Gudaitienė, Jurgita

Background and Objectives Germline pathogenic or likely pathogenic (P/LP) variants account for 5–10% of all breast cancer (BC) cases, with a higher prevalence of 10–20% observed in specific BC molecular subtypes, such as triple-negative breast cancer (TNBC). P/LP variants in established BC susceptibility genes are associated with an increased risk of BC and may contribute to distinct tumor phenotypes and clinical outcomes. This study aimed to evaluate the distribution of germline P/LP variants and their associations with clinicopathological features and clinical outcomes in BC patients. Material and Method A total of 85 female patients with stage I-II BC, including hormone receptor-positive (HR+)/HER2-negative (HER2−) (n=60) and TNBC (n=25), were included. Inclusion criteria required complete clinicopathological data (age at diagnosis, tumor size, tumor histological grade, lymph node involvement, and clinical outcomes) and absence of clinically significant comorbidities. Exome sequencing of germline DNA from peripheral blood was performed to identify variants in 13 BC susceptibility genes (BRCA1, BRCA2, CDH1, PTEN, STK11, TP53, ATM, CHEK2, PALB2, RAD51C, RAD51D, BARD1, and NF1). Variant analysis was conducted using the Franklin platform (Qiagen), and pathogenicity was re-evaluated according to current ENIGMA, ClinGen, and CanVIG guidelines. For statistical analysis, patients were stratified into P/LP variant carriers and non-carriers. Associations with clinicopathological features and clinical outcomes were assessed, with age included as a continuous covariate for adjustment. The study was approved by the Kaunas Regional Biomedical Research Ethical Committee (Nos. BE-2-10 and P1-BE-2-10/2014). Results Overall, 11 distinct P/LP variants were identified (six in BRCA1, three in CHEK2, and one each in BRCA2 and ATM). These variants were detected in 20/85 patients (23.5%), including 10/60 (16.7%) with HR+/HER2− BC and 10/25 (40%) with TNBC. Only one HR+/HER2− BC patient carried two variants; all others had a single variant. P/LP variants were significantly associated with BC molecular subtype (p=0.021), with carriers more likely to have TNBC (OR=3.33, 95% CI 1.17−9.52, p=0.025), although this was not significant after age adjustment (p=0.158). Carrier status was also associated with tumor grade (p=0.039), disease progression (p=0.007), and distant metastasis (p=0.035), but only the association for increased risk of disease progression for patients carrying P/LP variants remained significant after age adjustment (OR=3.09, 95% CI 1.01–9.43, p=0.048). In the TNBC subgroup, P/LP variant carrier status was statistically associated with lymph node involvement (p=0.009), with carriers having higher odds of lymph node involvement, even after age adjustment (OR=15.18, 95% CI 2.01–113.35, p=0.008), although this finding should be interpreted cautiously due to the small sample size. Survival analysis showed a significant association between P/LP variant carrier status and progression-free survival (PFS) in the overall cohort (p=0.011), which was likely driven by the HR+/HER2− BC subgroup (p=0.030) but not TNBC (p=0.154). In HR+/HER2− patients, carriers had a higher risk of shorter PFS in univariate analysis (HR=2.8, 95% CI 1.05–7.54, p=0.039), which was not significant after age adjustment (p=0.084). For metastasis-free survival (MFS), a significant difference was observed in the overall cohort (p = 0.039), but not in subtype-specific analyses. Although carriers showed a higher risk of shorter MFS in univariate analysis (HR=2.41, 95% CI 1.02– 5.72, p=0.046), this did not remain significant after adjustment for age (p=0.073). Conclusions and Recommendations Germline P/LP variants were identified in a substantial proportion of BC patients, with a higher prevalence in TNBC. Although carrier status was associated with more aggressive disease features, particularly disease progression, most associations did not remain significant after adjustment for age. These findings suggest that P/LP variants may contribute to BC heterogeneity, although their independent prognostic value appears limited and requires further investigation in larger cohorts.

Background and Objectives Breast cancer (BC) is a biologically heterogeneous disease with diverse molecular subtypes and clinical outcomes, strongly influenced by genetic factors. ATP-binding cassette (ABC) transporters play an important role in transmembrane processes, including drug efflux, cellular detoxification, and the maintenance of intracellular homeostasis. Variants in genes such as ABCB1 and ABCB8 may contribute to interindividual differences in tumor behavior, influencing clinicopathological characteristics and potentially affecting disease progression and treatment response. This study evaluated the association of two single nucleotide polymorphisms (SNPs), ABCB1 rs1128503 and ABCB8 rs56198402, with BC clinicopathological features. The findings aim to clarify their role in tumor biology and assess their potential as prognostic biomarkers, contributing to a better understanding of the genetic background of BC heterogeneity. Material and Method The study population comprised 170 female patients with histopathologically confirmed early-stage primary BC, aged 30 to 74 years (median age: 46 years). Clinical and tumor pathomorphological data, including tumor size, lymph node involvement, presence of metastases, degree of differentiation, estrogen (ER) and progesterone (PR) receptor status, human epidermal growth factor receptor 2 (HER2) status, disease progression, and mortality, were obtained from oncologists and the follow-up period extended until November 30, 2024. Genomic DNA was extracted from peripheral blood leukocytes using a column-based commercial DNA isolation kit in accordance with the manufacturer’s instructions. Genotyping of ABCB1 rs1128503 and ABCB8 rs56198402 was performed using TaqMan probe-based assays on the QuantStudio™ 3 Real-Time PCR System. The strength of associations between SNPs and BC clinicopathological characteristics was estimated using crude odds ratios (ORs) and corresponding 95% confidence intervals (CIs). Survival outcomes were assessed using Kaplan-Meier analysis. Statistical analysis was conducted using IBM SPSS Statistics, version 30.0.0.0, and a p-value < 0.05 was considered statistically significant. The study protocol was approved by the Kaunas Regional Biomedical Research Ethics Committee (approval numbers BE-2-10 and P1-BE-2-10/2014). Results Significant associations were identified between ABCB1 rs1128503 and PR status, metastatic disease, disease progression, and mortality. Specifically, GA genotype carriers had higher odds of progesterone receptor positivity than GG (OR=2.038; 95% CI 1.016–4.091; p=0.045), which was corroborated by allele-based analysis, showing similarly increased odds among A allele carriers (OR=1.986; 95% CI 1.034–3.816; p=0.039). Conversely, GA carriers demonstrated reduced odds of metastasis (OR=0.301; 95% CI 0.122–0.742; p=0.009), disease progression (OR=0.387; 95% CI 0.171–0.877; p=0.023), and mortality (OR=0.361; 95% CI 0.144–0.907; p=0.030). Furthermore, tumor size and lymph node involvement were also significantly associated (p-value < 0.05) with the studied genotype, indicating a protective effect against more advanced disease features. A significant difference in progression-free survival was observed across ABCB1 rs1128503 genotypes, with the GA genotype associated with more favorable outcomes (p-value < 0.05). No significant associations were found between the ABCB8 rs56198402 and clinicopathological characteristics of BC. Conclusions and Recommendations The ABCB1 rs1128503 was significantly associated with several breast cancer clinicopathological features, including progesterone receptor status, metastasis, disease progression, and progression-free survival, suggesting a potential protective effect of the GA genotype. In contrast, no significant associations were found for the ABCB8 rs56198402 variant. Further studies with larger cohorts are needed to confirm these findings and clarify the biological role of ABCB1 variants in breast cancer progression and prognosis.

Background and Objectives Breast cancer is a leading cause of cancer-related deaths globally. According to WHO data, in 2022, BC accounted for 6.8% of cancer-related deaths worldwide and 6.2% of such deaths within the Lithuanian population. One of the mechanisms underlying BC is the influence of genetic variants. High-frequency, low-penetrance genetic variants, including single nucleotide polymorphisms (SNPs), are more frequently linked to sporadic breast cancer. The epithelial-to-mesenchymal transition (EMT) is a cellular process that plays a role in embryonic development, wound healing, and tumor formation. EMT is crucial in invasion and subsequent metastasis. Snail1, encoded by the SNAI1 gene, is one of the key factors triggering EMT. In this study, we focused on SNAI1 rs8120550, rs1543442, and rs73113648 polymorphisms and their impact on various pathomorphological features and patients’ prognosis within the Lithuanian breast cancer cohort. Material and Method A total of 199 breast cancer patients were studied. The inclusion criteria were as follows: confirmed diagnosis, early BC stage (I–II), and complete medical documentation. Clinicopathological information was collected from medical records in collaboration with oncologists. Polymorphisms of interest were analyzed with real-time PCR-based genotyping. Statistical analysis was performed using SPSS (31.0.0.0). A pvalue < 0.05 was considered statistically significant. The study received approval from the Kaunas Regional Biomedical Research Ethical Committee (no. BE-2-10, no. P1- BE-2-10/2014,), and written informed consent was obtained from all participants. Results Pearson’s chi‑square analysis revealed significant associations between SNAI1 rs8120550 and disease progression (p = 0.011) and metastasis (p = 0.008). Additionally, rs73113648 was significantly associated with age group (p = 0.041), disease progression (p = 0.002), and metastasis (p = 0.002). In univariate logistic regression analysis, for SNAI1 rs73113648, the TT genotype was significantly associated with older age at diagnosis, showing lower odds compared with the CC genotype (OR = 0.247, p = 0.036). In the analysis of clinical outcomes for rs73113648, the TT genotype was statistically associated with higher odds of progression and metastasis compared with CC genotype (OR = 6.207, p = 0.001; OR = 5.952, p = 0.002, respectively). Rs8120550 demonstrated significant associations with disease progression and metastasis, where the AG genotype was associated with significantly higher odds than the AA genotype (OR = 2.718, p = 0.005; OR = 2.872, p = 0.005, respectively). The following are results after Kaplan-Meier method, which was used to evaluate the association between SNPs and PFS, MFS and OS: rs8120550 showed significant results for PFS (p = 0.012) and for MFS (p = 0.009), while rs73113648 demonstrated statistically significant associations with PFS (p = 0.001) and MFS (p = 0.004). Rs1543442 demonstrated no significant associations with any survival outcomes. In univariate Cox regression analysis, rs8120550 (AG versus AA) were associated with shorter PFS (HR = 2.323, p = 0.006) and MFS (HR = 2.529, p = 0.006), while rs73113648 (TT versus CC) was linked to even shorter PFS (HR = 3.650, p < 0.001) and MFS (HR = 3.567, p = 0.002). Conclusions and Recommendations The study revealed that SNAI1 rs8120550 and rs73113648 were significantly associated with an increased risk of disease progression and metastasis. Both SNPs were also associated with poorer PFS and MFS, with rs73113648 showing the strongest effect. These findings suggest that SNAI1 polymorphisms may have prognostic value for clinical outcomes in early-stage BC, but further studies involving a larger, independent cohort, are recommended to confirm our findings.

Background and Objectives Breast cancer (BC) is a heterogenous disease characterized by substantial variability in tumor biology and clinical behavior. This heterogeneity is reflected in clinicopathological features, including hormone receptor status, human epidermal growth factor receptor 2 (HER2) expression, tumor histological grade, tumor size, and disease stage, which describe tumor features and are key determinants of prognosis. Single nucleotide polymorphisms (SNPs) have been proposed as potential contributors to inter-individual differences in these tumor features and clinical outcomes; however, their role in early-stage BC remains insufficiently defined. In this study, polymorphisms in the RRP1B gene, specifically rs2838342 and rs2051407, were investigated for their potential associations with BC features and outcomes. Material and Method Study population. A total of 191 adult female patients with primary stage I-II BC were enrolled. All participants provided written informed consent, and the study was approved by the Kaunas Regional Ethics Committee for Biomedical Research (nos. BE-2-10 and P1-BE-2-10/2014). Peripheral blood samples were collected for genetic analysis. DNA extraction and genotyping. Genomic DNA was extracted from peripheral blood using a spin column-based extraction method with a commercially available kit. SNP genotyping was performed using TaqMan allelic discrimination assays on a Quanstudio 3 Real-Time PCR System. Statistical analysis. Associations between SNPs and clinicopathological features as well as binary clinical outcomes (disease progression, metastasis status, and mortality status) were evaluated using logistic regression under an additive genetic model. Both univariate and age-adjusted multivariable analyses were performed. Sensitivity analyses using alternative genetic models were conducted for selected SNPoutcome associations showing non-linear patterns in genotype distributions. Progression-free, overall, and metastasis-free survival (MFS) were analyzed using Kaplan-Meier with log-rank tests and Cox proportional hazards regression models under an additive genetic model. Multivariable Cox models were adjusted for established clinicopathological prognostic factors. To account for multiple testing, the Benjamini-Hochberg false discovery rate (FDR) correction was applied separately for clinicopathological features (n = 12), binary clinical outcomes (n = 6), and Cox models (n = 6), using p-values from fully adjusted additive models. Kaplan-Meier analyses were considered descriptive and were not included in correction. Adjusted p-values (q-values) < 0.05 were considered significant. All statistical analyses were performed using IBM SPSS Statistics 30.0.0.0. Results Genotype distributions were as follows: for rs2838342, AA 30.9% (n = 59), AG 51.8%, (n = 99), and GG 17.3% (n = 33); for rs2051407, CC 36.6% (n = 70), CT 45.5 % (n = 87), and TT 17.8% (n = 34). The minor allele frequencies were 43.2% for rs2838342 and 40.6% for rs2051407. Genotype frequencies for both SNPs were consistent with Hardy-Weinberg equilibrium (p > 0.05). In age-adjusted additive logistic regression models, rs2838342 was associated with tumor size (p = 0.022) and histological grade (p = 0.032), although neither association remained significant after correction for multiple testing. Under a recessive genetic model, rs2838342 was associated with estrogen receptor (ER) status after adjustment for age group (OR = 0.38, 95% CI 0.17-0.85, p = 0.018), suggesting a lower likelihood of ER positivity among GG homozygotes. A similar nonsignificant trend under the same genetic model was observed for rs2051407 (p = 0.072). Kaplan-Meier analyses showed no statistically significant differences in survival outcomes across genotype groups. Consistently, no associations were observed in Cox regression analyses under unadjusted or multivariable-adjusted models accounting for age and clinicopathological variables. All results remained non-significant after Benjamini-Hochberg FDR correction. Conclusions and Recommendations No statistically significant associations were observed between the studied SNPs and clinicopathological or survival outcomes after multiple testing correction. However, nominal and model-specific associations for rs2838342 indicate a potential role in tumor biology requiring validation in larger, independent cohorts.

Background: Pathogenic and likely pathogenic germline variants in genes associated with breast cancer (BC) play an important role in the development, progression, and response to therapy. Therefore, the identification of these variants can provide valuable insights into the mechanisms of BC pathogenesis and improve early disease detection, treatment strategies, and patient outcomes. Material and Methods: Whole-exome sequencing (WES) was performed on 52 women diagnosed with luminal A or triple-negative BC at the Hospital of Lithuanian University of Health Sciences Kaunas Clinics. The study used genomic DNA extracted from peripheral blood samples. WES data were analyzed using the Franklin platform by Genoox. Pathogenic and likely pathogenic variants were assessed in 81 genes, as defined by the ACMG Secondary Findings V3.2 (2023), and in 18 genes associated with BC. Results: A total of ten pathogenic and four likely pathogenic germline variants were identified in the study group. Of these, five variants were detected in the BRCA1 gene, three in the BRCA2 and CHECK2 genes, and one each in the NBN, ATM, and MUTYH genes. Overall, all variants were found in 19 patients, each of whom had only one clinically significant variant. Nine different variants were identified in patients with luminal A breast cancer, and four - in triple-negative breast cancer. Only one pathogenetic variant was identified in both groups. Conclusion: In conclusion, 14 clinically significant germline variants were identified in patients with luminal A or triple-negative breast cancer that may potentially affect tumor pathogenesis.

Background and Objectives Breast cancer (BC) remains the most prevalent malignancy among women worldwide. Despite advances in early diagnosis and treatment, tumor heterogeneity and its complex pathogenesis continue to present significant challenges, contributing to high cancerrelated mortality. While germline variants are known to influence BC susceptibility, recent studies suggest that variants in genes regulating key cellular processes, such as the cell cycle, may also affect tumor phenotype, disease progression, and response to therapy. However, the role of these variants remains incompletely understood. Thus, the aim of this study was to analyze germline variants in cell cycle-related genes using whole-exome sequencing (WES) and evaluate their prognostic value in a group of BC patients. Material and Method This study included 52 patients diagnosed with either luminal A or triple-negative breast cancer from the Hospital of Lithuanian University of Health Sciences Kaunas Clinics. All participants were selected based on predefined criteria, including confirmed histopathological diagnosis, comprehensive medical data (age at diagnosis, tumor size (T), grade (G), lymph node involvement (N), estrogen and progesterone receptor status, presence of metastases, disease progression, mortality, and clinical outcomes), and absence of significant comorbidities. Peripheral blood samples were collected for genomic DNA extraction, followed by WES. Germline variant analysis was performed using the Franklin by Genoox platform. This study focused on missense variants with a minor allele frequency (MAF) of 5–20% (according to the 1000 Genomes Project) in 55 genes involved in cell cycle regulation (e.g., CDKN1A, ATM, AURKA, and others). Statistical analysis, including association and survival analysis, was performed using IBM SPSS Statistics version 30.0.0.0. A P-value of less than 0.05 was considered statistically significant. The study was approved by the Kaunas Regional Biomedical Research Ethics Committee (Approval Nos. BE-2-10 and P1-BE2-10/2014). Results In this study, germline variants of ATM rs1801516, ATR rs2229032, and TGFB1 rs1800471 were identified using WES analysis. The results showed that the frequency of genotypes was as follows: GG – 61.5%, GA – 36.5%, and AA – 1.9% for ATM rs1801516; CC – 75.0%, CT – 21.2%, and TT – 3.8% for ATR rs2229032; and CC – 96.2%, CG – 3.8%, and GG – 0.0% for TGFB1 rs1800471. All variants were distributed according to the Hardy-Weinberg equilibrium (p>0.05). The association analysis showed that ATM rs1801516 was associated with death (p<0.047); however, logistic regression analysis showed that the association was nonsignificant in both univariate and multivariate models. Survival analysis revealed that the same variant was associated with PFS, MFS, and OS (p<0.001). It was established that patients presenting the AA genotype were more likely to have a longer PFS, MFS, and OS (HR = 32.038, HR = 29.586, HR=60.698, respectively; p<0.05). The results remained statistically significant in multivariate analysis. Neither the ATR rs2229032 nor TGFB1 rs1800471 demonstrated any significant results in association and survival analysis. Conclusions and Recommendations In conclusion, the ATM rs1801516 was found to be significant for mortality and clinical outcomes in breast cancer patients and may have potential as a prognostic biomarker. However, due to the limited sample size, further studies with larger cohort are required to confirm these findings.

Background and objectives The ABCB8, a mitochondrial ATP-binding cassette transporter, is critical in regulating iron homeostasis and protecting cells from oxidative stress. Polymorphisms in the ABCB8 gene, including rs17545756 and rs4148844, are likely to affect gene expression and protein functionality, thereby impacting cancer risk. In breast cancer (BC), these variants can affect tumor behavior by modifying mitochondrial function, metabolic pathways, or therapeutic resistance. This study aims to analyze the association of ABCB8 polymorphisms rs17545756 and rs4148844 with BC by evaluating their correlation with tumor phenotype and disease prognosis in BC patients. Understanding these associations may provide insights into the molecular mechanisms of BC progression and support the development of more targeted diagnostic and therapeutic strategies. Material and Method In this study, we analyzed ABCB8 rs4148844 and rs17545756 polymorphisms in 170 patients with BC. Medical information such as age at diagnosis, tumor size, grade, lymph node status, estrogen (ER), progesterone (PR), human epidermal growth factor 2 (HER2) receptor status, metastatic status, disease progression, and death was collected. Genomic DNA was isolated from peripheral blood leukocytes using a commercially available DNA extraction kit (Thermo Fisher Scientific Baltics, Vilnius, Lithuania). The polymorphisms mentioned were assessed using TaqMan (Applied Biosystems Europe BV, UK Branch, Warrington, Cheshire, UK) according to the manufacturer’s instructions on the QuantStudio 3 real-time PCR system (Applied Biosystems, Foster City, CA, USA). The statistical analysis was done with Statistical Package for the Social Sciences (SPSS) 25.0 for Windows. The study was approved by the Kaunas Regional Biomedical Research Ethical Committee (protocol No. 2024- BEC2-137). Results The association between ABCB8 rs17545756 and lymph node involvement was determined (χ2 (2) = 5.479, p = 0.040). Logistic regression analysis revealed that individuals carrying the CT genotype had a significantly higher risk of lymph node metastasis compared to those with the CC genotype (OR = 2.909, 95% CI 1.003–8.439, p = 0.049). Moreover, T allele carriers (vs. non-carriers) had a 3.200-fold increased risk of lymph node metastasis (OR = 3.200, 95% CI 1.121-9.131, p = 0.030). The association between ABCB8 rs17545765 and lymph node metastasis remained statistically significant in a multivariate logistic regression analysis, where factors, such as age at diagnosis, primary tumor size, grade, and receptor status, were included as potential confounding variables. No statistically significant correlations were found between the ABCB8 rs4148844 polymorphism and BC phenotype or prognosis. Conclusions and Recommendations Our findings indicate a significant association between the ABCB8 rs17545756 and lymph node involvement in breast cancer patients. Patients carrying the CT genotype and T allele were at a higher risk of lymph node metastasis, suggesting a potential role of this variant in promoting tumor spread. These results suggest that rs17545756 may serve as a potential genetic marker for metastatic risk in breast cancer.

Background and Objective The most prevalent cancer in women worldwide is breast cancer (BC). Research shows that chronic inflammation may be linked to an increased risk of cancer. According to the evidence, the Toll-like receptor (TLR) family plays a significant part in controlling inflammation, innate immunity, cell division, and survival. Among the TLR family, TLR4 is one of the most researched members. It has been noted that BC cells exhibit high mRNA and protein expression levels of TLR4. This implies that single nucleotide polymorphisms (SNPs) in TLR4 gene may have functional effects on the morphology of BC and patient survival. Nevertheless, research on the relationship between SNPs in the TLR4 gene and BC is scarce worldwide. This study aimed to assess the impact of SNPs in the TLR4 gene on the clinical and morphological features of BC as well as the overall survival of BC patients. Material and Method The study population included 170 females with histopathologically confirmed earlystage primary BC (age ranged from 30 to 74 years; median – 46 years). Clinical and tumor pathomorphological data (tumor size, involvement of lymph nodes, metastases, differentiation degree, status of estrogen and progesterone receptors, human epidermal growth factor receptor 2 status, disease progression, and death) were obtained by oncologists. The follow-up period ended on November 30, 2024. Genomic DNA extraction was performed from patients’ blood using a spin column-based isolation method. Genotyping of TLR4 rs7873784 and rs7860896 was conducted by using the TaqMan probe assays on the QuantStudio™ 3 Real-Time PCR System. The strength of potential associations between SNPs and BC features were estimated by crude odds ratio (ORs) and 95% confidence intervals (CIs). Survival curves were generated using the Kaplan-Meier analysis. All statistical tests were performed using IBM SPSS Statistics 30.0.0.0 software, and p<0.05 was considered statistically significant. The study was approved by Kaunas Regional Biomedical Research Ethical Committee (no. BE-2-10 and P1-BE-2-10/2014). Results The genotype frequencies of tagging SNPs were all in agreement with Hardy-Weinberg equilibrium (p>0.05). The genotype distribution of rs7873784 was 0.788 for GG, 0.194 for GC, and 0.018 for CC. For rs7860896 AA, AG and GG genotypes, the distribution was 0.841, 0.147 and 0.012, respectively. The study demonstrated that rs7873784 and rs7860896 were associated with the tumor’s status of progesterone receptors. Females having the rs7873784 GC genotype compared to GG were predisposed to lower rates of progesterone receptor-negative tumors (OR=0.395; 95% CI 0.166-0.938; p=0.035). Meanwhile, rs7860896 AG genotype compared to AA was significantly associated with increased chances of progesterone receptors-negative BC (OR=2.400; 95% CI 1.007-5.718; p=0.048). No additional significant relationships were observed between the SNPs and the studied BC features. Furthermore, the SNPs in TLR4 gene did not correlate with the overall survival of the patients. Conclusions and Recommendations Results suggest that heterozygous genotypes of TLR4 rs7873784 and rs7860896 are associated with early-stage primary BC status of progesterone receptors and may be advantageous genetic biomarkers to assess the prognosis of BC. In order to evaluate the validity of these associations, more extensive and functional research is required.

Introduction Breast cancer (BC) remains the most common oncological disease and a leading cause of cancer death among women worldwide [1]. Studies have shown that single nucleotide polymorphisms (SNPs) can cause alterations in gene expression and play a significant role in various cancers [2]. Since apoptosis is a critical process in BC pathogenesis, the analysis of SNPs in genes involved in this process can provide valuable insights into the mechanisms of BC that are not fully understood [3]. Aim The aim of this study was to analyze SNPs in apoptosis-related genes using whole-exome sequencing (WES) and evaluate the associations of selected SNPs with BC phenotype and prognosis. Methods The cohort consisted of 139 BC patients from the Hospital of Lithuanian University of Health Sciences Kaunas Clinics. For SNPs analysis, genomic DNA was extracted from peripheral blood. WES was performed on 52 randomly selected BC patients. The obtained data were used to analyze polymorphisms in 44 apoptosis-related genes (such as BBCL, TNFRSF, CASP, FASL, etc.) using Franklin platform by Genoox. Missense or UTR variants with a frequency of 5-20% (according to the 1000 Genomes Project) were selected and genotyped in the total study cohort using real-time PCR. Associations between polymorphisms and BC phenotype and prognosis were analyzed. Results In this study only 5 missense variants in apoptosis-related genes with a frequency of 5-20% were identified using WES: TNFRSF10B rs1047266, TNFRSF25 rs11800462, CASP7 rs11555408, CASP8 rs1045485, and BIRC7 rs1077019. Genotyping results of 139 BC patients showed that there were no statistically significant associations between the studied polymorphisms in allelic and genotypic models and tumor pathomorphological features and prognosis, including TNM status, receptor (ER, PR, HER2) status, progression, and death (p>0.05). Conclusions We identified 5 missense polymorphisms in apoptosis-related genes, however, their effect on BC pathogenesis was non-significant.

Introduction Angiosarcoma is a primary malignant tumor of vascular origin, accounting for 1–2% of all soft tissue sarcomas. Its occurrence in the aorta is extremely rare. Epithelioid angiosarcoma is a morphological variant of angiosarcoma, characterized by neoplastic endothelial cells with predominantly epithelioid features. Its incidence is extremely low, and it rarely originates in large arteries [1] . Case Presentation A 55-year-old man presented with complaints of foot pain for several days, burning sensation, itching. Ultrasound showed no blood flow in both legs below the knee. Computed tomography angiography (CTA) revealed that the infrarenal aortic lumen was irregularly thrombosed with extensive parietal thrombi (50%). Stenosis of the left renal artery (90%) and the right popliteal artery (70%) was discovered. A biopsy of aorta was performed. The patient underwent surgery, including resection of the abdominal aorta, aortobifemoral bypass grafting, left renal artery grafting, and thrombectomy of the right and left popliteal arteries. Blood circulation in the left leg was not restored, necessitating left leg amputation. Histological examination confirmed an intravascular epithelioid hemangiosarcoma. The patient was referred to an oncologist and prescribed chemotherapy consisting of 6 cycles of doxorubicin and ifosfamide. No tumor spread was observed on CT. After 2 months, due to obstructive thrombosis of the right popliteal artery, an amputation was performed. The patient was discharged for outpatient treatment. Discussion Primary aortic angiosarcoma is rare, and its preoperative diagnosis is highly challenging, often leading to a poor prognosis. The tumor may mimic an infected aortic aneurysm or a mural thrombus. This type of tumor typically develops without noticeable symptoms, with distal embolism often being the first indication. By the time it is identified, the tumor is usually extensive, and the prognosis remains poor [2,3] . Conclusions Clinical suspicion of angiosarcoma is vital for an early diagnosis and proper treatment.