generating enzymes, CD39 and CD73. The accumulated adenosine recruits neutrophils into the tissues of the pre-metastatic microenvironment and upregulates PD-L1 expression on these cells through the cAMP-PKA-NF-κB pathway, thereby suppressing CD8+ T cell function. Consistent with murine models, pregnant women exhibit elevated adenosine levels and increased PD-L1+ neutrophils in peripheral blood, hindering human T cell activation. Inhibiting the adenosine-PD-L1+ neutrophil axis reverses, at least partially, pregnancy-accelerated metastasis without affecting fetal development. These findings shed light on the mechanism of tumor metastasis mice during pregnancy and suggest potential therapeutic targets for treating cancers in pregnant patients.

Traditional cancer imaging modalities cannot achieve ideal diagnostic results. Fluorescence imaging is an emerging modality for tumor imaging because of its high selectivity and sensitivity. However, conventional imaging agents have some drawbacks, including significant photobleaching, low fluorescence quantum yield, and inadequate targeting specificity. Therefore, there is an urgent need for the development of new imaging agents. Viologen derivatives have been widely used in the optical field owing to their excellent water solubility and good optical properties. However, because of its propensity to readily obtain electrons and form free radicals, its biological toxicity is significant, restricting its further application in the field of biological staining. To solve these problems, this study incorporated phenyl viologen as the primary fluorescent structure, which is capable of directly forming a quinone structure in a single step, thereby mitigating the impact of free radicals on cells. Furthermore, the introduction of biotin further enhanced the targeting of the imaging agent, ensuring that it was delivered more precisely to the desired cellular locations. This dual approach not only minimizes the harmful effects of free radicals but also improves the specificity and efficiency of cellular imaging. Experimental results demonstrate that the developer exhibits high photostability, excellent biosafety, and outstanding biocompatibility. This study investigated the application of cationic viologen derivatives in living cell imaging, laying a foundation for the advancement of cationic viologen derivatives in the biological field.

Introduction XPO1 plays a crucial role in the nuclear export machinery, making it an attractive target for inhibiting nuclear-cytoplasmic transport in melanoma, where its overexpression is linked to unfavorable prognosis. However, XPO1 monotherapy has not demonstrated sufficient efficacy to be considered a first-line treatment option for melanoma. Objectives This research aimed to delve into the resistance mechanism of XPO1-targeting therapy in melanoma and fabricate a proteinoid microsphere which could target XPO1 and β-catenin to maximize the effect of XPO1 inhibitors. Methods Transcriptome sequencing was used to analyze the effects of XPO1 interference on the signaling pathways of melanoma. Nuclear-cytoplasmic protein separation, co-immunoprecipitation, and confocal microscopic analyses were conducted to clarify the resistance mechanism of XPO1 targeting therapy. A proteinoid microsphere named XPinβ was developed by co-assembling a specially designed XPO1 antagonistic peptide (XPin) and a β-catenin antagonist (Carnosic acid/CA). Cell model, mouse allograft and patient-derived xenograft (PDX) models were used to evaluate the antitumor effect of XPinβ. Results In our study, inhibition of XPO1 led to the nuclear accumulation of β-catenin, altered the nuclear-cytoplasmic localization of APC, and activated the Wnt/β-catenin signaling pathway. XPinβ was efficiently internalized into melanoma cells via macropinocytosis, achieving simultaneous inhibition of both XPO1 and β-catenin. As expected, XPinβ demonstrated robust anti-tumor efficacy in an allograft melanoma mouse model, with significantly superior therapeutic effects compared to monotherapy targeting XPO1 or CA treatment alone. Moreover, XPinβ effectively inhibited growth of patient-derived xenograft (PDX) tumors overexpressing XPO1, outperforming both CA and the commercially available XPO1 inhibitor KPT-330. Most importantly, XPinβ significantly suppressed pulmonary metastasis of melanoma while maintaining excellent biosafety. Conclusions This study demonstrates the enhanced efficacy of XPO1-targeted therapy through the inhibition of the Wnt/β-catenin signaling pathway and introduces XPinβ, a proteinoid microsphere with promising clinical translational potential for dual targeting therapy against melanoma involving both XPO1 and β-catenin.

Radiotherapy is a standard treatment for locally advanced cervical cancer. Based on the inherent characteristics of X-ray and γ-ray radiation techniques, we propose and investigate an innovative dual-modality radiation system incorporating both X-ray and γ-ray modalities. This study compares the quality of radiation treatment plans for patients with CC, generated using the dual-modality (TaiChi) system and a conventional LINAC X-ray system. Retrospective treatment plans using volumetric modulated arc therapy were selected for 12 patients with CC. Dual-modality plans were developed for these patients. All patients had the same prescription dose of 50 Gy/25 fractions for planning tumor volume (PTV) and 60 Gy/25 fractions for PTV-nd. The dose conformity index (CI) and gradient index (GI) were calculated to evaluate the dose coverage and drop for the target. The dose indices, Dmean, Dmax, and Dmin, were calculated for both the target and organs at risk (OARs). Both strategies generated dosimetrically acceptable plans. For both PTV-nd and PTV, the plans of TaiChi demonstrated higher CI values (PTV-nd: 0.68 ± 0.11 vs. 0.57 ± 0.22, P-value = 0.1375; PTV: 0.91 ± 0.02 vs. 0.90 ± 0.02, P-value = 0.1053) and lower GI values (PTV-nd: 280.62 ± 302.86 vs. 300.65 ± 317.83, P-value = 0.0094; PTV: 3.36 ± 0.22 vs. 3.66 ± 0.32, P-value = 0.0015) compared with the plans of the conventional LINAC system. For both PTV-nd and PTV, the TaiChi plans exhibited superior dosimetric indices compared to the LINAC plans. Specifically, the Dmax (78.27±5.20 Gy vs. 66.61±1.51 Gy) and Dmean (66.85±1.61 Gy vs. 63.22±1.03 Gy) of the TaiChi plans were significantly higher than those of the LINAC plans. Although the Dmin (61.06±0.52 Gy vs. 60.46±1.33 Gy) of the TaiChi plans was also higher than that of the LINAC plans, the difference was not statistically significant. Moreover, TaiChi plans significantly reduced radiation exposure to OARs compared to LINAC plans, demonstrating improved organ protection. This was particularly evident for the spinal cord PRV, with the TaiChi plans yielding a maximum dose of 23.61±2.80 Gy, compared to 29.85±3.23 Gy for the LINAC plans (P-value = 0.0005). The dual-modality system has the potential to enhance treatment outcomes in CC by optimizing tumor coverage while reducing toxicity. Future clinical trials should assess its impact on patient survival and treatment-related side effects to bridge the gap between dosimetric improvements and clinical application.

Defining an ERBB2 (HER2/neu) gene amplification status is critical to guiding human epidermal growth factor receptor 2 (HER2)-targeted therapy in breast cancer. Up to 40% of breast cancer patients are reported as having an immunohistochemistry (IHC) of HER2 2+ and requiring additional testing using fluorescence in situ hybridization to confirm the results. This paper aims to establish an automatically weighted calibration deep learning (AWCDL) algorithm to predict ERBB2 amplification based on IHC images. In this study, we applied IHC HER2 2+ images from 1,073 breast cancer patients at three cancer centers in China and extracted 376,099 tiles. Among these, 269,664 tiles were used for internal and external validation. The designed AWCDL consists of two steps. In Step 1, the internal validation achieved an accuracy of 89%, with a specificity of 0.89 and a sensitivity of 0.89. The external validation in the two other centers showed an average accuracy of 85%, with a specificity of 0.86 and a sensitivity of 0.82. In Step 2, the model achieved higher accuracy for the slides predicted as negative in Step 1 by automatically calibrating the weight. Collectively, these results suggest that this AWCDL model has successfully proved useful as an alternative method to fluorescence in situ hybridization for assessing the ERBB2 amplification status in breast cancer.

Periodontitis is a destructive disease that causes irreversible damage to periodontal supporting tissues, posing a serious threat to patients' oral health. Given the insidious nature of periodontal tissue destruction, early diagnosis of periodontitis is crucial. Salivary hydrogen sulfide (H2S) has been identified as a potential biomarker for periodontitis monitoring. However, existing detection methods struggle to precisely locate the source of H2S release within the oral cavity, making it difficult to accurately identify affected areas and significantly hindering early diagnosis and treatment of periodontitis. This study reports a visual aggregation-induced emission (AIE) fluorescent probe (TTN), which exhibits high sensitivity and rapid responsiveness to H2S, along with dual-signal response capabilities in color and fluorescence. Based on this probe, a highly reliable and practical TTN film sensor capable of efficiently monitoring H2S is developed. The film can be used for real-time diagnosis of periodontitis, allowing precise identification of diseased tooth areas through visible color and fluorescence changes. Additionally, the film demonstrates efficient reactive oxygen species (ROS) generation, inhibiting the growth of periodontitis-causing bacteria and showcasing therapeutic potential. This integrated diagnostic and therapeutic design not only provides a novel approach for monitoring H2S as a biomarker but also opens new avenues for precise diagnosis and treatment of periodontitis.

Aim:To observe the effect of vitamin C on Kidney renal clear cell carcinoma (KIRC) and investigate its mechanism. Methods and Results:Firstly, 29 vitamin C direct target proteins (DPTs) were identified by Drug Bank 5.0, and the protein-protein interaction (PPI) network and signaling pathways of vitamin C DPTs were analyzed. The results showed that vitamin C was not only related to KIRC, but also to the HIF-1 pathway. Meanwhile, the top 300 highly expressed genes of KIRC were obtained by GEPIA. Next, We compared the genes of four vitamin C targets in the PPI network with highly expressed genes in KIRC. Interestingly, these common genes are also involved in HIF-1 pathway. Additionally, we utilized RNA-Seq technology to explore the differentially expressed genes in KIRC with vitamin C compared to those not intervened. We observed that these differentially expressed genes exhibited a close association with hypoxia. Finally, we observed the inhibitory effect of Vitamin C on KIRC by Cell Counting Kit-8 (CCK8) assay, real-time quantitative PCR, Western blotting, flow cytometry, and colony formation assay, and confirmed that Vitamin C inhibits the growth of KIRC cells through the HIF-1 pathway. Conclusion:Through bioinformatics analyses, we identified the molecular mechanism of vitamin C's role in KIRC and verified it through a series of experiments. Combined bioinformatics analysis will play an important role in future drug-disease interaction studies.

BACKGROUND:Radiotherapy (RT) is a cornerstone of cancer treatment alongside surgery. However, thoracic RT carries a substantial risk of Radiation-Induced Heart Disease (RIHD), characterized by acute activation of inflammatory pathways that evolve into chronic pathological cascades. This study aimed to elucidate the molecular mechanisms underlying cardiac responses to irradiation, with the goal of establishing a theoretical basis for RIHD prevention and providing new insights for further investigation. METHODS:DNA damage in cardiomyocytes after irradiation was assessed by immunofluorescence. Cell proliferation was evaluated using the CCK-8 assay, while apoptosis and cell cycle distribution were analyzed by flow cytometry. Protein expression levels of NRG-1, ErbB4, ErbB2, and p53 were detected by Western blotting. Serum cardiac injury biomarkers were quantified by enzyme-linked immunosorbent assay (ELISA). Myocardial inflammation and fibrosis were evaluated histologically using hematoxylin and eosin (H&E) staining and Masson's trichrome staining. RESULTS:X-ray irradiation induced significant DNA damage, cell cycle arrest, apoptosis, inflammatory activation, and p53 upregulation in cardiomyocytes. In parallel, irradiation suppressed the NRG-1/ErbB4/ErbB2 signaling pathway, thereby reducing proliferation. Histological analysis confirmed that irradiation promoted myocardial inflammation and fibrosis. The overexpression of NRG-1 conferred cardioprotection after radiation by mitigating DNA damage, facilitating cell cycle progression, and inhibiting apoptosis and inflammation, accompanied by the suppression of p53 and induction of ErbB4/ErbB2 expression. Serum cardiac injury biomarkers in irradiated rats exhibited a significant dose-dependent elevation following thoracic RT. CONCLUSION:The NRG-1/ErbBs signaling pathway mitigates p53 activation and, through this mechanism, promotes DNA repair, facilitates cell cycle progression, inhibits apoptosis, and reduces inflammation. Collectively, these effects enhance cardiomyocyte proliferation and confer cardioprotection against radiation-induced injury.

Background: Colon cancer is a malignant tumor with high malignancy and a low survival rate whose heterogeneity limits systemic immunotherapy. Transforming growth factor-β (TGF-β) signaling pathway-related genes are associated with multiple tumors, but their role in prognosis prediction and tumor microenvironment (TME) regulation in colon cancer is poorly understood. Using bioinformatics, this study aimed to construct a risk prediction signature for colon cancer, which may provide a means for developing new effective treatment strategies. Methods: Using consensus clustering, patients in The Cancer Genome Atlas (TCGA) with colon adenocarcinoma were classified into several subtypes based on the expression of TGF-β signaling pathway-related genes, and differences in survival, molecular, and immunological TME characteristics and drug sensitivity were examined in each subtype. Ten genes that make up a TGF-β-related predictive signature were found by least absolute shrinkage and selector operation (LASSO) regression using colon cancer data from the TCGA database and confirmed using a Gene Expression Omnibus (GEO) dataset. A nomogram incorporating risk scores and clinicopathologic factors was developed to stratify the prognosis of patients with colon cancer for accurate clinical diagnosis and therapy. Results: Two TGF-β subtypes were identified, with the TGF-β-high subtype being associated with a poorer prognosis and superior sensitivity to immunotherapy. Mutation analyses showed a high incidence of gene mutations in the TGF-β-high subtype. After completing signature construction, patients with colon cancer were categorized into high- and low-risk subgroups based on the median risk score of the TGF-β-related predictive signature. The risk score exhibited superior predictive performance relative to age, gender, and stage, as evidenced by its AUC of 0.686. Patients in the high-risk subgroup had higher levels of immunosuppressive cell infiltration and immune checkpoints in the TME, suggesting that these patients had better responses to immunotherapy. Conclusions: Patients with colon cancer were divided into two subtypes with different survival and immune characteristics using consensus clustering analysis based on TGF-β signaling pathway-related genes. The constructed risk prediction signature may show promise as a biomarker for evaluating the prognosis of colon cancer, with potential utility for screening individuals for immunotherapy.

Background: Concurrent chemoradiotherapy is the preferred treatment for stage IVB cervical cancer; however, some patients experience a poor prognosis. The prognostic significance of body composition indicators, including visceral obesity, has been extensively investigated in patients with cancer. This study aimed to assess the impact of body composition indicators, specifically pretreatment fat content, on the survival outcomes of patients with stage IVB cervical cancer. Methods: We retrospectively analyzed clinical information from patients diagnosed with stage IVB cervical cancer between 2010 and 2018. We measured visceral obesity (visceral-to-subcutaneous adipose tissue area ratio [VSR]) and skeletal muscle index (SMI) on pretreatment computed tomography (CT) images. We evaluated the impact of these body composition parameters on the prognosis of patients with cervical cancer. Results: Overall, 116 patients were included, 81 of whom had complete clinical and imaging information. Based on the cut-off values from X-tile analysis, we categorized patients into high and low VSR and SMI groups. The overall survival (OS) rate of patients with a high VSR was significantly higher than that of patients with a low VSR

BackgroundThe activation of the DNA damage response (DDR) heavily relies on post-translational modifications (PTMs) of proteins, which play a crucial role in the prevention of genetic instability and tumorigenesis. Among these PTMs, palmitoylation is a highly conserved process that is dysregulated in numerous cancer types. However, its direct involvement in the DDR and the underlying mechanisms remain unclear.MethodsCRISPR-Cas9 technology was used to generate the PORCN KO and PORCN NLS KO cell lines. The effects of PORCN NLS in the DDR were verified by colony formation assays, MTT assays, the DR/EJ5 homologous recombination/non-homologous end-joining reporter system, xenograft tumor growth and immunofluorescence. Mechanisms were explored by mass spectrometry, acyl-biotin exchange (ABE) palmitoylation assay, Click-iT assay, cell subcellular fractionation assay, Western blot analysis, and in vivo and in vitro co-immunoprecipitation.ResultsIn this study, we introduce evidence that Porcupine (PORCN) is an integral component of and plays a critical role in the DDR. PORCN deficiency hampers nonhomologous end joining (NHEJ) and highly sensitizes cells to ionizing radiation (IR) both in vitro and in vivo. We also provide evidence that PORCN possesses a nuclear fraction (nPORCN) with S-acyltransferase activity, unlike its membrane-bound O-acyltransferase in the endoplasmic reticulum. Furthermore, we show that nPORCN is necessary for the successful activation of NHEJ. Using mass spectrometry, we reveal the existence of an nPORCN complex and show that nPORCN mediates the S-palmitoylation of XRCC6/Ku70 at five specific cysteine sites in response to IR. Mutation of these sites causes a substantial increase in radiosensitivity and delays NHEJ. Additionally, we present evidence that nPORCN-dependent Ku70 palmitoylation is required for DNA-PKcs/Ku70/Ku80 complex formation.ConclusionOur findings underscore the crucial role of nPORCN-dependent Ku70 S-palmitoylation in the DDR.

1Center of Digestive Endoscopy, Shaanxi Provincial Cancer Hospital, Xi’an, China 2The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi’an Jiaotong University, Xi’an, China 3Department of Medical Oncology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an, China 4Department of Radiation Oncology, The First Affiliated Hospital of Xi’an Jiaotong University, Xi’an Jiaotong University, Xi’an, China 5Department of Epidemiology, Shaanxi Provincial Cancer Hospital, Xi’an, China