Gedvilaitė-Vaicechauskienė, Greta

Hipofizės adenomos (HA) yra viena dažniausių pirminių smegenų navikų formų, kurioms būdingas heterogeniškas biologinis elgesys – nuo kliniškai indolentinių iki invazyvių ir linkusių kartotis navikų. Nepaisant pažangos diagnostikoje ir gydyme, išlieka poreikis identifikuoti patikimus molekulinius prognozinius žymenis, leidžiančius tiksliau prognozuoti ligos eigą ir individualizuoti pacientų priežiūrą. Šio darbo tikslas – įvertinti telomerų biologijos, JAK–STAT bei somatostatino signalinių kelių genetinių variantų, jų koduojamų baltymų ir santykinio leukocitų telomerų ilgio sąsajas su hipofizės adenomos pasireiškimu bei naviko biologiniu elgesiu (aktyvumu, invazyvumu, atkryčiu ir dydžiu). Tyrime kompleksiškai analizuoti telomerų homeostazėje dalyvaujantys genai (TERT, TERC, TEP1, TERF1, TERF2, TNKS2, CTC1, ZNF676), imuninės signalizacijos komponentai (STAT4) bei somatostatino receptorių sistema (SSTR2, SSTR5, AIP). Taip pat vertintas santykinis leukocitų telomerų ilgis ir Ki-67 proliferacijos indeksas. Nustatyta, kad TERC rs35073794 genetinis variantas yra reikšmingai susijęs su hipofizės adenomos pasireiškimu ir jos dydžiu. HA pacientų grupėje nustatytas didesnis santykinis leukocitų telomerų ilgis, o atskiri telomerų reguliacijoje dalyvaujančių genų variantai (pvz., TERF1, TNKS2, CTC1) siejasi su santykiniu leukocitų telomerų ilgiu bei ligos pasireiškimo galimybe. Imuninės signalizacijos analizė parodė, kad STAT4 genetiniai variantai ir padidėjusi šio baltymo koncentracija yra susiję su didesne HA pasireiškimo galimybe. Ki-67 proliferacijos indeksas daugumai tiriamųjų buvo mažas, o statistiškai reikšmingų sąsajų su naviko biologinėmis savybėmis ar tirtais genetiniais variantais nenustatyta. Atliktas tyrimas yra pirmasis Lietuvoje, kuriame kompleksiškai įvertintos telomerų biologijos, imuninės signalizacijos ir somatostatino receptorių sistemos genetinės bei molekulinės sąsajos su hipofizės adenoma. Gauti rezultatai papildo esamas žinias apie HA patogenezę ir sudaro prielaidas tolesniems tyrimams, orientuotiems į prognozinių žymenų paiešką bei individualizuotos pacientų priežiūros tobulinimą.

Laryngeal squamous cell carcinoma (LSCC) is a common and aggressive head and neck malignancy with poor prognosis due to late diagnosis and limited biomarkers. Angiogenesis, driven by vascular endothelial growth factor A (VEGFA), plays a key role in tumor growth and metastasis. Single nucleotide variants (SNVs) in the VEGFA gene have been linked to various cancers, but their role in LSCC remains unclear.

Purpose: This study aims to evaluate TERC and TERT gene polymorphisms and their relationship to risk factors, laboratory findings, and imaging in endocrine ophthalmopathy (EO) patients. Methods: This cross-sectional study involved EO patients and healthy volunteers who were patients of the Eye Disease Clinic of the Lithuanian University of Health Sciences. A complete ophthalmological examination was performed on all participants. The results of imaging and laboratory tests were taken from the medical records. DNA was extracted from peripheral blood leukocytes, and real-time polymerase chain reaction (RT-PCR) was used to analyze the TERC rs35073794 and rs12696304, and TERT rs2736098 gene polymorphisms. Data analysis was performed using IBM SPSS Statistics 27 software. Results: According to the RT-PCR results, the TERC rs35073794 GG genotype polymorphism is significantly associated with EO (p=0.001). In addition, the TERC rs12696304 polymorphism CC genotype correlates with extraocular muscle changes observed in imaging studies (p=0.017). In contrast, the TERT rs2736098 polymorphism CC genotype is associated with increased anti-TSHR levels (p=0.047), and the CT genotype is associated with smoking (p=0.035). Conclusions: This study provides new evidence that the TERC rs35073794 polymorphism is related to EO, the TERC rs12696304 polymorphism correlates with extraocular muscle changes, and the TERT rs2736098 polymorphism is associated with increased anti-TSHR levels and smoking. These results suggest that TERC and TERT gene polymorphisms have the potential to be used as prognostic factors in clinical practice. However, larger multicenter studies still need to validate these results before they can be introduced into clinical practice.

Purpose: To investigate whether the single-nucleotide polymorphisms (SNP) and haplotypes of the SIRT1(rs3818292, rs3758391, rs7895833) are associated with optic neuritis (ON) and multiple sclerosis (MS). Methods: We studied: a control group, patients with MS, with ON, and with ON and MS. DNA was extracted from peripheral blood leukocytes and genotyped by rPCR reaction. The data of the SNP of the SIRT1 gene rs3818292, rs3758391, and rs7895833 were then analyzed. Results: rs3818292 is associated with a 2.19-fold increased odd of ON occurrence by the best-fit overdominant model (p=0.002). rs3758391 is associated with 2.16-fold increased odds of ON occurrence under the best-fit dominant model (p=0.004). rs7895833 is associated with a 2.38-fold increased odd of ON occurrence under the best-fit overdominant model (p=0.002). rs3818292 is associated with a 2.14-fold increased odds of MS occurrence under the bestfit dominant model (p=0.015). rs7895833 is associated with a 2.31-fold increased odds of MS occurrence by the best-fit overdominant model (p=0.002). rs3818292 is associated with 2.91-fold increased odds of ON with MS occurrence by the best-fit overdominant model (p=0.018). rs3758391 is associated with 2.55-fold increased odds of ON with MS occurrence under the best-fit dominant model (p=0.015). rs7895833 is associated with 3.04-fold increased odds of ON with MS occurrence under the best-fit overdominant model (p=0.005). Individuals carrying rs3818292-rs3758391-rs7895833 haplotype A-T-A is likely to be associated with 2.3-fold decreased odds of MS occurrence (p=0.031), and G-T-G haplotype is associated with 1.7-fold increased odds of MS occurrence (p=0.047). Individuals carrying rs3818292-rs3758391- rs7895833 haplotype G-T-G is likely to be associated with 2-fold increased odds of ON occurrence (p=0.038). Conclusions: SIRT1 rs3818292, rs3758391, rs7895833 SNP, and haplotypes are statistically significantly associated with ON and MS development.

Introduction Pituitary adenomas (PAs) are benign tumors of the pituitary gland, a small structure at the brain’s base responsible for hormone regulation [1]. Although typically slow-growing and non-metastatic, PAs can cause complications if they enlarge and press on surrounding brain structures [2]. Growing evidence suggests that genetic factors contribute to PA development, with Signal Transducer and Activator of Transcription 4 (STAT4) emerging as a key focus. STAT4 regulates gene expression and modulates immune responses [3], and its dysregulation or overactivation may promote inflammation, potentially influencing oncogenesis. This study explores STAT4’s role in PA pathogenesis, uncovering potential links that could inform future therapies and deepen our understanding of the disease. Aim This study aims to investigate whether STAT4 contributes to PA recurrence by examining STAT4 polymorphisms and serum levels. Methods Examining 139 patients with PA and 357 healthy subjects, DNA samples from peripheral blood leukocytes were purified using the DNA salting-out method. Real-time polymerase chain reaction (RT-PCR) assessed STAT4 (rs7574865, rs10181656, rs7601754, and rs10168266) single nucleotide polymorphism (SNPs), and enzyme-linked immunosorbent assay (ELISA) determined the levels of STAT4 in blood serum. The analysis was conducted using “IBM SPSS Statistics 29.0“. Results For PA recurrence, the rs7574865 G allele was less frequent in PA without recurrence (p = 0.012), and the GT+GG genotypes were associated with a 1.8-fold increase in PA without recurrence (p = 0.008). Serum STAT4 levels were higher in PA patients compared to the reference group (p < 0.001). Elevated STAT4 serum levels were observed in PA patients with the STAT4 rs10181656 CC or CG genotypes (CC: p = 0.004; CG: p = 0.023) and with the rs7574865 GG or GT genotypes (GG: p = 0.003; GT: p = 0.021). PA patients with the STAT4 rs7601754 AA genotype exhibited higher serum levels compared to the reference group (p < 0.001). Similarly, higher serum levels were found in PA patients with STAT4 rs10168266 CC or CT genotypes (CC: p = 0.004; CT: p = 0.027). Conclusions STAT4 genetic variations and serum levels are linked to PA recurrence. The rs7574865 G allele was less frequent in non-recurrent PA, suggesting a protective effect. Also, PA patients had higher STAT4 levels. These findings highlight STAT4’s potential as a biomarker for PA recurrence and targeted therapy. Further research is needed to explore its potential as a target for personalized treatment strategies.

Background. Pituitary adenoma (PA) is one of the most common brain tumours, yet its pathogenesis remains unclear [1]. PA varies in invasiveness; some remain confined to the pituitary gland, while others aggressively infiltrate surrounding tissues, leading to more severe clinical complications [2]. Molecular biomarkers indicate biological and pathological processes or therapeutic responses, offering valuable insights for diagnosis and prognosis [3]. CXCL12, a homeostatic chemokine, induces migration and activation of haematopoietic progenitors, endothelial cells, and leukocytes, regulating the tumour microenvironment by promoting angiogenesis and immune cell recruitment, potentially aiding tumour growth [4]. Polymorphisms within the CXCL12 gene, particularly those influencing its expression or function, may affect PA susceptibility, tumour size, invasiveness, and response to treatment [5]. A deeper understanding of the genetic mechanisms underlying CXCL12 may contribute to developing personalised therapeutic approaches and enhancing prognostic tools for PA management. Aim. The current study aimed to investigate the associations between the CXCL12 rs1801157, rs2297630 polymorphisms, and the PA invasiveness in females. Methods. A case-control study enrolled 90 females with PA and 160 healthy females. DNA samples from peripheral blood leukocytes were purified by the DNA salting-out method. Single nucleotide polymorphisms (rs1801157, rs2297630) were determined using real-time polymerase chain reaction (RT-PCR). The results were analysed using the IBM SPSS Statistics 29.1 statistical analysis method. Results. We found that CXCL12 rs1801157 TT genotype and T allele were statistically significantly more frequent in the non-invasive PA group compared to control group females (8.7 vs. 2.5, p = 0.043, 28.3 vs 16.9, p = 0.014, respectively). In the invasive PA group, it was found that CXCL12 rs2297630 AA genotype and A allele were statistically significantly more frequent in the control group than in the invasive PA group females (8.1 vs. 0, p = 0.009, 29.7 vs. 13.6, p = 0.002, respectively). A binary logistic regression analysis evaluated the impact of CXCL12 rs1801157 and rs2297630 on non-invasive/invasive PA development. The analysis revealed that CXCL12 rs1801157 T allele increased the odds of non-invasive PA’s development 1.9.-fold under the additive model (OR: 1.951, CI: 1.124–3.386, p = 0.018). Moreover, the CXCL12 rs2297630 A allele decreases the odds of invasive PA development 2.8-fold under the additive model (OR: 0.359, CI: 0.183–0.705, p = 0.003). Conclusions. The CXCL12 rs2297630 A allele suggests a potential protective effect against invasive PA development. In contrast, the CXCL12 rs1801157 T allele may be a risk factor for developing non-invasive forms of PA.

Background. Multiple sclerosis (MS) is an autoimmune disorder that affects the central nervous system and is characterised by gliosis, demyelination, inflammation, and degeneration of nerve cells [1]. Young adults with MS, usually between the ages of 20 and 30, often present with unilateral optic neuritis, partial myelitis, sensory abnormalities, or brainstem syndromes [2]. MS and Signal Transducer and Activator of Transcription 4 (STAT4) may be interconnected, as MS has been associated with immune system dysfunction [3]. Janus kinases (JAKs) are the proteins that enable members of class I and class II cytokine receptor families to transmit their signals. Once activated, JAKs phosphorylate the STATs. After phosphorylation, the STAT proteins undergo cytoplasmic dimerization before translocating to the nucleus, where they bind to deoxyribonucleic acid (DNA) regulatory elements and initiate gene transcription. Consequently, variations in STAT4 expression or activity may influence the immune system’s normal response and function, potentially resulting in immunosuppression or autoimmune disorders [4]. Overall, the connection between the STAT4 gene and MS suggests that dysregulation of the immune system plays a significant role in the development of the disease [3]. Aim. This study aimed to evaluate the association between the STAT4 rs7601754 polymorphism and vision impairment in individuals with MS. Methods. Altogether, 400 patients were enrolled in the study (MS patients with vision impairment (n = 131), MS patients without vision impairment (n = 69), and healthy controls as a reference group (n = 200)). Blood samples were collected, and the DNA salting-out method was used for DNA extraction from peripheral venous blood. Genotyping of STAT4 rs7601754 was performed using RT-PCR. Results. We found that the STAT4 rs7601754 A allele was statistically significantly less frequent in MS patients with and without vision impairment compared to the reference group (p = 0.046, p < 0.001, respectively). Binary logistic regression analysis revealed that the AG + GG genotypes are associated with 2.7-fold increased odds of not having visual impairment in MS patients (OR: 2.656; CI: 1.493–4.724; p < 0.001). Conclusion. This study aimed to enhance the understanding of its role in the development of vision-related symptoms in MS patients, potentially revealing a biomarker for vision impairment in MS. STAT4 rs7601754 AG + GG genotypes are linked to a 2.7-fold increase in the odds of not experiencing visual impairment in MS patients.

Background and Objectives: Age-related macular degeneration (AMD) is the leading cause of blindness, affecting millions worldwide. Its pathogenesis involves the death of the retinal pigment epithelium (RPE), followed by photoreceptor degeneration. Although AMD is multifactorial, various genetic markers are strongly associated with the disease and may serve as biomarkers for evaluating treatment efficacy. This study investigates TAB2 rs237025, IKBKB rs13278372, and IKBKG rs2472395 variants and their respective serum protein concentrations in relation to AMD occurrence and exudative AMD treatment response to anti-VEGF treatment. Materials and Methods: The case–control study involved 961 individuals, and they were divided into three groups: control, early AMD, and exudative AM patients. Genotyping of selected SNPs were conducted using a real-time polymerase chain reaction method (RT-PCR). Based on the clinical OCT and BCVA data, patients with exudative AMD were categorized into one of two groups: responders and non-responders. The data obtained were analyzed using the “IBM SPSS Statistics 29.0” software program. Results: Our study revealed that TAB2 rs237025 allele A was identified as a risk factor for early and exudative AMD development. The same associations remained only in females with exudative AMD but not in males, suggesting gender-specific pathogenetic pathways in exudative AMD. Analysis of IKBKB rs13278372 or serum IKBKB protein associations with early or exudative AMD occurrence in the Lithuanian population revealed no significant associations. On the other hand, we found that each A allele of IKBKB rs13278372 was associated with a worse response to anti-VEGF treatment (OR = 0.347; 95% CI: 0.145–0.961; p = 0.041). These results suggest a potential marker for future studies evaluating anti-VEGF treatment for exudative AMD patients. IKBKG rs2472395 was a protective variant for early AMD in males and for exudative AMD in females only. Also, IKBKG protein concentration was lower in exudative AMD relative to the control group (median (IQR): 0.442 (0.152) vs. 0.538 (0.337), p = 0.015). Moreover, exudative AMD patients who carry the GG genotype of IKBKG rs2472394 exhibited significantly reduced serum IKBKG concentrations compared to the controls (median (IQR): 0.434 (0.199) vs. 0.603 (0.335), p = 0.012), leading to the hypothesis that the IKBKG rs2472394 variant might play a role in protein concentration differences and exudative AMD development. Conclusions: Our study identified the TAB2 rs237025 allele A as a significant risk factor for both early and exudative AMD, with gender-specific associations observed in females with exudative AMD, suggesting distinct pathogenetic pathways. While IKBKB rs13278372 and serum IKBKB protein levels showed no significant association with AMD development, the A allele of IKBKB rs13278372 was associated with a worse response to anti-VEGF treatment, indicating its potential as a marker for treatment outcomes. Additionally, the IKBKG rs2472395 variant was found to be protective for early AMD in males and exudative AMD in females, and lower IKBKG protein levels were associated with exudative AMD, particularly in patients with the GG genotype of IKBKG rs2472394, suggesting its role in protein concentration and disease progression. These findings highlight genetic markers that may contribute to AMD pathogenesis and treatment response.

Introduction: Pituitary adenoma (PA) is one of the most common brain tumors, yet its pathogenesis remains unclear. PA varies significantly in activity; some secrete excess hormones, causing symptoms related to hormonal imbalance, while others are nonsecreting but may still cause symptoms due to size, exerting pressure on nearby structures. CXCL12, a homeostatic chemokine regulates the tumor microenvironment by promoting angiogenesis and recruiting immune cells, potentially supporting tumor growth. CXCL12 gene polymorphisms, particularly those affecting expression or function, may influence PA risk, size, invasiveness, and treatment response. Understanding the genetic role of CXCL12 could support personalized treatments and improve prognostic tools for PA management. Materials and Methods: A case-control study enrolled 90 females with PA and 160 healthy females. DNA samples from peripheral blood leukocytes were purified by the DNA saltingout method. Single nucleotide polymorphisms (rs1801157, rs2297630) were determined using real-time polymerase chain reaction (RT-PCR). The results were analyzed using the “IBM SPSS Statistics 29.1“ statistical analysis method. Results: We found that CXCL12 rs2297630 AA genotype and A allele were statistically significantly more frequent in controls compared to the hormonally inactive PAs (8.1 vs. 0, p=0.013, 29.7 vs. 11.7, p=0.003, respectively). In the active PA group, the CXCL12 rs1801157 TT genotype and T allele were statistically significantly more frequent than in the controls (6.7 vs. 2.5, p=0.042, 27.5 vs. 16.9, p=0.012, respectively). However, the rs2297630 A allele was statistically significantly less frequent in the hormonally active PAs than in the controls (19.2 vs. 29.7, p=0.026). CXCL12 rs1801157 T allele was associated with 1.2-fold increased odds of active PA occurrence under the additive model (OR: 1.910, CI: 1.143–3.191, p = 0.014). Furthermore, the CXCL12 rs2297630 A allele decreased the odds of inactive and active PA development by 3.3-fold and 1.8-fold under the additive models (OR: 0.301, CI: 0.130–0.700, p = 0.005; OR: 0.545, CI: 0.320–0.927, p = 0.025, respectively). Conclusions: The CXCL12 rs2297630 A allele suggests potential protective effect against both inactive and active PA development. In contrast, the CXCL12 rs1801157 T allele may be a risk factor for developing more active forms of PA.

Introduction: Pituitary adenomas (PAs) are benign tumors that develop in the pituitary gland, a small but crucial organ located at the base of the brain responsible for regulating hormone production (Lake et al., 2013). Although PAs are generally slow-growing and nonmetastatic, they can cause significant issues if they enlarge and exert pressure on nearby brain structures (Møller et al., 2019). In light of the potential influence of genetic factors on PA development, the Signal Transducer and Activator of Transcription 4 (STAT4) gene has gained attention as a key area of research. This study examines the possible associations between STAT4 polymorphisms—rs7574865, rs10181656, rs7601754, rs10168266 and serum levels with PA occurrence. The goal is to investigate the role of STAT4 in the antitumor immune response and its potential involvement in the initiation of PA. Materials and Methods: This study was conducted at the Laboratory of Ophthalmology, Lithuanian University of Health Sciences, using DNA extracted from peripheral venous blood samples. Genotyping for four STAT4 single nucleotide polymorphisms (SNPs) was performed through real-time PCR with TaqMan® assays. Results: The study found that individuals carrying the STAT4 rs7574865 GT + GG genotypes had a 1.7-fold increased odds of developing PAs under the dominant genetic model (p = 0.012). While no significant associations were observed in females; males with the rs10168266 CC + CT genotypes had 2.5-fold higher odds of PA occurrence than those with the TT genotype (p = 0.005). Also, serum STAT4 levels were higher in PA patients compared to the reference group (p < 0.001). Conclusion: The STAT4 rs7574865 and rs10168266 show significant associations with increased odds of PA occurrence. Elevated serum STAT4 levels were also observed in PA patients, highlighting its potential role in PA pathogenesis.