Human Mutation

BST2 Drives Epithelial Ovarian Cancer Progression via Macrophage M2 Polarization, Neural Remodeling, and Immunosuppressive Microenvironment Formation

Background Epithelial ovarian cancer (EOC) ranks as the most lethal of gynecological cancers. Despite advances in therapeutic interventions that have marginally extended survival rates, the early detection and management of EOC pose significant hurdles. Consequently, identifying novel therapeutic targets is imperative for enhancing the survival outcomes of patients afflicted with this malignancy. Purpose This research is aimed at exploring the functions of Bone Marrow Stromal Antigen 2 (BST2) in the pathogenesis of EOC and their influence on macrophage polarization, evaluating their viability as targets for immunotherapy. Methods Gene expression profiles and clinical data of EOC patients were retrieved from the TCGA repository to develop prognostic models centered on BST2. The expression patterns of BST2 in HGSOC cell lines were quantified via RT‐qPCR and Western blot analyses. The impact of BST2 on the proliferative, migratory, and invasive capacities of EOC cells was assessed through gene silencing and gene overexpression experiments. Results Elevated levels of BST2 expression were observed in EOC tissues, correlating with adverse prognostic indicators. Enhanced BST2 expression facilitated EOC cell growth, motility, and invasiveness, whereas BST2 suppression mitigated these oncogenic attributes. In vivo assessments revealed that BST2 augmentation modified the macrophage phenotypes within grafted ovarian tumors, with BST2 diminution reversing these effects. Conclusion The findings propose that BST2 acts as a pivotal facilitator in the progression of ovarian carcinoma. The expression metrics of BST2 may serve as prognostic markers for patient outcomes in EOC. These findings suggest that BST2 is a key promoter of ovarian cancer progression, and its expression may serve as a prognostic marker. The mechanisms uncovered, including the modulation of macrophage polarization and neural marker expression, indicate that targeting BST2 represents a potential future strategy for immunotherapy in EOC.

Prediction of Immunotherapy Response and Prognostic Outcomes for Patients With Ovarian Cancer Using PANoptosis‐Related Genes

BackgroundOvarian cancer (OC) is a lethal malignancy often diagnosed at a late stage with frequent recurrence and immunotherapy resistance. PANoptosis is a novel programmed cell death regulating tumors and immunity. We constructed a prognostic model based on PANoptosis‐related genes (PRGs) and evaluated its value for predicting immunotherapy response and survival in OC.MethodsPRGs linked to OC prognosis were identified from public databases, followed by using the STRING database to develop a protein–protein interaction (PPI) network. The LASSO and multivariate Cox regression analyses were used to construct a risk model, and its predictive value was verified by survival analysis, receiver operator characteristic (ROC) curve, and nomogram. Next, we analyzed the immune microenvironment by combining CIBERSORT, MCP‐counter, and ssGSEA algorithms and assessed the response of patients in different risk groups to immunotherapy using TIDE with immune phenotype score (IPS) methods. GSEA was performed to evaluate the activation status of biological pathways between patients in different risk groups. Finally, we verified the expression and potential biological functions of the key genes using quantitative reverse transcription‐PCR (qRT‐PCR), CCK‐8, scratch, and transwell assays.ResultsA PANoptosis‐related risk model for OC was constructed based on eight genes (PIK3CG, CAMK2A, CD38, NFKB1, PSMA4, PSMA8, PSMB1, and STAT4). The model could accurately evaluate the prognostic outcomes for OC patients, showing a high stability across different datasets. High‐risk patients had lower immune cell infiltration, elevated TIDE, and reduced IPS, which suggested weaker immunotherapy responsiveness and therefore a worse prognosis. In addition, pathway analysis showed that the high‐risk group was mainly enriched in tumor progression–related pathways. In vitro, PIK3CG, CAMK2A, NFKB1, PSMA4, and PSMB1 were upregulated in OC cell lines, and knockdown of PIK3CG notably suppressed the proliferative, migratory, and invasive capabilities of OC cells.ConclusionThe PRG model established in this study may contribute to the assessment of immunotherapeutic response and prognosis for OC patients.

Genetic Causal Relationship Between Systemic Lupus Erythematosus and Malignant Tumors of the Female Reproductive System: A GWAS Analysis in European Populations

Background: Systemic lupus erythematosus (SLE) is a multisystem autoimmune disease that primarily affects women of reproductive age. Existing studies have demonstrated complex associations between SLE and various diseases, but its genetic relationship with malignant tumors of the female reproductive system has not been fully elucidated. This study is aimed at exploring the potential genetic associations and shared molecular basis between SLE and female reproductive system malignancies using genome‐wide association studies (GWASs) and cross‐trait analysis.Methods: We selected genetic variants significantly associated with SLE (p < 5 × 10−8) from large‐scale GWAS databases as genetic instruments and applied various statistical methods to analyze the associations between SLE and cervical cancer, endometrial cancer, ovarian cancer, vulvar cancer, vaginal cancer, and uterine cancer. The primary analysis was conducted using inverse variance weighting (IVW), supplemented by Egger regression, weighted median, and weighted mode methods. To control for potential confounders, we performed multivariable analysis while including BMI, estradiol, and CRP as covariates. Additionally, cross‐trait analysis using the association analysis based on subset (ASSET) method was employed to identify shared genetic variants and their effect directions between SLE and uterine cancer.Results: Genetic association analysis showed a significant negative association between SLE and endometrial cancer (OR = 0.972, 95% CI [0.946–0.998], p = 0.038), suggesting that SLE may be associated with a reduced risk of endometrial cancer. For uterine cancer, the weighted median method also indicated a marginally significant negative association (OR = 0.955, 95% CI [0.912–1.000], p = 0.049). Multivariable analysis further confirmed that the protective association between SLE and endometrial cancer remained significant after controlling for BMI, estradiol, and CRP (OR = 0.96, 95% CI [0.93–0.99], p = 0.014). However, no significant association was observed between SLE and cervical cancer, ovarian cancer, vulvar cancer, or vaginal cancer. Cross‐trait analysis identified 193 shared genetic variants between SLE and endometrial cancer and 71 shared variants between SLE and uterine cancer, with rs2442719 and rs3131004 showing consistent effect directions in both comparisons.Conclusion: This study provides genetic epidemiological evidence suggesting that SLE may have a protective effect against endometrial and uterine cancers and identifies potential shared genetic bases. These findings offer new insights into the relationship between SLE and gynecological tumors and may provide references for the prevention and treatment of related diseases.

Single‐Cell and Spatial Transcriptomics Explore Purine Metabolism–Related Prognostic Risk Model and Tumor Immune Microenvironment Modulation in Ovarian Cancer

Background: Ovarian cancer (OC) ranks as the second leading cause of gynecological cancer–related deaths in women globally. Single‐cell and spatial transcriptomics could precisely describe the heterogeneity of OC that affect the clinical treatment.Methods: Single‐cell sequencing and spatial transcriptomics information were from different public datasets. A pseudotime analysis of cellular developmental pathways, score single‐cell gene sets, and cell activity ratings in each metabolic pathway were performed. A prognostic model was created using univariate regression analysis, LASSO, and multivariate regression analysis. Finally, the immune microenvironment and immunotherapeutic effects were analyzed for their association with purine metabolism activity. Finally, RT‐qPCR was used to estimate the mRNA level of OC in cell lines.Results: We observed a higher purine metabolism score by a signature of 12 purine metabolism–related gene in tumor cells. When compared with fibroblasts, epithelial cells with high scores displayed more intense TGF‐β signaling pathway activity. Forty‐four differentially expressed purine metabolism–related genes were identified to be substantially expressed in the tumor’s core region and were closely linked to purine and pyrimidine metabolic activities. Low‐risk population had higher immune infiltration level and immunotherapy results. The NME6+ epithelial cell high‐expression group had a greater prognosis and showed a negative connection with the tumor immune dysfunction and exclusion score and cancer‐associated fibroblast cell concentration.Conclusion: Purine metabolism was a predictor for OC patients’ prognosis. The presence of positive NME6 expression in epithelial cells emerges as a protective factor for OC patients, presenting a possible therapeutic target for personalized treatment.

Prognostic Value of Ubiquitination‐Related Genes in Ovarian Cancer and Their Correlation With Tumor Immunity

Numerous studies have emphasized the importance of the ubiquitin–proteasome system (UPS) in the malignant progression of ovarian cancer (OC). However, whether ubiquitination‐related genes (UbRGs) can be used to predict the prognosis of OC remains to be revealed. Patients with OC were divided into two clusters based on the expression of UbRGs, and prognosis was compared between the two clusters. A prognostic model was established based on UbRGs, and its predictive efficiency was validated using Kaplan–Meier (K–M) curves, receiver operating characteristic (ROC) curves, and a nomogram. Immune infiltration and gene mutation analyses were used to examine the effects of UbRGs on the prognosis of OC. The prognostic model served as a valid and independent predictor of OC prognosis. Immune infiltration revealed that the unique immune microenvironment of OC was regulated by UbRGs. Gene mutation analysis indicates that UbRGs likely influence OC malignant behavior by modulating gene mutation patterns. In addition, Ube2j1 was found to play an important role in regulating the malignant progression of OC. Furthermore, the mechanism by which Ube2j1 modulates the OC phenotype and reshapes its immune microenvironment via the JAK2/STAT3/PD‐L1 pathway was elucidated, providing novel insights into the potential for ubiquitination‐based immunotherapy in OC. This study provides novel insights into precision immunotherapy based on UbRGs in OC. The UbRGs‐based prognostic model may help to provide novel insights for the application of ubiquitination‐based immunotherapy in OC.

Causal Relationships Between Pregnancy, Menstrual History, and Endometrial Cancer With Mediating Effects of Metabolism‐Related Traits

Background Periods and pregnancy may affect the development of endometrial cancer by affecting the secretion of sex hormones, but the causal relationship is not clear, and its mediating factors need to be explored. Methods In this study, multivariable Mendelian randomization was used to analyze summary statistics of genome‐wide association studies of European ancestry, to evaluate the effect of 10 period‐ or pregnancy‐related factors on endometrial cancer. In addition, we performed the heterogeneity test and pleiotropy test to analyze the sensitivity. Because of the effect of sex hormones on body metabolism and the relationship between metabolism‐related traits and cancer, we explored the mediating effect of metabolism‐related traits by two‐step Mendelian randomization. Results This study showed that age at menarche ( p = 1.21e − 05; OR = 0.6852; 95% CI: 0.5784–0.8116), age at menopause ( p = 0.00098; OR = 1.242; 95% CI: 1.0919–1.4127), and sex hormone–binding globulin (SHBG) levels ( p = 7.4e − 07; OR = 0.5914; 95% CI: 0.4804–0.7281) have an independent causal relationship with the incidence of endometrial cancer. Moreover, several obesity‐related traits play a mediating role in the causal relationship between age at menarche and endometrial cancer. The mediators and their mediating effects are BMI (55.54%), obesity (30.37%), waist circumference preference (27.67%), body fat percentage (17.61%), and waist‐to‐hip ratio (14.82%). These results are robust to sensitivity analysis. Conclusion This study demonstrated the independent effect of pregnancy‐ and period‐related factors on endometrial cancer and suggested that avoiding obesity may be an effective method to prevent endometrial cancer for patients with premature menarche.

FCGR2B + Macrophages as a Critical Node Linking Ferroptosis and Immunosuppression: A Multiomics Framework for Prognosis and Therapy in High‐Grade Serous Ovarian Cancer

Background High‐grade serous ovarian cancer (HGSOC) is characterized by a complex tumor microenvironment and poor prognosis, yet the roles of specific tumor‐associated macrophages (TAMs) subpopulations in driving disease progression remain elusive. Methods This study evaluated the prognostic relevance of FCGR2B in HGSOC. Single‐cell RNA sequencing identified FCGR2B + TAMs as a distinct macrophage subpopulation with unique transcriptional features. Integrative analyses combining single‐cell and bulk differentially expressed genes, macrophage‐associated modules, and ferroptosis‐related gene sets identified 26 candidate prognostic genes, from which a four‐gene signature ( CRYAB , PLAUR , EREG , and C5AR1 ) was derived to construct the prognostic risk model. The model was validated in an independent cohort. Immune infiltration, single‐cell trajectory, copy number variation, and drug–gene associations were analyzed to explore the molecular and therapeutic implications of risk stratification. Results HGSOC patients classified as high risk exhibited poorer survival outcomes, increased infiltration of M2‐like macrophages, elevated expression of immune checkpoints, and enrichment of immune‐ and ferroptosis‐related pathways. Trajectory and copy number variation analyses revealed stage‐specific gene expression patterns and amplification‐associated regulation. Drug–gene association analyses further suggested that high‐risk patients may be more responsive to targeted therapies and proteasome inhibitors, whereas low‐risk patients may benefit from conventional chemotherapy. Conclusion FCGR2B + TAMs are closely linked to HGSOC progression, and the proposed prognostic model based on FCGR2B + TAMs provides predictive value and potential therapeutic insights for patient stratification.

Case–case analysis addressing ascertainment bias for multigene panel testing implicates BRCA1 and PALB2 in endometrial cancer

Hereditary endometrial cancer (EC) is most commonly attributed to pathogenic variants in mismatch repair genes. Evidence supports the existence of additional genetic risk factors in the context of multiple cancer diagnoses and/or family history of EC. EC patients (n = 5292) referred for diagnostic multigene cancer panel testing were annotated for presence of a pathogenic gene variant; personal history of prior, concurrent, or subsequent cancer of another type; reported family history of Lynch syndrome or EC. The Pearson χ

A Novel Alu Element Insertion in ATM Induces Exon Skipping in Suspected HBOC Patients

The vast majority of patients at risk of hereditary breast and/or ovarian cancer (HBOC) syndrome remain without a molecular diagnosis after routine genetic testing. One type of genomic alteration that is commonly missed by diagnostic pipelines is mobile element insertions (MEIs). Here, we reanalyzed multigene panel data from suspected HBOC patients using the MEI detection tool Mobster. A novel Alu element insertion in ATM intron 54 (ATM:c.8010+30_8010+31insAluYa5) was identified as a potential contributing factor in seven patients. Transcript analysis of patient-derived RNA from three heterozygous carriers revealed exon 54 skipping in 38% of total ATM transcripts. To manifest the direct association between the Alu element insertion and the aberrant splice pattern, HEK293T and MCF7 cells were transfected with wild-type or Alu element-carrying minigene constructs. On average, 77% of plasmid-derived transcripts lacked exon 54 in the presence of the Alu element insertion compared to only 4.7% of transcripts expressed by the wild-type minigene. These results strongly suggest ATM:c.8010+30_8010+31insAluYa5 as the main driver of ATM exon 54 skipping. Since this exon loss is predicted to cause a frameshift and a premature stop codon, mutant transcripts are unlikely to translate into functional proteins. Based on its estimated frequency of up to 0.05% in control populations, we propose to consider ATM:c.8010+30_8010+31insAluYa5 in suspected HBOC patients and to clarify its role in carcinogenesis through future epidemiological and functional analyses. Generally, the implementation of MEI detection tools in diagnostic sequencing pipelines could increase the diagnostic yield, as MEIs are likely underestimated contributors to genetic diseases.

Constitutional BRCA1 Epimutations: A Key for Understanding Basal‐Like Breast and High‐Grade Serous Ovarian Cancer

Germline pathogenic genetic variants in the BRCA1 and BRCA2 genes are the most frequent causes of familial breast and ovarian cancer. Contrasting BRCA2, epimutations in the BRCA1 gene are frequently detected in tissue from triple‐negative breast (TNBC) and high‐grade serous ovarian cancers (HGSOC). While studies over the last decade have reported BRCA1 epimutations in white blood cells (WBC) from breast and ovarian cancer patients, the potential hazard ratio for incident TNBC and HGSOC was not formally assessed until recently.Conducting a prospective nested case‐control study on women participating in the American Women’s Health Initiative Study, we provided firm evidence that mosaic WBC BRCA1 epimutations, even at allele frequencies < 0.1%, are associated with a significantly increased risk of both incident HGSOC and TNBC > 5 years after WBC collection. In a second study assessing BRCA1 epimutations in WBC and matched tumor samples from TNBC, our results indicated such epimutations to be the underlying cause of around 20% of TNBC, far exceeding the percentage of cases carrying BRCA1 germline pathogenic genetic variants.We detected primary constitutional BRCA1 epimutations in tissues derived from all three germ layers. They occur independently of BRCA1 promoter haplotypes but are present on the same allele in all WBC within affected individuals. Moreover, epimutations are consistently found on the same allele in normal and tumor breast tissue as well as in WBC. This finding, together with BRCA1 epimutations detected in WBC from newborns, strongly indicates an early embryonic event with clonal expansion affecting all germ layers.Future work in the field must lead to an understanding of exactly when and how the BRCA1 epimutations occur and, most importantly, whether primary constitutional epimutations in genes other than BRCA1 may cause an elevated risk of other cancer types.

A decade of RAD51C and RAD51D germline variants in cancer

Defects in DNA repair genes have been extensively associated with cancer susceptibility. Germline pathogenic variants (GPV) in genes involved in homologous recombination repair pathways predispose to cancers arising mainly in the breast and ovary, but also other tissues. The RAD51 paralogs RAD51C and RAD51D were included in this group 10 years ago when germline variants were associated with non-BRCA1/2 familial ovarian cancer. Here, we have reviewed the landscape of RAD51C and RAD51D germline variants in cancer reported in the literature during the last decade, integrating this list with variants identified by in-house patient screening. A comprehensive catalog of 341 variants that have been classified applying ACMG/AMP criteria has been generated pinpointing the existence of recurrent variants in both genes. Recurrent variants have been extensively discussed compiling data on population frequencies and functional characterization if available, highlighting variants that have not been fully characterized yet to properly establish their pathogenicity. Finally, we have complemented this data with relevant information regarding the conservation of mutated residues among RAD51 paralogs and modeling of putative hotspot areas, which contributes to generating an exhaustive update on these two cancer predisposition genes.

Oncological Genetic Counseling in Hereditary Breast and Ovarian Cancers and Lynch Syndrome High‐Risk Subjects: Evaluation of Efficacy and Outcomes Using the Genomics Outcome Scale

Background Validated tools assessing oncological genetic counseling (OGC) quality are lacking. Methods We assessed OCG effectiveness using italian‐translated version of the Genomics Outcome Scale (GOS) questionnaire. Clinical variables were collected and their association with different answers was assessed by Fisher′s exact test or Chi‐square test for either dichotomous or other categorical variables, respectively, with level of statistical significance p = 0.05. Results Between November 2024 and February 2025, 209 subjects who received the complete OGC program at Our Center responded to the questionnaire; median age was 56 years (25–81). Most (76%) had breast cancer, 72% received a negative test, 15% positive test, and 13% noninformative test with variant of unknown significance (VUS). Most patients answered affirmatively to Question 1, focused on OGC understanding: age ( p = 0.0181) and education ( p = 0.0028) yielded different answers. Question 2, assessing relatives risk understanding, was answered completely/partially affirmative by 94% of subjects: test result (negative noninformative vs. positive vs. VUS) was associated ( p = 0.0175) with different answers. To Question 3, related to concern, 65% confirmed their worry: education ( p = 0.0392) and cancer type ( p = 0.0128) yielded different answers. In Question 4, focused on surveillance understanding, 77% declared full or partial awareness, regardless of examined factors. In Question 5, enquiring decisional ability for themselves or family members, 72% stated they were completely/partially able to make decisions. Education ( p = 0.0287) and genetic test result ( p = 0.0090) yielded different answers. In Question 6, reflecting future planning, 69% responded completely/almost completely affirmatively, 17% were uncertain, and 14% responded partially/completely negatively, regardless of examined clinical factors. Conclusions: GOS questionnaire confirms that OGC is useful and effective to inform patients about their condition, surveillance, and prevention. Higher levels of empowerment were seen in younger patients and those with higher education.

IPO9 Promotes Ovarian Cancer Progression by Suppressing HMOX1‐Dependent Ferroptosis

Ovarian cancer (OC) poses a significant threat to women’s health, with current treatment strategies remaining suboptimal, necessitating the exploration of novel therapeutic targets and immune microenvironment dynamics. This study integrates multiomics data from TCGA, GEO, and IEU‐Open‐GWAS, employing scRNA‐seq, scPagwas, BayesPrism, and WGCNA to identify key cell subpopulations and genes, followed by functional validation through EdU, colony formation, Transwell assays, and ferroptosis markers (MDA, ROS, and ferrous ions). Results reveal MALAT1 + epithelial cells as a core cell subpopulation in OC, with higher abundance correlating with shorter overall survival, suppressed immune microenvironments, and potential immunotherapy resistance, while their infiltration levels are closely associated with OC immune dynamics and somatic mutations. Further analysis identifies IPO9 as a core gene upregulated in OC, promoting tumor progression by inhibiting HMOX1‐dependent ferroptosis. These findings highlight MALAT1 + epithelial cells as drivers of immune suppression in OC and propose IPO9 as a promising therapeutic target, offering new avenues for immunotherapy development.

Characterization of Copy Number Variants in Hereditary Cancer Patients Through NGS Shows a Distinctive PALB2 Contribution to the Diagnostic Yield

The extensive use of next‐generation sequencing (NGS) multi‐gene panels and advanced analysis algorithms have led to the identification of numerous genetic variants associated with breast, ovarian, and pancreatic cancer. Copynumber variations (CNVs), defined as deletions and duplications of specific DNA regions, account for up to 10% of pathogenic variants and can affect any of the cancer‐predisposing genes. Despite this, CNVs’ contribution beyond BRCA1 and BRCA2 remains underexplored. This observational study analyzed data from 2949 patients, primarily affected by breast or ovarian cancer, who underwent NGS testing with a 22‐gene hereditary cancer panel between 2018 and 2023, with a focus on CNV results. In line with comparison studies, a total diagnostic yield of 14.8% was observed with pathogenic variants in BRCA1, BRCA2, CHEK2, ATM, and PALB2 accounting for most of positive findings. In contrast, CNVs were found in 1.4% of patients, displaying a peculiar distribution pattern. PALB2 exhibited the highest frequency of pathogenic CNVs (66.7%), representing 62.2% of all PALB2 pathogenic variants. Notably, 24 out of 28 PALB2 CNV carriers shared the deletion of Exon 11. Further investigations revealed identical breakpoints and common geographical origins, and moreover, the same haplotype for some of the families suggests a relatively recent founder effect. Simultaneous sequence and copy number analyses resulted in likely higher positive predictive value of the test and, more interestingly, disclosed an unforeseen single contribution of CNVs in PALB2 gene, confirming geography as a key factor in shaping human genetic variations.