Objective This study aimed to clarify how cancer-associated fibroblast-derived exosomal circ_0067557 (CAF-exo circ_0067557) promotes epithelial-mesenchymal transition (EMT) in colorectal cancer (CRC) through BHLHE40-mediated transcriptional activation of OTUB2.Methods CAF-derived exosomes were isolated and characterized from CRC tissues, while normal fibroblast-derived exosomes were obtained from adjacent normal tissues. The transfer of circ_0067557 was tracked using fluorescence labeling and endocytosis assays. Expression of circ_0067557, BHLHE40, OTUB2, and EMT-related markers was assessed by qRT-PCR, Western blotting, and immunofluorescence. Gain- and loss-of-function models were established to determine the biological role of circ_0067557. RNA sequencing identified OTUB2 as a key downstream target. ChIP-qPCR and dual-luciferase reporter assays were performed to verify BHLHE40 binding to the OTUB2 promoter. RIP and RNA pull-down assays confirmed the interaction between circ_0067557 and BHLHE40. Mouse xenograft and tail vein metastasis models were used to evaluate the oncogenic effects of circ_0067557 in vivo.Results CAF-exos successfully delivered circ_0067557 into CRC cells, significantly enhancing their proliferative, migratory, invasive, and EMT capabilities. Mechanistically, circ_0067557 interacted with and recruited BHLHE40, thereby increasing its binding to the OTUB2 promoter and promoting OTUB2 transcription. In vivo experiments further confirmed that circ_0067557 markedly enhanced tumor growth and distant metastasis through the BHLHE40/OTUB2 signaling axis.Conclusion CAF-exo circ_0067557 promotes EMT, invasion, metastasis, and tumor progression in CRC by recruiting BHLHE40 and activating OTUB2 transcription. These findings reveal a novel mechanism by which the tumor microenvironment (TME) regulates cancer cell metastasis.

93 Background: Neoadjuvant chemoradiotherapy (nCRT) has brought out a higher rate of complete response and anal sphincter preservation for locally advanced rectal cancer (LARC). Combining immunotherapy with radiotherapy and chemotherapy has shown synergistic effects. This study a evaluates the efficacy and safety of long-term nCRT with camrelizumab (a PD-1 inhibitor) for LARC and low rectal cancer requiring anal preservation. Methods: In this single-arm, prospective, phase II trial, patients with newly diagnosed LARC (cT3-4N0M0 or cT1-4N+M0) and low rectal cancer requiring sphincter preservation (cT2N0M0, <5 cm from anal verge) were enrolled. All patients received long-course radiotherapy (50Gy/45Gy/25f) with concurrent chemotherapy [T2-3N0M0: capecitabine (1650 mg/m 2 /d, bid, d1-14, Q3W); T4N0M0 or T1-4N+M0: capecitabine + oxaliplatin (130 mg/m 2 , d1, Q3W)], plus camrelizumab (200 mg, d1, Q3W) for 4 cycles. Researchers evaluated patients after neoadjuvant therapy, followed by surgical resection. The primary endpoint was pathological complete response rate (pCR), and secondary endpoints included clinical complete response rate (cCR), objective response rate (ORR), disease control rate (DCR), disease-free survival, overall survival and safety. Results: Between January 2024 and December 2025, 58 patients were enrolled with 56 completing the efficacy evaluation. Among them, 2 patients achieved cCR, 22 patients had partial response, and 32 stable disease. The ORR was 42.9%, and DCR was 100%. Of 58 patients, 43 underwent surgery, 4 adopted wait and watch strategy (2 achieved cCR and 2 achieved near-cCR), 7 continued observation without surgery, 2 declined surgery, and 2 were in going therapy. All surgical patients achieved R0 resection, with pCR rate at 46.5% (20/43). Among the16 patients with low rectal cancer, 11 (68.8%) achieved anal sphincter preservation. In tumor regression grade (Mandard five-grade standard), 20 patients were Grade 1, 16 with Grade 2, 5 with Grade 3, and 2 with Grade 4. Regarding safety, 56 patients (100%) experienced treatment-related adverse effects, including lymphopenia (89.3%), reactive capillary endothelial proliferation (67.9%), leukopenia (39.3%), thrombocytopenia (33.9%), diarrhea (21.4%), fatigue (14.3%), and nausea (8.9%). Four patients reported Grade 3 adverse effects, predominantly lymphopenia (3.6%) and thrombocytopenia (3.6%). Patients generally tolerated treatment well, with symptoms managed and relieved symptomatically. Conclusions: Long-term neoadjuvant chemoradiotherapy (nCRT) with camrelizumab has shown promising rates of pathological complete response and clinical safety in patients with locally advanced and low rectal cancer aiming to preserve anal function. This ongoing study anticipates final results that may further enhance survival outcomes for these patients. Clinical trial information: NCT06304545 .

Macrophage have shown great promise in the clinical treatment for head and neck cancer (HNC). However, macrophage therapies confront significant hurdles due to a deficiency of tumor-specific targets and tumor resistance. Herein, we report the development of a magnetotactic bacterial-reprogrammed T-cell receptor macrophage (TCR-M) that integrates human papillomavirus (HPV) associated HNC specific targeting with innate macrophage infiltration capacity, enabling potent immune cytotoxicity against head and neck squamous cell carcinoma (HNSCC). By internalizing magnetotactic bacteria AMB-1, the TCR-M combination achieves tumor-specific targeting and controllable magnetic responsiveness to elicit durable anti-tumor photoimmune response. Notably, AMB-1 incorporation endows macrophages with magnetic navigation capability and laser-responsive polarization into Bac@TCR-M + L, which acquires M1 phenotypes with sustained anti-tumor photoimmune response. Under magnetic actuation, Bac@TCR-M + L demonstrates significantly enhanced tumor site accumulation compared to untreated TCR-M, accompanied by elevated secretion of TNF-α and IFN-γ and augmented reactive oxygen species (ROS) production. Furthermore, Bac@TCR-M + L re-educates tumor-associated macrophages (TAMs) into anti-tumor M1 phenotypes, achieving collaborative tumor destruction. Our findings establish Bac@TCR-M + L as a paradigm-shifting macrophage therapy that integrates microbial engineering, robotic control, and cell therapy, yielding durable anti-tumor effects in HNC treatment and opening new avenues for overcoming cancer resistance.

BackgroundSemaphorins (SEMAs), originally identified as axon guidance factors, have been found to play crucial roles in tumor growth, invasiveness, neoangiogenesis, and the modulation of immune responses. However, the prognostic value of SEMA-related genes in colorectal cancer (CRC) remains unclear.MethodsWe applied a novel machine learning framework that incorporated 10 machine learning algorithms and their 101 combinations to construct a SEMAs-related score (SRS). Multi-omics analysis was performed, including single-cell RNA sequencing (scRNA-seq), and spatial transcriptome (ST) to gain a more comprehensive understanding of the SRS. A series of cell experiments were conducted to prove the impact of key genes on CRC biological behavior.ResultA consensus SRS was finally constructed based on a 101-combination machine learning computational framework, demonstrating outstanding performance in predicting overall survival. Moreover, distinct biological functions, mutation burden, immune cell infiltration, and immunotherapy response were observed between the high- and low-SRS groups. scRNA-seq and ST demonstrated unique cellular heterogeneity in CRC. We observed that SRS-high and SRS-low malignant epithelial cells exhibit different biological characteristics. High SRS malignant epithelial cells interact with myeloid and endothelial cells via SPP1 and COL4A2-ITGAV-ITGB8 pathways, respectively. Low SRS cells engage with myeloid and endothelial cells through MIF and JAG1-NOTCH4 pathways. Additionally, knocking down SEMA4C significantly inhibits the proliferation and invasion of CRC cells, while promoting apoptosis in vitro.ConclusionSRS could serve as an effective tool to predict survival and identify potential patients benefiting from immunotherapy in CRC. It also reveals tumor heterogeneity and provides valuable biological insights in CRC.

Background Cancer associated fibroblasts (CAFs) can remodel tumor microenvironment by secreting exosomes. This study aimed to investigate the role of exosomes derived from cancer-associated fibroblasts in colorectal cancer (CRC) progression. Methods Circular RNA (circRNA) array was used to identify differentially expressed circRNAs in exosomes from normal fibroblasts (NFs) and CAFs, and confirmed one differentially expressed circRNA circ_0067557 by real-time PCR. The effect of circ_0067557 on proliferation, metastasis, chemoresistance and apoptosis was verified by wound heal, tranwell, CCK8, sphere-forming and flow cytometry assay. Results Circ_0067557 expression in exosomes from CAFs was higher than those from NFs. CAF-derived exosomes promoted the proliferation, migration, invasion and chemoresistance of CRC cells while suppressed apoptosis. Silencing of circ_0067557 inhibited malignant phenotypes of CRC cells by targeting Lin28A and Lin28B. Moreover, CAF-derived exosomes enhanced the growth of CRC xenograft tumors. Conclusion Circ_0067557/Lin28A and Lin28B signal axis may be a potential therapy target for CRC.

G-quadruplexes (G4s) can recruit transcription factors to activate gene expression, but detailed mechanisms remain enigmatic. Here, we demonstrate that G4s in the CCND1 promoter propel the motility in MAZ phase-separated condensates and subsequently activate CCND1 transcription. Zinc finger (ZF) 2 of MAZ is a responsible for G4 binding, while ZF3-5, but not a highly disordered region, is critical for MAZ condensation. MAZ nuclear puncta overlaps with signals of G4s and various coactivators including BRD4, MED1, CDK9 and active RNA polymerase II, as well as gene activation histone markers. MAZ mutants lacking either G4 binding or phase separation ability did not form nuclear puncta, and showed deficiencies in promoting hepatocellular carcinoma cell proliferation and xenograft tumor formation. Overall, we unveiled that G4s recruit MAZ to the CCND1 promoter and facilitate the motility in MAZ condensates that compartmentalize coactivators to activate CCND1 expression and subsequently exacerbate hepatocarcinogenesis.

[Objective] ZEB2 is known to be participated in several cancers, yet there is no comprehensive analysis about its function in pan-cancer. This study aimed to investigate the role ZEB2 plays in pan-cancer. [Methods] The data was downloaded from the University of California Santa Cruz (UCSC) Xena and the Cancer Genome Atlas (TCGA). The mRNA expression status of ZEB2 was studied in the TCGA_GTEx samples, TCGA samples and paired samples in TCGA, respectively, and protein expression status was obtained from the University of Alabama at Birmingham Cancer data analysis Portal (UALCAN). Kaplan-Meier analysis was applied to 33 kinds of tumors in TCGA, then among the cancers that ZEB2 can affect prognosis, clinical correlation analysis and univariate and multivariate Cox regression analysis were performed. Furthermore, to confirm the prognostic value of ZEB2 in cancers, nomogram models were constructed on lung adenocarcinoma (LUAD) and stomach adenocarcinoma (STAD). The relationship between ZEB2 mRNA expression and immune cell infiltrates was determined by tumor immune estimation resource (TIMER2.0). Protein-protein interaction (PPI) networks were constructed by the STRING. Functional enrichment analysis was also performed using the “ClusterProfiler” package. [Results] ZEB2 was expressed in most tumors, either upregulated or downregulated, and correlated with clinical characteristics in a part of tumors. The expression level of ZEB2 was also found to impact the prognosis of tumors. Additionally, ZEB2 expression was significantly related to immune cells infiltration level in tumor microenvironment. [Conclusion] we propose that ZEB2 has the potential to be a prognostic biomarker and a promising target for cancer immunotherapy.

Introduction Gastric cancer (GC) is the fifth most prevalent cancer globally, with the third highest case fatality rate. Neutrophil extracellular traps (NETs) are a reticulated structure of DNA, histones, and antimicrobial peptides produced by active neutrophils that trap pathogens. Even though NETs are associated with poorer recurrence-free survival (RFS) and overall survival (OS), the specifics of this interaction between NETs and cancer cells are yet unknown. Methods The keywords “neutrophil extracellular traps and gastric cancer” were used in the GEO database for retrieval, and the GSE188741 dataset was selected to obtain the NETs-related gene. 27 NETs-related genes were screened by univariate Cox regression analysis ( p < 0.05). 27 NETs-related genes were employed to identify and categorize NETs-subgroups of GC patients under the Consensus clustering analysis. 808 GC patients in TCGA-STAD combined with GES84437 were randomly divided into a training group ( n = 403) and a test group ( n = 403) at a ratio of 1:1 to validate the NETs-related signature. Results Based on Multivariate Cox regression and LASSO regression analysis to develop a NETs-related prognosis model. We developed a very specific nomogram to improve the NETs-clinical score’s usefulness. Similarly, we also performed a great result in pan-cancer study with NETs-score. Low NETs scores were linked to higher MSI-H (microsatellite instability-high), mutation load, and immune activity. The cancer stem cell (CSC) index and chemotherapeutic treatment sensitivity were also connected to the NET score. Our comprehensive analysis of NETs in GC suggests that NETs have a role in the tumor microenvironment, clinicopathological features, and prognosis. Discussion The NETs-score risk model provides a basis for better prognosis and therapy outcomes in GC patients.

The occurrence and progression of tumors can be established through a complex interplay among tumor cells undergoing epithelial-mesenchymal transition (EMT), invasive factors and immune cells. In this study, we employed single-cell RNA sequencing (scRNA-seq) and spatially resolved transcriptomics (ST) to evaluate the pseudotime trajectory and spatial interactive relationship between EMT-invasive malignant tumors and immune cells in primary colorectal cancer (CRC) tissues at different stages (stage I/II and stage III with tumor deposit). Our research characterized the spatiotemporal relationship among different invasive tumor programs by constructing pseudotime endpoint-EMT-invasion tumor programs (EMTPs) located at the edge of ST, utilizing evolution trajectory analysis integrated with EMT-invasion genes. Strikingly, the invasive and expansive process of tumors undergoes remarkable spatial reprogramming of regulatory and immunosuppressive cells, such as myeloid-derived suppressor cells (MDSCs), tumor-associated macrophages (TAMs), regulatory T cells (Treg), and exhausted T cells (Tex). These EMTP-adjacent cell are linked to EMT-related invasion genes, especially the C-X-C motif ligand 1 (CXCL1) and CXCL8 genes that are important for CRC prognosis. Interestingly, the EMTPs in stage I mainly produce an inflammatory margin invasive niche, while the EMTPs in stage III tissues likely produce a hypoxic pre-invasive niche. Our data demonstrate the crucial role of regulatory and immunosuppressive cells in tumor formation and progression of CRC. This study provides a framework to delineate the spatiotemporal invasive niche in CRC samples.

Background: Tumor cell epithelial-mesenchymal transition (EMT) is crucial for cancer progression. Molecular profiling of single cells has enhanced our understanding of the molecular basis of EMT but losing the crucial spatial information of invasion process in tumor microenvironment.Methods: This study utilized single cell RNA sequencing and spatially resolved transcriptomic (SRT) to assess rectum carcinoma tissues (stage I vs. stage III with tumor deposit). To evaluate the grow direction of malignant programs, we proposed up-regulated and down-regulated invasion scores by associating with pseudo-time. Cell-cell interaction and relocation the tumor programs with EMT-status in SRT are also performed.Findings: The research characterizes the spatio-temporal relationship for different stage of tumor programs by constructing an EMT-time-dependent invasive signature. The research also revealed the spatial and physiologic relationship between advanced tumor programs with highly EMT status. Adjacent immune cells in TME are also have important interaction with the EMT tumor program, including cytotoxic CD8+ T cells, neutrophils, and terminally exhausted T cells, with interaction of MIF- (S100A4/SREBF1/ERRFI1), TGFB1-JAG1, and EGFR-ADAM17.Interpretation: The current study provides a framework for studying the underline mechanisms of tumor cell invasion across EMT in complex tumor progression.Funding: This study was funded by the National Natural Science Foundation of China (U20A20376); the Free Exploration Foundation of Science, 45 Technology, and Innovation Commission of Shenzhen Municipality (No. JCYJ2018307151238174); and the Science and Technology Planning Project of Guangdong Province (No. 2017A020215014).Declaration of Interest: We declare there are no any competing financial interests in relation to this work.Ethical Approval: This study was approved by the Ethics Committee of Harbin Medical University Cancer Hospital. Both study patients provided written informed consent before tissue collection for this study.

Abstract Exosomes derived from cancer-associated fibroblasts (CAFs) may be crucial in colorectal cancer (CRC) progression, although the underlying function and mechanism of CAF-derived exosomes in CRC are still unclear. We used a circular RNA (circRNA) array to identify differentially expressed circRNAs in exosomes from normal fibroblasts (NFs) and CAFs. One differentially expressed circRNA was circ_0067557, whose expression was verified by RT-qPCR. The target genes of circ_0067557 (Lin28A and Lin28B) were identified by RNA-pull down, RNA-IP, and RNA-EMSA. The biological behavior of CRC cells and CRC xenograft tumors were examined by multiple assays. Circ_0067557 expression in exosomes from CAFs was higher than in exosomes from NFs. Functionally, CAF-derived exosomes could induce proliferation, migration, invasion, chemoresistance, EMT, and the cancer stem cell (CSC) features of CRC cells while suppressing apoptosis. In addition, we confirmed that circ_0067557 could bind to its target genes (Lin28A and Lin28B). Silencing of circ_0067557 prevented the malignant phenotype of the CRC cell by targeting Lin28A and Lin28B. Moreover, CAF-derived exosomes enhanced the growth of CRC xenograft tumors, suppressed apoptosis, and upregulated Lin28A and Lin28B through circ_0067557. Circ_0067557/Lin28A and Lin28B signal axis may be a potential therapy target for CRC patients.

Ephrin Type-A Receptor 3 (EphA3) and Ephrin Type-B Receptor 6 (EphB6) belong to the ephrin receptor group consisting of the largest subset of receptor tyrosine kinases (RTKs) and are essential for neurogenesis and embryogenesis.The current study aimed to evaluate their functional roles in transforming colorectal epithelial cells and dissect the underlying molecular mechanisms.We observed altered EphA3 and EphB6 expression in tumor tissues as compared to normal tissues in a tissue microarray study.Enforced EphB6 expression promoted IMCE cell proliferation, migration, and invasion in vitro and tumor formation in nude mice, with a stronger oncogenic activity than EphA3.Pathway analysis of differentially expressed genes from a gene microarray study provided important insight into potential mechanisms through which EphB6 may regulate the malignant transformation of colorectal epithelial cells.This study represents the first demonstration of EphB6 in enhancing colorectal epithelial cell transformation, suggesting its stipulative role in the early stage of colorectal tumorigenesis.Our findings primarily uncover novel biomarkers and therapeutic targets of colorectal cancer.

Accumulating evidence suggested the participation of long noncoding RNAs (lncRNAs) in regulating various biological processes so as to affecting cancer progression. However, the functional role of most lncRNAs in colorectal carcer (CRC) is still largely covered. In the present study, we disclosed SNHG14 as a carcinogene in CRC development, as it was low-expressed in normal colon tissues but markedly upregulated in CRC cell lines. Besides, SNHG14 contributed to CRC cell proliferation, motility and EMT in vitro, and inhibition of it confined CRC tumor growth and liver metastasis in vivo. Next, the mechanistic investigations confirmed that SNHG14-promoted CRC progression was mediated by EPHA7, which was negatively regulated by SNHG14 in CRC via an EZH2-dependent way. Importantly, EZH2 was proved as a transcription factor of EPHA7 and functioned as a repressor in EPHA7 transcription by enhancing methylation on EPHA7 promoter. Meanwhile, SNHG14 increased EZH2 expression in CRC via stabilizing its mRNA by interacting with FUS, and via freeing its mRNA from miR-186-5p-induced silence. All in all, our observations demonstrated that SNHG14 serves as a facilitator in CRC through targeting EZH2-repressed EPHA7 by enhancing EZH2 via recruiting FUS and absorbing miR-186-5p, indicating a promising new road for CRC diagnosis and treatment.

BACKGROUND:With more than 600,000 mortalities each year, colorectal cancer (CRC) is the third most commonly diagnosed type of cancer worldwide. Recently, mechanisms involving noncoding RNAs have been implicated in the development of CRC.METHODS:We examined expression levels of lncRNA CRNDE and miR-181a-5p in 64 cases of CRC tissues and cell lines by qRT-PCR. Gain-of-function and loss-of-function assays were performed to examine the effect of CRNDE and miR-181a-5p on proliferation and chemoresistance of CRC cells. Using fluorescence reporter and western blot assays, we also explored the possible mechanisms of CRNDE in CRC cells.RESULTS:In this study, we found that the expression levels of the CRNDE were upregulated in CRC clinical tissue samples. We identified microRNA miR-181a-5p as an inhibitory target of CRNDE. Both CRNDE knockdown and miR-181a-5p overexpression in CRC cell lines led to inhibited cell proliferation and reduced chemoresistance. We also determined that β-catenin and TCF4 were inhibitory targets of miR-181a-5p, and that Wnt/β-catenin signaling was inhibited by both CRNDE knockdown and miR-181a-5p overexpression. Significantly, we found that the repression of cell proliferation, the reduction of chemoresistance, and the inhibition of Wnt/β-catenin signaling induced by CRNDE knockdown would require the increased expression of miR-181a-5p.CONCLUSIONS:Our study demonstrated that the lncRNA CRNDE could regulate the progression and chemoresistance of CRC via modulating the expression levels of miR-181a-5p and the activity of Wnt/β-catenin signaling.

Altered mean platelet volume (MPV) is implicated in several malignancies. However, the clinicopathological significance and prognostic value of MPV in colorectal cancer (CRC) is still elusive. The purpose of this study was to elucidate the predictive significance of MPV in CRC. The retrospective study recruited 509 consecutive CRC patients between January 2009 and December 2009. The relationships between MPV and clinicopathological characteristics were analyzed. Kaplan-Meier method and Cox regression were used to evaluate the prognostic impact of MPV. Of the 509 CRC patients, high MPV levels were detected in 150 (29.5%) patients. Elevated MPV was associated with tumor differentiation (p < 0.001). Patients with increased MPV had poor overall survival compared with those with normal level (60.0% vs. 83.6%, log-rank test, p = 0.035). Cox regression analysis showed that MPV was an independent prognostic factor in CRC (HR = 1.452, 95% CI = 1.118–1.884, p = 0.005). In conclusion, MPV is easily available in routine blood test. Elevated MPV might act as a marker of prognosis and therapeutic target for CRC.

Ephrin Type-A Receptor 3 (EphA3) belongs to the ephrin receptor subfamily of the protein tyrosine kinase family, and plays an important role in embryogenesis and neurogenesis. This study aimed to investigate the role of EphA3 in promoting malignant transformation of colorectal epithelial cells, and explore underlying molecular mechanisms. Colorectal cancer tissue specimens from 68 patients were analyzed for EphA3 expression. EphA3 expression levels were manipulated in rat colon epithelial cell lines. We found that EphA3 expression level in tumor tissues was associated with patient age (P = 0.015), tumor differentiation (P = 0.001), and lymph node metastasis (P = 0.039). Overexpression of EphA3 and its constitutively active mutants promoted colony formation, migration and invasion, and tumorigenicity of colon epithelial cells in nude mice. The cDNA and lncRNA microarray profiling data revealed that differentially expressed genes and lncRNAs in EphA3 or mutant-transfected cells were associated with cell proliferation, invasion and angiogenesis. Our findings reveal the mechanisms underlying the oncogenic activities of EphA3 in colorectal cells, which could provide novel targets for the prevention, early diagnosis, and treatment of colorectal cancer.

The erythropoietin-producing hepatocyte (Eph) family tyrosine kinases play important roles in tumorigenesis and cancer aggression. In this study, we investigated the role of EphB6 in oncogenic transformation of colorectal epithelial cells in vitro and in vivo. EphB6 is upregulated in human colorectal cancer (CRC) tissues as compared to normal tissues, and its overexpression promotes proliferation, migration and invasion by IMCE colorectal adenoma cells, in which one Apc allele is mutated. EphB6 overexpression together with Apc mutation leads to the development of colorectal tumors in vivo. Expression microarrays using mRNAs and lncRNAs isolated from EphB6-overexpresssing IMCE and control cells revealed a large number of dysregulated genes involved in cancer-related functions and pathways. The present study is the first to demonstrate that EphB6 overexpression together with Apc gene mutations may enhance proliferation, invasion and metastasis by colorectal epithelial cells. Microarray data and pathway analysis of differentially expressed genes provided insight into possible EphB6-regulated mechanisms promoting tumorigenesis and cancer progression. EphB6 overexpression may represent a novel, effective biomarker predictive of cell proliferation, invasion and metastasis patterns in CRC tumors.