Recent studies have demonstrated that immune responses are subject to epigenetic regulation. However, the functional roles of RNA N6-methyladenosine (m6A) modification in infiltrating cells within the tumor microenvironment (TME) remain incompletely understood. To systematically characterize m6A modification patterns, we conducted a comprehensive analysis across 1,195 colorectal cancer samples, focusing on 27 m6A regulators. We further associated the three identified modification patterns with the characteristics of immune cell infiltration in the TME. To quantify m6A modification patterns at the individual tumor level, we developed an m6A score using principal component analysis (PCA). Analysis of immune cell infiltration in the TME revealed that the three distinct m6A modification patterns were significantly associated with three tumor immune phenotypes: immune-excluded, immune-inflamed, and immune-desert. Furthermore, univariate and multivariate Cox regression analyses indicated that the m6A score served as an independent and reliable prognostic biomarker for colorectal cancer (CRC) patients. Collectively, these findings suggest that m6A modification is associated with immune modulation and correlates with TME heterogeneity. Characterization of m6A modification patterns in CRC patients may enhance our understanding of immune cell infiltration within the TME and inform immunotherapeutic strategies.

Major Depressive Disorder (MDD) is a highly disabling psychiatric condition. The diagnostic and therapeutic challenges associated with MDD stem from an incomplete understanding of its pathogenesis, particularly the complex cellular interactions and the influence of peripheral factors on the central nervous system (CNS). Current metagenomics and single-cell studies are largely confined to individual omics layers. This study integrated multi-cohort gut metagenomic data with cortical single-nucleus transcriptomics to elucidate gut-brain axis regulatory mechanisms in depression. We observed significant enrichment of Streptococcus in MDD patients, accompanied by reprogramming of microbial pathways related to neural signaling and lipid metabolism. Cortical analysis revealed reduced astrocyte proportions with functional impairment, and increased proportions of L5/6_THEMIS_1 and ID2_PLP1 neuron subtypes exhibiting metabolic stress signatures. Diagnostic models based on microbial signatures and astrocyte markers achieved AUCs of 0.914 and 0.854, respectively. Cross-omics correlation analysis demonstrated a significant association between microbial and cortical pathways (R = 0.796) and identified five core functional modules. This study constructs a molecular network of gut-brain axis regulation in MDD, revealing multi-dimensional functional coupling between gut microbiota and cortical cells, and providing evidence for how peripheral microbes influence central cellular function.

BackgroundTo evaluate the effect of Icariin (ICA) on AOM/DSS-induced colorectal cancer (CRC) in mice as well as CRC cells, and to explore the underlying molecular mechanism.MethodsThe therapeutic efficacy of ICA against AOM/DSS-induced CRC was evaluated by detecting tumor growth and pathological alterations. Biomarkers related to Wnt/β-catenin and mitochondrial apoptosis were tested, and cell experiments were performed to validate the regulatory correlation between ICA and the Wnt/β-catenin pathway in CRC cells.ResultsICA significantly reduced tumor number, size and ameliorated pathological damage in AOM/DSS-induced mice by regulating the mitochondrial-mediated apoptosis of colon tissue. ICA’s regulation of mitochondrial-mediated apoptosis was confirmed to involve the Wnt/β-catenin signaling pathway. In vitro, ICA restrained proliferation, migration and invasion of SW1463 and HCT116 cells, decreased mitochondrial membrane potential and changed the expression of pathway-related genes.ConclusionThe study demonstrated that ICA promoted mitochondrial-mediated apoptosis by regulating the Wnt/β-catenin signaling pathway, suggesting that ICA could serve as a potential treatment for CRC.

Purpose:Microbial communities have emerged as crucial regulators in the initiation and progression of thyroid cancer. However, most studies focus on single microbial sources, and the interplay between microbes across different ecological niches and their impact on thyroid carcinogenesis are largely unknown. Patients and Methods:In this study, we collected tissue, oral, and fecal samples from 32 patients with benign thyroid nodules (BTN) and 32 patients with papillary thyroid carcinoma (PTC). The oral and fecal samples were subjected to 16S rRNA sequencing, while the tissue samples were analyzed using 5R 16S sequencing to comprehensively characterize the microbial communities. Results:Clustering analysis using the Dirichlet Multinomial Mixture model with Laplace approximation identified two distinct oral microbial community types (O_1 and O_2) and three fecal types (F_1, F_2, and F_3). Microbial diversity patterns in thyroid tissues mirrored those observed in oral and fecal samples, suggesting potential microbial translocation or systemic interactions. Inflammatory markers were significantly elevated in PTC patients relative to BTN controls. Notably, the genus Veillonella, a potential anti-tumor biomarker, was significantly reduced in PTC samples across niches. Conclusion:This study highlights the pivotal role of oral and intestinal microbiota in PTC development, emphasizing the interplay between microbial composition, inflammatory processes, and immune regulation in tumor progression. The discovery of Veillonella as a potential anti-tumor microbe, along with evidence of microbial translocation, opens new possibilities for targeted therapeutic strategies.

Breast cancer (BC) is the most common malignancy among women, with its progression and prognosis significantly influenced by the tumor microenvironment (TME). Age-related differences in TME composition lead to distinct tumor behaviors: young patients (≤ 40 years) exhibit aggressive tumors, while elderly patients (> 70 years) experience immunosenescence and reduced therapy responses. We performed single-cell RNA sequencing (scRNA-seq) analysis on tumors from 10 breast cancer patients (5 ≤ 40 years, 5 ≥ 70 years), encompassing 33,664 high-quality cells. After cell annotation and batch correction, malignant epithelial cells were identified using inferCNV. We applied pseudotime trajectory analysis, pathway enrichment, and cell-cell communication profiling to investigate age-specific TME dynamics. Survival relevance was assessed using a GEO cohort (GSE20685) of young breast cancer patients, and immunohistochemical staining was performed on clinical tumor and fibroadenoma tissues to validate protein-level expression of key ISGs. In young patients, malignant epithelial cells showed gradual upregulation of interferon-stimulated genes (ISGs) such as IFI44, IFI44L, IFIT1, and IFIT3 along the pseudotime trajectory, suggesting their involvement in early tumorigenesis. High expression of these ISGs was significantly associated with poor overall survival in a young BC cohort (GSE20685). Immunohistochemical validation further confirmed elevated IFIT3 protein levels in young tumor tissues. In contrast, elderly patients had a TME enriched in macrophages and fibroblasts, with activation of immunosuppressive pathways (e.g., SPP1, COMPLEMENT). Our integrative analysis identifies ISGs as key transcriptional drivers of tumorigenesis in young breast cancer, with potential prognostic and therapeutic value. Despite limited sample size, the combination of single-cell transcriptomics, clinical survival data, and protein-level validation provides robust evidence of age-specific TME remodeling. These findings support the development of age-tailored immunotherapy strategies targeting interferon signaling in young patients and immune checkpoint pathways (e.g., LAG3, CTLA4) in elderly individuals.

BackgroundGynecomastia, characterized by benign proliferation of male breast glandular tissue, is a prevalent condition with complex etiologies. However, the absence of effective in vitro models has hindered mechanistic investigations and therapeutic development.MethodsIn this study, we established and characterized organoids derived from the breast tissues of six male gynecomastia patients, including physiological, idiopathic, and hormone-related subtypes. Organoid fidelity was evaluated using hematoxylin and eosin (H&E) staining, immunohistochemistry (IHC), immunofluorescence (IF), and quantitative PCR (qPCR), targeting a panel of lineage-specific and proliferative markers.ResultsThe organoids recapitulated key histological and molecular features of their corresponding source tissues, including epithelial architecture and expression of CK14, CK18, Ki67, and ERα. Marker expression was generally consistent between organoids and tissues at both the protein and transcriptional levels. Notably, ERα protein levels were reduced in organoids, while ESR1 mRNA expression remained stable, suggesting post-transcriptional regulation related to culture conditions.ConclusionOur study presents a practical and reproducible protocol for generating gynecomastia-derived organoids and highlights their utility as a disease-relevant platform for future research in male breast pathology and hormone-related mechanisms.

IntroductionIn thyroid carcinoma (TC), follicular thyroid carcinoma (FTC) represents the second most prevalent pathological type following papillary thyroid carcinoma. Notably, FTC exhibits a more aggressive clinical course and a higher propensity for distant metastasis. However, the underlying mechanisms governing the progression of FTC remain poorly understood. PMAIP1 is a gene implicated in various cancers and biological processes. Investigating the role and mechanism of PMAIP1 in FTC is crucial for enhancing our understanding of FTC and informing clinical treatment strategies. MethodsThis study examined the expression level of PMAIP1 in FTC through comprehensive analysis of databases, tumor tissues, and cell lines. Following the establishment of a stably transfected plasmid in cell lines, a series of functional assays and subcutaneous xenograft experiment were conducted to investigate the role of PMAIP1 in FTC. Additionally, transcriptome sequencing was employed to identify potential signaling pathways associated with PMAIP1. Mechanistic studies involved a series of rescue experiments to elucidate the regulatory mechanisms of PMAIP1 in FTC. ResultsPMAIP1 was found to be highly expressed in FTC, and its knockdown significantly inhibited the proliferation and metastasis of FTC cells both in vivo and in vitro. The results of transcriptome sequencing analysis indicated that PMAIP1 may influence the progression of FTC via the Wnt signaling pathway. Subsequent investigations revealed a direct correlation between PMAIP1 expression levels and those of Wnt3 and FOSL1 in FTC. A series of rescue experiments further substantiated the regulatory role of PMAIP1 on Wnt3/FOSL1 in FTC. DiscussionIn conclusion, our research demonstrated that PMAIP1 emerges as a novel pro-cancer factor in FTC, and its knockdown significantly inhibited the proliferation and metastasis of FTC both in vivo and in vitro. Mechanistically, PMAIP1 regulated FOSL1 by modulating the Wnt signaling pathway, thereby promoting FTC progression. Targeting PMAIP1 may present a promising therapeutic strategy for FTC.

IntroductionThe primary clinical challenge associated with follicular thyroid carcinoma (FTC) lies in accurately diagnosing the condition, particularly in distinguishing it with follicular thyroid adenoma (FTA) due to their overlapping cytomorphological features and sonographic characteristics.MethodsWhole exome sequencing (WES) techniques and Gene Expression Omnibus (GEO) database were utilized to analyze genomic difference between FTC and FTA, with a specific focus on immune-related genes. The hub genes were subjected to enrichment analysis, immune infiltration analysis, protein-protein interaction (PPI) analysis, and receiver operating characteristic (ROC) curve analysis. Then utilized quantitative real-time polymerase chain reaction (qRT-PCR) and immunohistochemistry (IHC) to validate the expression levels of PMAIP1 and PDGFRL at the cellular and tissue levels.ResultsThe findings of WES and bioinformatics analysis indicated that PMAIP1 and PDGFRL were potential mutated immune-related genes in FTC, in comparison to FTA, the expression of PMAIP1 is up-regulated in FTC while PDGFRL is down-regulated, demonstrating promising diagnostic efficacy. Enrichment analysis and immune infiltration analysis suggested that PMAIP1 and PDGFRL may serve as potential therapeutic targets for FTC. The results of the validation at both cellular and tissue levels indicated an up-regulation of PMAIP1 and a down-regulation of PDGFRL in FTC, consistent with the results from bioinformatics analysis. DiscussionIn conclusion, it is the first research to revealed PMAIP1 and PDGFRL as potential novel immunodiagnostic markers for FTC, shedding light on their potential biological significance in this context, and offering potential valuable clinical applications.

BACKGROUND Jejunal diverticula (JD) are rare clinical conditions that are typically incidentally detected and asymptomatic. When acute complications arise, surgical exploration may be necessary for accurate diagnosis and appropriate treatment. In this report, we present a case of multiple JD complicated by gastrointestinal bleeding and review the pathogenesis, diagnosis, and treatment of JD to increase clinician awareness of this condition. CASE SUMMARY A 70-year-old male patient with multiple JD presented with repeated massive gastrointestinal bleeding. The patient did not respond to symptomatic conservative treatment. Additional diagnostic investigations, including digestive endoscopy and abdominal angiography, did not reveal any relevant abnormalities. An exploratory laparotomy was subsequently performed, during which a segment of the bowel containing numerous diverticulum-like structures was surgically removed. Following successful discharge from the hospital, the patient did not experience any further episodes of gastrointestinal bleeding during subsequent follow-up. CONCLUSION Complications caused by JD are often difficult to diagnose, and surgical exploration is sometimes the most appropriate method.

Colorectal cancer (CRC) is a prevalent digestive system malignancy accompanied by peritoneal metastasis occurring in 7% of cases. Methyltransferase-like 3 (METTL3) promoted the progression of CRC whereas its function in peritoneal metastasis was incompletely understood. Here, we found that METTL3 was upregulated in peritoneal metastasis tissues of CRC patients compared with CRC tissues. By sequencing the mRNA of above tissues, we discovered that METTL3-mediated N6-methyladenosine (m6A) modification regulated the downstream target neurexin-3 (NRXN3). NRXN3 promoted CRC peritoneal metastasis in vivo. Mechanically, we further verified the specific methylation sites of NRXN3 mRNA modified by METTL3. Functional cellular assays demonstrated METTL3-mediated upstream regulation of NRXN3. We also demonstrated that YTHDC1 is necessary for the METTL3-mediated stabilization of NRXN3 mRNA. Taken together, this study established a METTL3-YTHDC1-NRXN3 m6A modification-dependent regulation system in CRC peritoneal metastasis, suggesting potential candidates for peritoneal metastasis treatment.

Objective:To explore the correlation between chromosome 8 open reading frame 76 (C8orf76) and cyclin-dependent kinase 4 (CDK4) and the potential predictive effect of C8orf76 and CDK4 on the prognosis of colorectal cancer (CRC). Methods:We constructed a protein-protein interaction network of C8orf76-related genes and analyzed the prognostic signatures of C8orf76 and CDK4. Clinicopathological features of C8orf76 and CDK4 were visualized using a nomogram. Results:C8orf76 and CDK4 levels were positively correlated in two independent human CRC cohorts ( n = 83 and n = 597). A consistent positive correlation was observed between C8orf76 and CDK4 expression in the CRC cell lines. The nomogram included prognostic genes (C8orf76 and CDK4) and pathological N and M stages. The concordance index (C-index) in our cohort was 0.776, which suggests that the ability of the indicators to predict the overall survival of patients with CRC in our cohort was strong. Conclusion:We found that C8orf76 was positively correlated with CDK4 in both the cohorts as well as in CRC cell lines. Therefore, C8orf76 and CDK4 can be used as potential biomarkers to predict the prognosis of CRC.

Immune checkpoint blockade (ICB) therapy demonstrated limited efficacy in colorectal cancer (CRC), and identification of intrinsic factors modulating immune suppression in CRC is an unmet need. Here, we revealed that Chromosome 8 open reading frame 76 (C8orf76) drives immunosuppression and is a molecular target to boost ICB therapy in CRC. C8orf76 is upregulated in primary CRCs compared to adjacent normal tissues in 2 independent CRC cohorts and its expression predicts poor patient survival. Intestine-specific C8orf76 knockin in mice exacerbated AOM/DSS-induced CRC, accompanied by increased intratumoral myeloid-derived suppressor cells (MDSCs) but reduced IFN-γ+ and granzyme B+ CD8+ T cells, inferring that C8orf76 promotes immunosuppression in CRC. Consistently, genetic depletion of C8orf76 augmented antitumour immunity in CT26 (MSS-CRC) and MC38 (MSI-H-CRC) allograft models. Integrated RNA-seq and ChIP-seq revealed that C8orf76 functions as a transcription factor to drive NDST1 expression, which activates the PI3K-Akt-NF-κB signaling cascade. Activated NF-κB in turn promotes the expression and secretion of CXCL1, a major chemoattractant for MDSCs. Consequently, C8orf76-induced CXCL1 mediates the recruitment of MDSCs via CXCR2 to antagonize functional CD8+ T cells in tumor immune microenvironment of CRC. Confirming this, targeting NDST1 or CXCR2 could reverse C8orf76 induced immunosuppression in vivo. Finally, we tested the translational value of C8orf76 using genetic ablation or vesicle-like nanoparticles (VNPs)-encapsulated C8orf76-siRNA. In line with our hypothesis, we demonstrated that targeting C8orf76 by genetic ablation or VNPs-encapsulated C8orf76-siRNA potentiated the anti-PD1 efficacy in both MSI-H and MSS CRC models. In summary, C8orf76 drives immunosuppression through a NDST1-CXCL1/CXCR2 axis, and targeting C8orf76 is a promising approach to boost ICB therapy efficacy in CRC. Hongyan Gou, Shang Guo, Xiaohong Wang, Xia Jiang, Chi Chun Wong, Huarong Chen, Chunxian Wei, Lingxue Shi, Zengren Zhao, Jun Yu. C8orf76 promotes colorectal tumorigenesis by promoting an immunosuppressive microenvironment and is a therapeutic target for boosting anti-PD-1 efficacy [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2025; Part 2 (Late-Breaking, Clinical Trial, and Invited Abstracts); 2025 Apr 25-30; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2025;85(8_Suppl_2):Abstract nr LB132.

Background:The prognosis of colorectal cancer (CRC) varies significantly across different immune subtypes. This study aimed to develop a risk prediction model incorporating the tumor immune microenvironment (TIME) to improve prognosis assessment and predict immunotherapy response in CRC patients, given the significant variability in clinical outcomes across different immune subtypes. Methods:CRC transcriptome data and corresponding clinical information were obtained from The Cancer Genome Atlas (TCGA) database. Univariate and multivariate Cox regression analyses were employed to identify m6A-related long non-coding RNAs (lncRNAs) (mRLs). A risk model was constructed using least absolute shrinkage and selection operator (LASSO) Cox regression and further validated through nomogram analysis, time-dependent receiver operating characteristic (ROC) curves, and Kaplan-Meier survival analysis. Differences in immune infiltration scores and clinical characteristics between low-risk group (LRG) and high-risk group (HRG) were also investigated. Results:An 11-mRL signature model was established based on their expression profiles in CRC and correlation with m6A regulatory factors. This model demonstrated strong predictive performance for OS, as confirmed by Kaplan-Meier analysis, ROC curves, and Cox regression. Notably, the HRG exhibited significantly higher infiltration of specific immune cells and elevated expression of immune checkpoints [programmed cell death protein 1 (PD-1), programmed death-ligand 1 (PD-L1), and cytotoxic T lymphocyte antigen 4 (CTLA4)] compared to the LRG. Furthermore, the two groups showed distinct responses to immunotherapy, suggesting potential utility in guiding immunosuppressant selection. A nomogram integrating m6A-immune signatures and clinicopathological variables was developed to individualize prognosis prediction. Conclusions:This study constructed an mRLs risk model that effectively predicts CRC prognosis and immune profiles, offering a potential tool for personalized therapeutic decision-making.

Background: Colorectal cancer (CRC) stands out as one of the most widespread and life -threatening malignancies globally. In recent years, significant attention has been directed towards the Yes -associated protein 1 (YAP1), identified as the central effector molecule in the Hippo signaling pathway, highlighting its crucial involvement in CRC development. Nevertheless, the role of hematopoietic SH2 domain -containing (HSH2D) in CRC, and its impact on tumor progression and resistance to chemotherapy by modulating YAP1, remains unclear. Therefore, this study aims to explore the role of HSH2D in CRC and assess its potential therapeutic significance. Methods: The study included the analysis of HSH2D expression levels in CRC tissues and cells by immunohistochemistry and western blot techniques. The influence of HSH2D on CRC cell proliferation and migration was examined using RNA interference and overexpression systems. The regulatory impact of HSH2D on YAP1 expression was assessed through real-time reverse transcriptase-polymerase chain reaction (RT-qPCR) and immunoblotting. In vitro cell experiments were conducted to unravel the mechanism by which HSH2D promotes CRC progression through YAP1. Lastly, the study investigated the role of HSH2D in modulating the sensitivity of CRC cells to cisplatin by chemosensitivity experiments. Results: The experimental findings demonstrated a notable elevation in HSH2D expression within both CRC tissues and cells. Overexpression of HSH2D was found to enhance the proliferation and migration capabilities of CRC cells, whereas silencing HSH2D had the opposite effect. Subsequent investigations unveiled that HSH2D upregulated YAP1 expression, and the tumorpromoting influence of HSH2D was, in part, dependent on YAP1. Furthermore, heightened HSH2D expression correlated with increased resistance of CRC cells to cisplatin. Conclusion: This study marks the initial identification of HSH2D upregulation in CRC and its role in advancing the progression and chemoresistance of CRC by modulating YAP1. HSH2D emerges as a promising novel target for CRC treatment, suggesting a potential therapeutic strategy. Future research should delve deeper into the specific interaction mechanisms between HSH2D and YAP1, aiming to effectively target this pathway for enhanced CRC treatment.