Immunity, Inflammation and Disease

Tumor Cell‐Expressed Herpesvirus Entry Mediator Regulates Proliferation and Adaptive Immunity in Ovarian Cancer

ABSTRACT Background Ovarian cancer (OvCa) is a prevalent gynecological malignancy with an increasing incidence and high mortality rate. Although the role of the herpesvirus entry mediator (HVEM), encoded by the TNFRSF14 gene, is currently considered pivotal in various types of cancer, the regulation of tumor cell‐expressed HVEM in OvCa remains inadequately understood. Methods Specimens were used to detect HVEM expression via quantitative RT‐PCR and flow cytometry. The proliferation of the murine OvCa cell line ID8 was determined using the Cell Counting Kit‐8, colony formation, and EdU staining assays. The immune constituents within the ascites fluid and spleen of tumor‐bearing mice were analyzed by flow cytometry. Bioinformatics analysis was performed to explore cytokines, chemokines, and signaling pathways regulated by HVEM, and differential expression levels were confirmed via quantitative RT‐PCR and western blot analysis. Results Herein, we identified a significant upregulation of HVEM in OvCa tissues compared with that in benign tissues and observed dominant expression of HVEM in CD45⁻EpCAM⁺ subsets in OvCa specimens. Tumor cell‐expressed HVEM was found to promote OvCa cell proliferation by partly activating spliced X‐box‐binding protein 1 (XBP1s)‐c‐Myc signaling. In mouse models, knockdown of Tnfrsf14 in ID8 cells alleviated OvCa progression and specifically affected the frequency and function of T cells in the ascites fluid and spleen. In addition, tumor cell‐expressed HVEM altered chemokine expression (CXCL1/9/10/11 and CCL2/4/5) and STAT signal activation (STAT5 and STAT6) in ID8 cells. Conclusion This study investigated the effects of HVEM on OvCa and validated its potential as a therapeutic marker for treating OvCa.

Sufficiently activated mature natural killer cells derived from peripheral blood mononuclear cells substantially enhance antitumor activity

AbstractBackgroundPeripheral blood‐derived natural killer (NK) cells spontaneously lyse tumor cells without prior sensitization. However, NK cells in peripheral blood (PBNK cells) are in a resting state and exhibit inhibitory phenotypes and impaired cytotoxicity. Thus, strengthening the cytotoxic effector function of PBNK cells and improving NK cell expansion in vitro for a convenient allogeneic therapy are essential.Materials and MethodsPure cytokine activation and expansion of NK cells (super NK [SNK]) from peripheral blood mononuclear cells were studied. Markers of activated and inhibited NK cells and cytokine secretion by NK cells were examined using flow cytometry. NK cell antitumor activity in vitro was assessed using lactate dehydrogenase (LDH) cytotoxicity assay and an Incucyte real‐time imaging system. Additionally, the function of SNK cells against ascites caused by ovarian cancer in NOD‐Prkdc(em26Cd52)il2rg(em26Cd22)/Nju (NCG) mice was determined. In a further investigation of the differences between PBNK and SNK, the mRNA of both cells was sequenced and analyzed.ResultsHuman peripheral blood mononuclear cells showed selective NK cell expansion upon cytokine activation and culture. Both SNK and PBNK cells expressed activation markers, but at different levels, and SNK cells secreted more cytokines related to cytotoxicity than PBNK cells did. Accordingly, SNK cells exhibited strong antitumor activity ex vivo and improved NCG mice survival after intraperitoneal ovarian cancer transplantation. Mechanistically, SNK cells expressed more genes associated with nucleotide metabolism, fatty acid, and ATP metabolism than PBNK cells.ConclusionSNK cells derived from peripheral blood mononuclear cells have sufficiently activated mature characteristics and high antitumor activity, rendering them a highly promising and essential therapeutic approach for cancer treatment.

Exploring the prognostic significance of immunogenic cell death‐related genes as risk biomarkers in cervical cancer

AbstractBackgroundImmunogenic cell death (ICD) is a process in which dying cells stimulate an immune response. It is a regulated form of cell death that can remodel the tumor microenvironment (TME) and activate the immune system, making immunotherapy more effective. This work was designed to identify prognostic gene features associated with ICD in cervical cancer (CC).MethodsBased on CC datasets and a set of ICD‐related genes obtained from public databases, we first filtered out ICD‐related genes unrelated to CC survival using univariate analysis. Subsequently, LASSO regression and multivariate Cox regression analysis were employed to develop prognostic feature genes based on ICD. For the construction and validation of the model, eight genes (CXCL1, IL1B, TNF, YKT6, PDIA3, ROCK1, CXCR3, and CLEC9A) were chosen. A nomogram was created to forecast the prognosis of CC individuals, and Kaplan–Meier curves were utilized to explore the survival disparities among different risk groups of CC individuals.ResultsssGSEA analysis was employed to investigate immune differences between two risk groups, revealing that the low‐risk group exhibited elevated levels of immune cell infiltration, enhanced activation of immune function, and a higher immunophenoscore compared with the other group, which highlighted the relevance of ICD to TME.ConclusionWe constructed a prognostic model based on genetic biomarkers of ICD for prognostic prediction of CC patients. Our model demonstrated excellent discriminative and calibration capabilities, providing a valuable tool for prognostic prediction and assessing the potential efficacy of immunotherapy in CC.

The potential value of cancer‐testis antigens in ovarian cancer: Prognostic markers and targets for immunotherapy

AbstractBackgroundTumor immunotherapy has become an important adjuvant therapy after surgery, radiotherapy, and chemotherapy. In recent years, the role of tumor‐associated antigen (TAA) in tumor immunotherapy has become increasingly prominent. Cancer‐testis antigen (CTA) is a kind of TAA that is highly restricted in a variety of tumors and can induce an immune response.AimsThis review article aimed to evaluate the role of CTA on the progression of ovarian cancer, its diagnostic efficacy, and the potential for immunotherapy.MethodsWe analyzed publications and outlined a comprehensive of overview the regulatory mechanism, immunogenicity, clinical expression significance, tumorigenesis, and application prospects of CTA in ovarian cancer, with a particular focus on recent progress in CTA‐based immunotherapy.ResultsThe expression of CTA affects the occurrence, development, and prognosis of ovarian cancer and is closely related to tumor immunity.ConclusionCTA can be used as a biomarker for the diagnosis and prognosis evaluation of ovarian cancer and is an ideal target for antitumor immunotherapy. These findings provide novel insights on CTA in the improvement of diagnosis and treatment for ovarian cancer. The successes, current challenges and future prospects were also discussed to portray its significant potential.

The role of inflammatory factors and T‐cell subsets in the diagnosis of recurrence in epithelial ovarian cancer patients and the effect of olaparin treatment on them

AbstractBackgroundThe aim of the study is to investigate the role of serum inflammatory factors and T‐cell subsets in the diagnosis of recurrence in epithelial ovarian cancer patients and the effect of olaparib on inflammatory factor and T‐lymphocyte subsets in patients with recurrent epithelial ovarian cancer.MethodsIn this study, 100 patients diagnosed as recurrent epithelial ovarian cancer in our hospital and 100 patients without recurrent epithelial ovarian cancer in the same period were selected. According to the treatment plan, the recurrent patients were divided into conventional therapy group (Paclitaxel and Carboplatin) and combined therapy group (Paclitaxel, Carboplatin, and olaparib). The levels of serum inflammatory factors were evaluated by enzyme‐linked immunosorbent assay. The peripheral blood T‐lymphocyte subsets in each group were detected by flow cytometry.ResultsCompared with nonrecurrent patients, recurrent patients have higher serum interleukin‐6 (IL‐6) and tumor necrosis factor‐α (TNF‐α) levels (p < .05), and lower interferon‐γ (IFN‐γ) level and the CD4+/CD8+ ratio. After adjusting for confounding factors, the results showed that the serum IL‐6, IFN‐γ, and TNF‐α levels were influencing factors of recurrence in epithelial ovarian cancer patients. The area under the receiver operating curve and the sensitivity of serum TNF‐α in predicting ovarian cancer recurrence were higher than those of IL‐6 and IFN‐γ. After secondary chemotherapy and/or olaparib maintenance treatment, the IL‐6 (p < .001) and TNF‐α (p < .001) levels in combined therapy group were lower than those in the conventional therapy, whereas the IFN‐γ level (p < .001), the CD4+ T‐cell proportion (p = .0069) and the CD4+/CD8+ ratio (p = .0201) were higher than those in the conventional therapy.ConclusionThe serum IL‐6, TNF‐α, and IFN‐γ levels were closely related to the recurrence of ovarian cancer. Olaparib maintenance treatment can significantly decrease the IL‐6 and TNF‐α level, and increase IFN‐γ level and the CD4+/CD8+ ratio in patients with recurrent ovarian cancer.

EXPRESSION OF CONCERN: CircZNF124 regulates cell proliferation, leucine uptake, migration and invasion by miR‐199b‐5p/SLC7A5 pathway in endometrial cancer

AbstractBackgroundRecent studies have revealed that circular RNA participates in endometrial carcinoma (EC) progression. Here we investigated the role of circRNA zinc finger protein 124 (circZNF124) in EC genesis and underlying mechanism.MethodsThe expression levels of circZNF124, microRNA‐199b‐5p (miR‐199b‐5p) and solute carrier family 7 member 5 (SLC7A5) were detected by quantitative real‐time polymerase chain reaction. The expression of SLC7A5 and other indicated marker proteins was determined by western blot analysis. For functional assay, cell proliferation, leucine uptake and metastasis were investigated by total cell number, cell counting kit‐8, cell colony formation, leucine uptake or transwell assay. The interaction between miR‐199b‐5p and circZNF124 or SLC7A5 was predicted by starbase online database, and identified by mechanism assays. The impact of circZNF124 absence on tumor growth in vivo was revealed by xenograft mouse model assay. Immunohistochemistry assay was implemented to detect the positive expression rate of nuclear proliferation marker (Ki67).ResultsCircZNF124 and SLC7A5 expression were significantly increased, while miR‐199b‐5p was decreased in EC tissues and cells compared with normal endometrial tissues or cells. CircZNF124 expression was closely associated with EC severity and lymph node metastasis. Additionally, circZNF124 depletion repressed cell proliferation, leucine uptake, migration and invasion in both HEC1A and Ishikawa cells. CircZNF124 regulated SLC7A5 expression by binding to miR‐199b‐5p. MiR‐199b‐5p inhibitors or SLC7A5 overexpression attenuated circZNF124 silencing‐mediated EC malignant progression. Furthermore, SLC7A5 absence inhibited tumor growth in vivo.ConclusionCircZNF124 depletion inhibited EC cell malignancy by miR‐199b‐5p/SLC7A5 pathway, which demonstrated that circZNF124 had the potential as a therapeutic target for EC.

Identification of Vesicle‐Mediated Transport‐Related Genes for Predicting Prognosis, Immunotherapy Response, and Drug Screening in Cervical Cancer

ABSTRACT Background Cervical cancer is one of the most common malignancies among women. Vesicle‐mediated transport mechanisms significantly influence tumor cell behavior through intercellular material exchange. However, prognostic significance in CC patients remains underexplored. Research Design and Methods We identified differentially expressed vesicle‐mediated transport‐related genes from TCGA and GeneCards datasets through differential expression analysis. We constructed a prognostic model using Cox regression and LASSO regression, categorized patients into high‐ and low‐risk groups, and validated the model in the GEO data set. A nomogram integrating clinical features and risk scores demonstrated the model's independent prognostic capability. We analyzed tumor immune cell infiltration, immune checkpoints, and predicted immunotherapy responses in the high‐ and low‐risk groups. Finally, we screened potential drugs for targeting CC and conducted drug‐sensitivity analysis. Results We successfully established a 10‐gene prognostic model based on VMTRGs. The low‐risk group exhibited favorable prognosis, significant immune cell infiltration, and promising immunotherapy response, whereas the high‐risk group showed higher sensitivity to chemotherapeutic agents such as Docetaxel and Paclitaxel. Potential drugs identified for targeting CC patients included Megestrol acetate, Lenvatinib, Adavosertib, and Barasertib. Conclusions The VMTRG‐based prognostic model demonstrates reliable clinical prognostic value and enhances understanding of vesicle‐mediated transport mechanisms in CC.

MicroRNA‐378a‐3p contributes to ovarian cancer progression through downregulating PDIA4

AbstractObjectiveMicroRNAs, as essential players in tumorigenesis, have been demonstrated to have a revolutionary effect on human cancer research. Ovarian cancer is the primary reason of death among gynecologic malignancies. In view of this, it is significant to identify prognostic and predictive markers for treatment of ovarian cancer. The aim of this study was to probe into the effects of miR‐378a‐3p and protein disulfide‐isomerase A4 (PDIA4) on the biological functions of ovarian cancer cells.MethodsmiR‐378a‐3p expression and PDIA4 messenger RNA expression in human ovarian cancer cells, normal human ovarian epithelial cells, and serum of both ovarian cancer patients and healthy people were detected by reverse transcription‐quantitative polymerase chain reaction, and the PDIA4 protein expression was tested by Western blot analysis. Ovarian cancer OVCAR3 and SKOV3 cells were transfected or cotransfected with miR‐378a‐3p mimic or pcDNA3.1‐PDIA4 or their negative control plasmids to explore their roles in biological functions in ovarian cancer cells. Luciferase activity and RIPA assays were implemented to validate the interaction between miR‐378a‐3p and PDIA4. Western blot analysis was utilized to detect phosphatidylinositol‐3 kinase/serine/threonine kinase (PI3K/AKT) signaling pathway‐related protein expression and their phosphate expression levels.ResultsmiR‐378a‐3p was elevated and PDIA4 was decreased in ovarian cancer cells and serum. In addition, miR‐378a‐3p mimic induced ovarian cancer cell growth, while miR‐378a‐3p inhibitor and pcDNA3.1‐PDIA4 presented an inverse trend. pcDNA3.1‐PDIA4 partially eliminated the capabilities of miR‐378a‐3p mimic on ovarian cancer progression. Meanwhile, miR‐378a‐3p was found to negatively regulate PDIA4, and miR‐378a‐3p mimic increased the phosphorylation levels of AKT and PI3K, while pcDNA3.1‐PDIA4 exhibited an opposite tendency. Furthermore, pcDNA3.1‐PDIA4 largely eliminated the functions of miR‐378a‐3p mimic on phosphorylation levels of AKT and PI3K.ConclusionThis study provides evidences that miR‐378a‐3p activates PI3K/AKT signaling pathway by modulating PDIA4 expression, thereby playing a role in promoting the growth of ovarian cancer cells. This study provides novel directions for targeted therapy of ovarian cancer.

Cancer‐testis antigen ACRBP expression and serum immunoreactivity in ovarian cancer: Its association with prognosis

AbstractIntroductionCancer testis (CT) antigens are attractive targets for cancer immunotherapy because of their expression restriction and immunogenicity. The acrosin binding protein (ACRBP) is a member of CT antigens. This study aimed to evaluate ACRBP expression and immunogenicity in ovarian cancer (OC).MethodsThe expression level of ACRBP in OC tissues, normal ovarian tissues, and cell lines was detected via quantitative real‐time polymerase chain reaction (qRT‐PCR) and immunohistochemistry. We determined the levels of ACRBP antigen and antibody in serum samples collected from patients with OC and healthy donors using enzyme‐linked immunosorbent assays (ELISA), the level of ACRBP in cell‐cultured medium was also tested.ResultsACRBP mRNA and protein expressions were upregulated in OC tissues relative to normal tissue, especially highly expressed in epithelial ovarian cancer (EOC). Moreover, ACRBP expression was significantly correlated with International Federation of Gynecology and Obstetrics (FIGO) stage and chemosensitivity. Serological analysis showed that anti‐ACRBP antibody was detected in the sera of 16 of the 56 (28.5%) patients with OC but not in healthy donors. The area under the receiver operating characteristic curve for ACRBP antibody was 0.802 (95% confidence interval [CI]: 0.708–0.876), and the sensitivity and specificity for ACRBP antibody was 85.71% and 55.0%, respectively. Kaplan–Meier analysis revealed that the overall survival (OS) and disease‐free survival (DFS) in OC patients with high ACRBP expression were significantly lower than those with low expression (p = 0.040, p = 0.021). However, ACRBP antibody level was not associated with prognosis.ConclusionACRBP expression was upregulated in OC tissues and induced humoral immune response in patients with OC, suggesting that ACRBP is a potential prognostic biomarker and a target of tumor immunotherapy for OC.

CAR‐NK, a Splendid Strategy for Cancer, Especially for Gynecologic Tumor

ABSTRACTBackgroundNK cells are a class of innate lymphocytes capable of nonspecifically killing tumor cells without MHC restriction or prior sensitization. Recent advancements in biotechnology, particularly the development of chimeric antigen receptors (CAR) and related technologies, have enabled targeted tumor cell elimination. CAR endows NK cells with enhanced functionality, with the extracellular domains typically consisting of single‐chain variable fragments (scFv) for targeting specific antigens. CAR‐NK cells have shown excellent results in several preclinical studies and clinical trials for hematologic malignancies. However, their clinical application in the treatment of solid tumors is still insufficient. Current treatments for gynecological cancers primarily involve surgery, chemotherapy, and radiotherapy, all of which often present substantial side effects and variable efficacy. While CAR‐T cell therapy has shown effectiveness in certain gynecological tumors, its clinical application is hindered by severe side effects, such as Cytokine Release Syndrome (CRS) and Graft‐Versus‐Host Disease (GVHD). CAR‐NK cell therapy offers improved safety profiles in clinical applications.ObjectiveThis review aims to systematically evaluate recent methodological innovations in CAR‐NK engineering and their translational potential in tumor‐targeted treatment, providing valuable insights for clinical trials and studies.MethodsElectronic databases, including PubMed and Web of Science were searched for relevant literature. Keywords are as follows: CAR‐NK cell; Chimeric antigen receptor; Solid tumor; cell therapy; gynecological cancers.ResultsCAR‐NK engineering has innovations such as multi‐targeted CAR design, gene editing for enhanced persistence, and “off‐the‐shelf” CAR‐NK cells compared to CAR‐T cells.ConclusionCAR‐NK cell therapy combines safety and anti‐tumor efficacy, particularly for gynecological cancers.

The relationship of 3′UTR HLA‐G14‐bp insertion/deletion and +3142 C/G polymorphisms and soluble HLA‐G expression with gynecological cancers: An updated meta‐analysis

AbstractObjectivesHuman leukocyte antigen‐G (HLA‐G) is implicated in several cancers and is considered to be an immune checkpoint regulator. We determined the association between polymorphisms in the 3′ untranslated region of HLA‐G and soluble HLA‐G (sHLA‐G) expression with gynecological cancers (GCs).MethodsA meta‐analysis was conducted to examine the association between HLA‐G14‐bp insertion (I)/deletion (D) and +3142C/G polymorphism in GC and to evaluate sHLA‐G expressionResultsWe revealed a significant association between the +3142C/G polymorphism and invasive cervical cancer (ICC) based on the allelic model G versus C (odds ratio [OR] = 0.738, 95% confidence interval [CI] = 0.563–0.966, p = 0.027), dominant GG+GC versus CC (OR = 0.584, 95% CI = 0.395–0.862, p = 0.007), and codominant GG versus CC (OR = 0.527, 95% CI = 0.312–0.891, p = 0.017) models, suggesting that the G allele and GG genotype are protective against ICC. In gynecological precancerous patients with human papillomavirus (HPV) infection, we found that the 14‐bp I/D under the codominant DD versus DI model (OR = 0.492, 95% CI = 0.241–1.004, p = 0.051) was of borderline significance. Soluble HLA‐G levels were significantly higher in patients compared with healthy controls (standardized mean differences [SMD] = 1.434, 95% CI = 0.442–2.526, p = 0.005). Stratification by cancer type revealed that the sHLA‐G levels were significantly increased in cervical cancer (SMD = 4.889, 95% CI = 0.468–9.310, p = 0.030) and in subjects of Asian ethnicity (SMD = 4.889, 95% CI = 0.467–9.309, p = 0.030).ConclusionsHLA‐G14‐bp I/D and +3142 C/G polymorphisms are associated with GC and HPV‐associated cervical cancer. In addition, we found significantly increased sHLA‐G levels in cancer patients. These results provide a basis for further studies in diagnostics and immunotherapy of GC.

Activation of the cGAS–STING signaling pathway in adenomyosis patients

AbstractObjectiveAdenomyosis is characterized by the presence of endometrium or endometrium‐like glands and stroma within the myometrium. In this study, we aimed to investigate whether the cGAS–STING pathway was activated and correlated with clinical outcomes in adenomyosis patients.Materials and MethodsTwenty patients diagnosed with adenomyosis and 10 patients diagnosed with cervical intraepithelial neoplasia grade 3 (CIN‐3) but no adenomyosis were enrolled in this study. Specimens were collected during surgery from August 2017 to December 2017 at Third Xiangya Hospital. The messenger RNA (mRNA) and protein levels of key cGAS–STING pathway factors in uterine tissue were detected by real‐time reverse‐transcription polymerase chain reaction and immunohistochemistry, respectively. The correlations of gene expression and clinical outcomes, including dysmenorrhea and uterine volume, were analyzed.ResultsThe cGAS, STING, TANK‐binding kinase 1 (TBK‐1), interferon‐α (IFN‐α), IFN‐β, and tumor necrosis factor‐α (TNF‐α) mRNA and protein levels in the ectopic endometrial tissue from adenomyosis patients were significantly higher compared with that from the controls in endometrium (p < .05). cGAS and STING gene expression were correlated with TBK‐1, IFN‐β, and TNF‐α expression (p < .05). Importantly, TBK‐1 and TNF‐α expression were correlated with the clinical outcome of dysmenorrhea (p < .05).ConclusionOur study reveals that the cGAS–STING pathway is activated in adenomyosis patients and its activation is subsequently correlated with clinical outcomes, which suggests that the cGAS–STING pathway may contribute to adenomyosis pathogenesis.