Xin Zhao

Background: Gastric cancer (GC) remains a major cause of cancer-related mortality worldwide, and Helicobacter pylori (H. pylori) infection is a well-established carcinogenic driver. Cuproplasia, a recently proposed copper-dependent biological state characterized by hyperplasia, metaplasia, and neoplasia, has emerged as a potential mechanistic link between metabolic reprogramming and tumor progression. However, the contribution of cuproplasia-related genes to H. pylori-associated gastric carcinogenesis remains poorly defined. This study aimed to identify cuproplasia-associated genes linked to H. pylori-positive GC and to clarify the functional role and underlying mechanism of LOXL2 in H. pylori-associated gastric cancer progression. Methods: Transcriptomic and clinical data were analyzed to construct a risk model for H. pylori-positive GC based on cuproplasia-related genes. Model performance was evaluated using receiver operating characteristic (ROC), nomogram, decision curve, and survival analyses. LOXL2 was subsequently selected for validation in clinical specimens by immunohistochemistry (IHC), western blotting, and reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Functional assays, including colony formation, wound healing, Transwell invasion, and mechanistic western blotting, were used to determine the biological effects of LOXL2 and H. pylori stimulation in GC cells. Results: A five-gene cuproplasia-related signature showed strong predictive performance for H. pylori-positive GC. Among the candidate genes, LOXL2 emerged as the most prominent biomarker. LOXL2 was significantly overexpressed in H. pylori-infected GC tissues and was associated with tumor size, differentiation status, tumor-node-metastasis (TNM) stage, lymph node metastasis, distant metastasis, and H. pylori positivity. In vitro, LOXL2 overexpression enhanced GC cell proliferation, migration, and invasion, whereas LOXL2 knockdown exerted the opposite effects. H. pylori stimulation further amplified the tumor-promoting effects of LOXL2. Mechanistically, LOXL2 and H. pylori cooperatively increased FAK and Src phosphorylation, upregulated Snail, and suppressed E-cadherin expression, thereby facilitating epithelial-mesenchymal transition (EMT). Conclusions: Rather than serving merely as a prognostic biomarker, LOXL2 functions as a cuproplasia-associated effector that cooperates with H. pylori infection to promote GC progression through activation of the FAK/Src signaling pathway, thereby providing new insights into the molecular mechanisms underlying H. pylori-associated gastric carcinogenesis.

Introduction: Under chronic infections or in tumors, persistent antigen exposure drives CD8 ⁺ T cell exhaustion, a heterogeneous state encompassing a differentiation continuum from stem-like progenitor (Tpex) cells through transitory effector-like (Tex-int) cells to terminally exhausted (Tex-term) subsets. Among these T cell subsets, Tex-int cells serve as the primary population responsible for direct tumor cell killing. However, the intrinsic regulatory mechanisms that govern the Tpex-to-Tex-int transition remain incompletely defined. Methods: In this study, we explore the role of special AT-rich sequence-binding protein 1 (SATB1) in the differentiation of Tex-int cells from their precursors. We observed downregulation of SATB1 during Tpex-to-Tex-int differentiation in tumors. Notably, the genetic ablation of Satb1 in T cells markedly expanded the population of tumor-infiltrating CD8 ⁺ T cells (CD8 ⁺ TILs). Results: Ablating Satb1 not only promoted the differentiation of Tex-int cells from Tpex cells within the tumor microenvironment but also remodeled T cell differentiation in tumor-draining lymph nodes (TdLNs) by expanding the Tpex pool from tumor-specific memory CD8 ⁺ T cells (T _TSM ) and driving the Tpex1 to Tpex2 transition, thereby augmenting Tex-int production in tumors. Although early-stage Tex-int cells in Satb1 -deficient mice displayed transient functional impairment relative to controls, this difference was no longer evident in late-stage tumors, where sustained Tex-int accumulation correlated with significantly suppressed tumor growth and prolonged survival. Discussion: Our results identify SATB1 as a pivotal regulator of exhausted CD8 ⁺ T cell subset differentiation and suggest its targeting as a promising strategy to expand the Tex-int population for enhanced cancer immunotherapy.

Th9/Tc9 cells polarized with TGF-β and IL-4 exhibit superior antitumor efficacy, prompting extensive efforts to optimize their differentiation protocols and augment therapeutic potency. In the current study, we identified IL-36γ as a potent cytokine that synergizes with TGF-β to robustly drive Th9 differentiation both in the presence and absence of IL-4. IL-36γ-programmed Th9 cell subsets exhibited phenotypic and transcriptional profiles identical to classic Th9 cells. Mechanistically, IL-36γ drove Th9 cell differentiation through amplifying key signaling pathways such as STAT6, STAT5, and NF-κB that are essential for classic Th9 programming. Notably, we uncovered a novel regulatory axis wherein IL-36γ upregulates the transactivation factor IκBζ, which directly governs Th9 lineage specification. In in adoptive cell therapy (ACT) models, IL-36γ-polarized Th9 cell subsets demonstrated enhanced antitumor efficacy, attributable to their sustained persistence, reduced exhaustion markers and stem-like/memory properties. Collectively, this study elucidates a previously unrecognized IκBζ-dependent mechanism underpinning Th9 differentiation and highlights the translational potential of IL-36γ-engineered Th9 cells as a valuable ACT strategy for refractory tumors.

The incidence and mortality rates of colorectal cancer (CRC) continue to increase, underscoring the need to identify key gene networks and clinically relev

Objective: To explore the potential involvement of PANoptosis-related genes in gastric cancer susceptibility through multiomics analyses. Methods: Summary-data–based Mendelian randomization was performed by integrating blood-derived methylation, gene expression, and protein quantitative trait loci data with genome-wide association study results. The findings were further evaluated in The Cancer Genome Atlas cohort, followed by protein–protein interaction analysis, drug prediction, and molecular docking. Results: Summary-data–based Mendelian randomization and colocalization analyses identified several traits suggestively associated with gastric cancer risk. Genetically predicted higher expression of apoptosis and caspase activation inhibitor ( AVEN ) and hepatocyte growth factor ( HGF ) as well as higher HGF protein levels were associated with increased risk, whereas higher levels of protein phosphatase 2 regulatory subunit B beta (PPP2R2B) appeared to be protective. Multiomics integration suggested epigenetic regulation of HGF and PPP2R2B . The Cancer Genome Atlas analysis corroborated the dysregulation of these candidates, with high AVEN expression associated with poorer survival. Protein–protein interaction and drug prediction analyses highlighted functional networks and potential therapeutics, supported by molecular docking demonstrating strong HGF - binding affinities. However, these associations did not reach statistical significance in the independent validation cohort, possibly due to limited statistical power. Conclusions: This study identified AVEN , HGF , and PPP2R2B as potential candidate genes for gastric cancer. These findings require further validation in larger cohorts.

Background In order to better understand the trends and hot spots in angiogenesis research in metastatic colorectal cancer, this study uses bibliometric analysis to conduct a systematic analysis. Methods Publications were retrieved from Web of Science Core Collection from 2000 to 2023. All studies were output for further analysis in R and CiteSpace software. Results A total of 5875 studies were recruited via the Web of Science Core Collection from 2000 to 2023 regarding the field of angiogenesis in metastatic colorectal cancer. China and USA were the leading country with top country publications. Among all contributed authors, the main three subjects were determined as cell biology, oncology, and peripheral vascular disease. Furthermore, network analysis of all recruited studies identified 16 research subjects. Next, the top 25 countries, authors references, institutions, and keywords with the strongest citation bursts were identified. To further delineate the growth and change of two types of keywords, Keywords Plus^® and author keywords, a heatmap analysis was employed. Last, the most influential references relating to this field were identified using the strongest citation bursts. Conclusions This study explored the research hot spots and trends of angiogenesis in metastatic colorectal cancer through bibliometric analysis. It would help researchers better understand the current status and future development hotspots of global research to promote further development in this field.

Introduction: Recent findings reveal that the precursors of exhausted CD8⁺ T (CD8⁺ Tpex) cells possess stem-like signatures in tumor immunity, which originate from tumor draining lymph node (TdLN)-derived tumor-specific memory (CD8⁺ T_TSM) cells. Both of these T subsets can be collectively referred to as stem-like CD8⁺ T cells, which demonstrate robust self-renewal ability and can proliferate and differentiate into transitory effector-like exhausted T cells (Tex^int). There are reports that chemotherapeutic drugs can promote the antitumor immune responses of patients by increasing the number of CD8⁺ T cells; however, whether chemotherapeutic drugs increase these two stem-like CD8⁺ T cells remain further exploration.Methods: Tpex cell-associated subpopulations in human colorectal tumors were analyzed by using single-cell sequencing data. CT26 and B16 tumor models of wild type and Eomes conditional knockout mice were constructed, and the changes of T_TSM, Tpex and Tex subsets in mice were dissected by flow cytometry after treatment with decitabine (DAC), doxorubicin (DOX) and 5-Fluorouracil (5-FU).Results: In this study, we demonstrated that DAC and 5-FU expanded CD8⁺ T_TSM cells in TdLNs. At the same time, we validated that DAC and 5-FU substantially promoted the expansion of CD62L⁺CD8⁺ Tpex cells and subsequently increased effector function of CX3CR1⁺ CD8⁺ Tex^int cells. In addition, the conditional knockout of transcription factor Eomes in CD8⁺ T cells partially eliminated DAC-amplified CD62L⁺ CD8⁺ Tpex cells, but had no effect on such CD8⁺ T subset expanded by 5-FU.Conclusion: The present study demonstrated that both DAC and 5-FU promoted the differentiation of stem-like CD8⁺ T_TSM cells in TdLNs and significantly enhanced the differentiation and expansion of stem-like CD62L⁺ CD8⁺ Tpex and CX3CR1⁺ Tex^int cells in tumor microenvironment. The knockout of Eomes partially influenced the role of DAC in promoting the differentiation and expansion of stem-like CD8⁺ T cells.

Background: B7-H3 (or CD276) represents an important costimulatory molecule expressed in many malignant solid tumors, including colorectal cancer (CRC). The receptor of B7-H3 is not known, and the intracellular function of B7-H3 remains obscure. Herein, we report that B7-H3 upregulated the epidermal growth factor heparin-binding epidermal growth factor (HB-EGF), likely by regulating hypoxia-inducible factor 1α (HIF-1α) and thereby promoting the progression of CRC. Methods: Lentiviral transfection was performed on CRC cells to establish stable low-B7-H3 expression cells. A mechanistic analysis with an Agilent human gene expression profiling chip was conducted on them. Clinical data and specimens were collected to detect the connection between B7-H3 and HB-EGF in CRC. Quantitative real-time polymerase chain reaction (qRT-PCR) was conducted to detect the messenger RNA (mRNA) level of B7-H3, HB-EGF, and HIF-1α. Chromatin immunoprecipitation (ChIP) quantitative real-time PCR was conducted. The protein level of HIF-1α and the phosphatidylinositide 3-kinases (PI3K)-protein kinase B (AKT) pathway were detected by western blot. HIF-1α was recovered by lentiviral transfection, and the HB-EGF mRNA levels, proliferation, invasion, and angiogenesis ability were detected. Results: B7-H3 promoted tumor progression through HB-EGF and the PI3K-AKT pathway. As B7-H3 was downregulated, HB-EGF levels were significantly reduced simultaneously, a growth trend that was shown by both CRC cell lines and cancer tissues. In addition, B7-H3 and HB-EGF had significant associations with tumor-node-metastasis (TNM) stage and lymph node metastasis in 50 CRC patients. The binding ability of HIF-1α to the HB-EGF promoter region was significantly decreased in the shB7-H3 RKO group. Western blot revealed that PI3K, AKT, and mammalian target of rapamycin (mTOR) protein amounts and p-AKT and p-mTOR phosphorylation were also downregulated in shB7-H3 RKO cells, suggesting that B7-H3 may regulate HIF-1α via PI3K-AKT signaling. After recovery of the HIF-1α level by lentiviral transfection, the HB-EGF mRNA levels, proliferation, invasion, and angiogenesis in CRC cells recovered as well. Conclusions: B7-H3 may transmit intracellular signals through PI3K-AKT-mTOR-HIF-1α signaling, upregulating HB-EGF. As the final transcription factor of the pathway, HIF-1α regulates the transcription of the HB-EGF gene, thereby promoting HB-EGF expression, which eventually mediates cell proliferation, invasion, and angiogenesis and promotes the progression of CRC.

Pancreatic cancer (PC), the third leading cause of cancer-related death in the U.S., is frequently found too late to be cured by traditional chemotherapy. Expression of B7 homolog 6 (B7H6), a member of the B7 family of immunoreceptors, has been found in PC and several other cancers. B7H6 is a ligand for cytotoxicity triggering receptor 3 (NKp30), which is expressed on NK cells. Here, we demonstrate that B7H6 can be detected in PC tissues but not normal organs. Its expression in patients associated significantly with tumor differentiation grade and lymphatic metastasis. The soluble form of B7H6 was detected in the PC patients’ sera, and its concentration associated with tumor differentiation grade and tumor, node, metastasis (TNM) stages. Also, higher levels of B7H6 in PC patients’ malignant tissues or serum correlated with shorter overall survival. In vitro, downregulation of B7H6 by CRISPR/Cas9 or siRNA technology had no significant impact on the viability or mobility of PC cells. Instead, knocking out B7H6 sensitized PC cells to NK-mediated cytotoxicity and cytokine production. These results indicate that B7H6 not only serves as a negative prognostic marker but also acts as an immune modulator in PC.

Background: Colon cancer is a common malignant tumor of the digestive tract, and its incidence is ranked third among gastrointestinal tumors. The present study aims to investigate the role of a novel circular RNA (circCSPP1) in colon cancer and its underlying molecular mechanisms. Methods: Bioinformatics analysis and reverse transcription-quantitative PCR were used to detect the expression levels of circCSPP1 in colon cancer tissues and cell lines. The effects of circCSPP1 on the behavior of colon cancer cells were investigated using CCK-8, transwell and clonogenic assays. Bioinformatics analysis along with luciferase, fluorescence in situ hybridization and RNA pull-down assays were used to reveal the interaction between circCSPP1, microRNA (miR)-431, Rho associated coiled-coil containing protein kinase 1 (ROCK1) and zinc finger E-box binding homeobox 1 (ZEB1). Results: It was found that circCSPP1 expression was significantly upregulated in colon cancer tissues and cell lines. Overexpression of circCSPP1 significantly promoted the proliferation, migration and invasion of colon cancer cells, whereas silencing of circCSPP1 exerted opposite effects. Mechanistically, circCSPP1 was found to bind with miR-431. In addition, ROCK1 and ZEB1 were identified as the target genes of miR-431. Rescue experiments further confirmed the interaction between circCSPP1, miR-431, ROCK1 and ZEB1. Moreover, circCSPP1 promoted the expression level of ROCK1, cyclin D1, cyclin-dependent kinase 4, ZEB1 and Snail, and lowered the E-cadherin expression level. Conclusion: Taken together, the findings of the present study indicated that circCSPP1 may function as a competing endogenous RNA in the progression of colon cancer by regulating the miR-431/ROCK1 and miR-431/ZEB1 signaling axes.