Receptor Beta Research Articles

A Game Changer for Resistant Hypertension: The Rise of Aprocitentan.

Hypertension is a major risk factor for cardiovascular disease (CVD), and a major contributor to global morbidity and mortality. In particular, resistant hypertension (rHTN), defined as blood pressure that remains elevated despite treatment with at least three antihypertensive agents including a diuretic, continues to be a major pharmacotherapeutic challenge. Traditional antihypertensive drugs often fail in patients with rHTN, underscoring the need for novel therapies. This is a brief mini-review of aprocitentan, a new drug that promises a glimmer of hope for rHTN patients. This drug is a dual endothelin (ET) receptor antagonist that blocks both ETA and ETB receptors. Given that these two receptors are critical players in vasotone regulation, antagonizing them, such as by aprocitentan, would be expected to significantly reduce blood pressure in patients with rHTN. Indeed, the PRECISION clinical trial demonstrated aprocitentan's superior effectiveness in reducing blood pressure in resistant patients, and the effects were sustained. Aprocitentan has been recently FDA-approved, marking a major milestone in hypertension management, offering hope for patients with difficult-to-treat hypertension.

C6orf120 Deficiency Inhibits Hepatic Stellate Cell Activation by Upregulating RARβ Signaling.

Vitamin A (VA) and its active form, retinoic acid (RA), are crucial for preserving hepatic stellate cells (HSCs) quiescence and reversing fibrosis. While C6orf120 is known to be involved in HSC activation, its role in RA signaling is unclear. This study found that C6orf120 knockdown markedly reduced CCL4-induced liver fibrosis and TGF-β1-induced activation in LX-2 cells, a human HSC line. This inhibition was associated with enhanced RA signaling, particularly affecting the RA receptor beta (RARβ). Inhibition of RARβ significantly reversed the protective effects of C6orf120 knockdown, indicating that RARβ signaling contributes to the inhibitory effect of C6orf120 knockdown on HSC activation. Our results reveal that C6orf120 inhibition could be a therapeutic strategy for liver fibrosis by regulating RARβ signaling.

Sodium valproate reverses aortic hypercontractility in acute myocardial infarction in rabbits.

Sympathetic nervous system (SNS), endothelin 1 (ET-1) and angiotensin II (Ang II) are involved in the pathophysiology of acute myocardial infarction (AMI). Valproic acid (VPA) is under study for the treatment against AMI due to its beneficial cardiac effects. However, the vascular effects of VPA on the activation of the SNS, ET-1 and Ang II after AMI are not fully studied. In our study, we used aorta from New Zealand White rabbits without AMI, with AMI and AMI treated with VPA (500mg/kg/day). AMI was induced by occluding the left circumflex coronary artery for 1h, followed by reperfusion. After 5 weeks, we studied the ex vivo vascular reactivity in organ bath and measured protein expression by Western blot. Our findings indicated that AMI increased vasoconstriction to exogenous and endogenous NE and ET-1, which was reversed by VPA eliciting upregulation of α2-adrenergic and ETB receptors and downregulating ETA receptors. Although no changes in the vascular response to Ang II were observed in AMI or VPA-treated rabbits, an increase in Ang II type 2 receptor expression was found in VPA-treated rabbits. We conclude that VPA could be considered a vascular protector by modulating SNS, ET-1 and Ang II in the aorta, which together with its cardioprotective effect would make it a promising candidate for the treatment of AMI.

Association between estrogen receptors polymorphisms and temporomandibular joint disorders: A systematic review.

This systematic review aimed to investigate the association between genetic polymorphisms in estrogen receptor alpha (ERα) and beta (ERβ) and the presence and severity of temporomandibular disorders (TMDs). A comprehensive search was conducted in PubMed, Scopus and Web of Science on October 19, 2024, without language or time restrictions. Studies were included if they were cross-sectional, case-control, or cohort designs and reported data on ER-polymorphisms and TMDs, diagnosed using validated criteria. Two independent reviewers screened the results to identify eligible studies. Data were extracted and synthesized narratively due to high heterogeneity across included studies, with a subgroup analysis performed to evaluate the influence of sex. The risk of bias was assessed using the Joanna Briggs Institute's critical appraisal tools. The search identified 277 articles, of which 8 studies met the inclusion criteria. Seven were case-control and two cross-sectional studies. The most commonly investigated ER polymorphisms were ESR1 (Pvu II and Xba I) and ESR2 (rs1676303). Some studies identified associations between specific polymorphisms and TMD-related pain or joint conditions, although results varied across different populations and subtypes of TMDs. The findings suggest potential associations between specific ERα and ERβ polymorphisms and TMD risk, particularly in women. However, inconsistencies across studies underscore the need for prospective and larger studies to confirm these genetic links and clarify their clinical significance. CRD42024581266.

Neuroprotective role of mirabegron: Targeting beta-3 adrenergic receptors to alleviate ulcerative colitis-associated cognitive impairment.

While cognitive impairment has been documented in ulcerative colitic patients, the possible influence of central β3-adrenergic receptor (β3-AR) signaling on this extraintestinal manifestation remains unclear. Previously, we identified an imperative role for mirabegron (MA) as an agonist of β3-AR, in decreasing the BACE-1/beta-amyloid (Aβ) cue in the colons of UC rats. Consequently, we investigated its therapeutic potential for alleviating cognitive impairment associated with UC. To fulfil our aim, rats administered iodoacetamide were treated with the β3-AR agonist (MA) alone, with the antagonist (SR59230A) for 8 days, or kept untreated. The animals' behavior (MWM and NOR tests) and hippocampal structure were assessed. Mechanistically, necroptosis, ER stress (ERS), Aβ-amyloidosis, inflammation/oxidative burden, and gut/BBB dysfunction were analyzed. Post-administration of MA improved weight gain, colon/hippocampal structures, and memory. Additionally, it inhibited serum levels of lipopolysaccharide and Annexin-1, indicating recovered gut and BBB integrity. MA turned off the pathogenic BACE-1/Aβ axis in the hippocampus, necroptosis trajectory (TNFR-1/RIPK1/RIPK3/MLKL), and the IRE-1α/JNK signal. Moreover, MA enhanced the transcription factor PPAR-γ, decreased NF-κΒ/TNF-α inflammatory hub, and modulated the redox imbalance by decreasing malondialdehyde and increasing catalase. Notably, MA's behavioral, structural, and molecular beneficial actions were hindered by the pre-administration of SR59230A. From a novel standpoint, we recognized the β3-AR as a therapeutic target for UC-associated cognitive impairment in the hippocampus. In this context, the aptitude of MA to inhibit UC-induced hippocampal amyloidogenesis, alongside its anti-necroptotic, anti-ERS, anti-inflammatory, and antioxidant effects, contribute to these central enhancements, while also regulating permeability in both gut and BBB barriers.

A germline PDGFRB splice site variant associated with infantile myofibromatosis and resistance to imatinib

Infantile myofibromatosis is characterized by the development of myofibroblastic tumors in young children. In most cases, the disease is caused by somatic gain-of-function variants in platelet-derived growth factor (PDGF) receptor beta (PDGFRB). Here, we reported a novel germline intronic PDGFRB variant, c.2905-8G>A, in 6 unrelated infants with multifocal myofibromatosis and their relatives. We performed constitutional and tumor DNA and RNA sequencing to identify novel variants, which were subsequently characterized in cellular assays. All patients had multiple skin nodules, 4 had bone lesions, and 2 had aggressive disease with bowel obstruction. The c.2905-8G>A substitution creates an alternative acceptor splice site in intron 21, inserting 2 codons in the PDGFRB transcript. Functional studies revealed that the splice change induced a partial loss of function, contrasting with previously described variants. In 4 tumor samples, we identified a second somatic hit at position Asp850 in PDGFRB exon 18, triggering constitutive receptor activation and resistance to imatinib. In addition to vinblastine and methotrexate, 2 patients received imatinib without objective response. One of them switched to dasatinib with concomitant improvement. This splice-site PDGFRB variant favors the development of myofibroma, featuring an acquired oncogenic variant in the same gene and resistance to targeted therapy.

Asymmetric synthesis of an optically active 1-benzylindane derivative as a selective agonist for estrogen receptor beta

1-(3-Fluoro-4-hydroxybenzyl)-5-hydroxy-2,3-dihydro-1H-indene-1-carbonitrile rac-1a is a potent and highly selective agonist for estrogen receptor β that shows potential as a therapeutic agent for obesity and depression in postmenopausal women. To assess the more precise pharmacological actions of rac-1a, a stable and large-scale supply of optically active (R)-1a is required. In this study, our objective was to synthesize (R)-1a by developing a rapid and more efficient asymmetric synthetic pathway. The rapid supply of (R)-1a was accomplished by diastereomeric resolution of the easily obtainable racemic precursor 1-(3-fluoro-4-methoxybenzyl)-5-methoxy-2,3-dihydro-1H-indene-1-carboxylic acid rac-3a using a chiral oxazolidinone auxiliary. Despite the need to discard the unwanted enantiomer, this approach enabled stable gram-scale supply of (R)-1a by a short process that minimized purification by column chromatography. Furthermore, by exploring more efficient synthetic methods, we synthesized chiral precursor (R)-3a by asymmetric desymmetrization of 1,3-diol 5, which is an important reaction in asymmetric synthesis.

Interaction between Estrogen Receptors and p53: A Broader Role for Tamoxifen?

Tamoxifen is one of the most widely used anticancer drugs in the world. It is a safe drug with generally well-tolerated side effects and has been prescribed for the treatment of early-stage and advanced-stage or metastatic estrogen receptor alpha (ERα/ESR1) -positive breast cancer. Tamoxifen therapy also provides 38% reduction of the risk of developing breast cancer in women at high risk. With the advent of newer medications targeting ERα-positive breast cancer, tamoxifen is now mainly used as adjuvant therapy for lower-risk premenopausal breast cancer and cancer prevention. It is widely accepted that tamoxifen as a selective estrogen receptor modulator (SERM) exerts its therapeutic effect by competitively binding to ERα leading to recruitment of co-repressors and inhibition of transcription of genes involved in the proliferation of breast cancer epithelium. As such, expression of ERα in breast tumors has been considered necessary for tumors to be responsive to tamoxifen therapy. However, ERα-independent effects of tamoxifen in various in vitro and in vivo contexts have been reported over the years. Importantly, recent discovery that ERα and estrogen receptor beta (ERβ/ESR2) can bind tumor suppressor protein p53 with functional consequences has provided new insights into the mechanisms underlying response to tamoxifen therapy and resistance. Furthermore, these findings have paved the way for broadening the use of tamoxifen by potentially repurposing it to treat triple negative (negative for ERα, human epidermal growth factor receptor 2/HER2, and progesterone receptor/PR) breast cancer (TNBC). Herein, we summarize these developments and discuss their mechanistic underpinnings and clinical implications.

Nucleus pulposus cell network modelling in the intervertebral disc

Intervertebral disc degeneration (IDD) results from an imbalance between anabolic and catabolic processes in the extracellular matrix (ECM). Due to complex biochemical interactions, a comprehensive understanding is needed. This study presents a regulatory network model (RNM) for nucleus pulposus cells (NPC), representing normal intervertebral disc (IVD) conditions. The RNM includes 33 proteins, and 153 interactions based on literature, incorporating key NPC regulatory mechanisms. A semi-quantitative approach calculates the basal steady state, accurately reflecting normal NPC activity. Model validation through published studies replicated pro-catabolic and pro-anabolic shifts, emphasizing the roles of transforming growth factor beta (TGF-β) and interleukin-1 receptor antagonist (IL-1Ra) in ECM regulation. This IVD RNM is a valuable tool for predicting IDD progression, offering insights into ECM degradation mechanisms and guiding experimental research on IVD health and degeneration.

In Vivo Effects of Cardiomyocyte-Specific Beta-1 Blockade on Afterload- and Frequency-dependent Cardiac Performance.

Pharmacologic beta-blockade is a well-established therapy for reducing adverse effects from sympathetic overactivity in cardiovascular diseases, such as heart failure. Despite decades of research efforts, in vivo cardiac functional studies utilizing genetic animal models remain scant. We generated a mouse model of cardiomyocyte-specific deletion of beta-1 adrenergic receptor (ADRB1) , the primary subtype expressed in cardiac myocytes, and demonstrated the role for ADRB1 in the maintenance of cardiac function at baseline and during exposure to increase in cardiac afterload by transient aortic occlusion and increasing heart rates (HRs) via atrial pacing. cKO hearts showed mildly depressed baseline left ventricular (LV) function, including slower HR, decreased contractility (dP/dtmax/IP) and prolonged relaxation (Tau) at baseline in both sexes. Exposure to increased LV afterload depressed LV function in either genotype similarly; however, the functional recovery following the removal of the afterload was severely impaired in cKO hearts, while cardiac function was immediately normalized in WT hearts. When HR was altered from 400 to 700bpm, cKO hearts were deficient in HR-dependent improvement of cardiac contractility and relaxation, known as positive force frequency relationship, that was evident in WT hearts. Enhanced phosphorylation of phospholamban by the HR increase was markedly blunted in cKO myocardium vs wild types, while CaMKII phosphorylation was comparable between the genotypes, suggesting the critical involvement of PKA. These results provide the first experimental evidence for the role of ADRB1 in cardiomyocytes for maintaining cardiac function at baseline and during acute stress, providing clinical perspective relating to the management of patients on beta-blockers.

Anti-retinal immune response in sarcoid uveitis: A potential role for PCLO as an antigenic target.

To explore the autoimmune component of sarcoid uveitis (SU) by analyzing serum anti-retinal antibodies (ARAs), identifying targeted retinal proteins, T- and B-cell receptor repertoires and HLA genotype. Material from 45 sarcoidosis patients with no presenting uveitis (SNPU) and 46 with SU was analyzed. Serum ARAs and targeted retinal layers were assessed using indirect immunofluorescence staining. HuScan analysis identified autoantibody-targeted linear epitopes. Validation included a bead-based assay for anti-Piccolo Presynaptic Cytomatrix Protein (PCLO) antibodies and an ELISpot assay for PCLO-reactive T-lymphocytes. T cell receptor beta (TCRB) and B cell receptor heavy (BCRH) repertoire analyses were performed using next-generation sequencing and HLA class II genotypes were determined by sequence-specific primer analysis. ARAs were more prevalent in SU patients than in SNPU patients (52 vs. 22%, p=0.003), with significant more reactivity against the nuclear retinal layer (32 vs. 7%, p=0.005). HuScan identified autoantibodies against three retinal proteins, including PCLO. Bead-based analysis showed higher anti-PCLO autoantibody levels in ARA-positive patients (median: 913.3 vs. 544.5, p=0.035), and PCLO-directed T-lymphocytes were present in ARA-positive SU patients. Two TCRB clusters were identified in four unique ARA positive patients, while absent in ARA negative patients. No HLA allele association with ARA status could be detected. Our findings reveal an association between serum ARA-positivity and SU, suggesting a link to autoimmune processes. An humoral and cellular response against the retinal protein PCLO was identified, highlighting PCLO as a potential autoimmune target in ARA-positive patients. Additionally, specific TCRB clusters were found to correlate with ARA status.

Scrutinizing the evidence of anthracene toxicity on adrenergic receptor beta-2 and its bioremediation by fungal manganese peroxidase via in silico approaches.

Exposure to anthracene can cause skin and eye irritation, respiratory issues, and potential long-term health risks, including carcinogenic effects. It is also toxic to aquatic and human life and has the potential for long-term environmental contamination. This study aims to alleviate the adverse environmental effects of anthracene through fungal degradation, focusing on bioremediation approaches using bioinformatics. Toxicity prediction using Pro-Tox 3.0 identified anthracene as a compound of toxicity class 4 with a LD50 of 316mg/kg. Sequence of manganese peroxidase from Lachnellula suecica and human adrenergic receptor beta 2 were retrieved from NCBI databases. Secondary structure analysis using SOPMA indicated that both manganese peroxidase and adrenergic receptor beta 2 contain significant random coil content (56.57% and 51.57% respectively) followed by alpha-helix and beta-turns. The tertiary structure of both proteins was predicted using the SWISSMODEL tool and molecular docking using Autodock vina revealed strong binding affinities of anthracene with adrenergic receptor beta 2, showing a binding energy of - 6.6kcal/mol with anthracene confirming the negative impacts on human health. To mitigate the anthracene pollution, further docking indicated Anthracene-2,6-dicarboxylic acid as the most vigorous ligand for manganese peroxidase of L. suecica with a binding energy of - 9.3kcal/mol, suggesting its potential as a bioremediating agent. Visualization using Discovery Studio elucidated the molecular interactions within the docked complex. Molecular dynamics simulations using the OpenMM engine and AMBER force field confirmed stable enzyme-ligand complexes, highlighting the potential of manganese peroxidase for sustained enzymatic activity against anthracene.

The synergistic effects of citicoline and silymarin on liver injury and thyroid hormone disturbances in γ-irradiated rats.

Exposure to ionizing radiation is inevitable due to its extensive use in industrial and medical applications. The search for effective and safe natural therapeutic agents as alternatives to chemical drugs is crucial to mitigate their side effects. This study aimed to evaluate the effects of citicoline as a standalone treatment or in combination with the anti-hepatotoxic drug silymarin in protecting against liver injury caused by γ-radiation in rats. The rats were exposed to γ-radiation (7Gy) and treated with citicoline (300mg/kg/day) and/or silymarin (50mg/kg/day). The results showed that citicoline alleviated liver damage in irradiated rats by reducing hepatic malondialdehyde levels, serum aspartate aminotransferase activity, and inflammatory mediators such as tumor necrosis factor-alpha (TNF-α), interleukin-1 beta (IL-1β), and nuclear factor-kappa B (NF-κB). It also increased acetylcholine (ACh) levels and the gene expression of the anti-inflammatory protein α7 nicotinic acetylcholine receptor (α7nAChR). Additionally, citicoline improved serum triiodothyronine (T3) levels, thyroid hormone receptor beta (TRβ) gene expression, and iodothyronine deiodinase type 1 activity in hepatic tissues of irradiated rats. Furthermore, citicoline enhanced the effects of silymarin on thyroxine (T4), TRβ, ACh, and α7nAChR when co-administered in irradiated rats. Histopathological analysis confirmed these findings, demonstrating improved liver tissue structure. Citicoline mitigates γ-radiation-induced liver damage by reducing oxidative stress, activating the cholinergic anti-inflammatory pathway, and modulating thyroid hormone metabolism. These findings support the use of citicoline as a safe standalone treatment or as an adjuvant with silymarin for managing liver damage and thyroid hormone disturbances caused by γ-irradiation.

Polymorphisms and dental age in non-syndromic cleft lip and palate: a cross-sectional study.

Children with non-syndromic cleft lip with or without palate (CL ± P) may present alterations in dental development. The purpose of this cross-sectional study was to compare the dental age (DA) between children with and without CL ± P, and whether single nucleotide polymorphisms (SNPs) in genes encoding growth factors are associated with DA variations. Children aged between 5 and 14 years with and without CL ± P were recruited to participate in this study. DA was evaluated by calibrated examiners (kappa > 0.80) using the method proposed by Demirjian et al. (1973). Genomic DNA was extracted from buccal cells, and SNPs in Epidermal Growth Factor (EGF) - rs4444903 and rs2237051, Epidermal Growth Factor Receptor (EGFR) - rs2227983 -, Transforming Growth Factor Beta 1 (TGFB1) - rs1800470 and rs4803455 -, and Transforming Growth Factor Beta Receptor 2 (TGFBR2) - rs3087465 - were genotyped by real-time polymerase chain reactions using the TaqMan assay. The Student T-test was used to compare the variations in DA between the phenotypes "with CL ± P" and "without CL ± P", and the ANOVA two-way test was performed to compare the variations in DA among the genotypes (α = 0.05). A post-hoc analysis was performed using Bonferroni correction. Two hundred and nine (n = 209) children (100 with CL ± P and 109 without CL ± P) with a mean chronological age of 8.66 years - standard deviation (SD) = 1.92 - were included. The group with CL ± P demonstrated a significantly delayed DA (mean=-0.23; SD = 0.71) compared to the group without CL ± P (mean=-0.01; SD = 0.88) (p = 0.049). Genotype distributions were in Hardy-Weinberg equilibrium. The SNP rs4803455 in TGFB1 was significantly associated with DA variations in children without CL ± P (p < 0.01). In the group with CL ± P, no significant differences in DA were observed among the genotypes. Children with CL ± P presented delayed DA compared with children without CL ± P. The SNP rs4803455 in TGFB1 is associated with variations in DA in children without CL ± P.

P-63 PREVENTION OF POSSIBLE ABORTIONS IN PREGNANT WOMEN WITH REFETOFF SYNDROME USING METFORMIN AND THUS ACHIEVING A HEALTHY BIRTH

Abstract Introduction Refetoff syndrome (RS) is an autosomal dominantly inherited rare disease (1/40,000), characterized by thyroid hormone resistance (THR) mostly caused by Thyroid Hormone Receptor Beta (THR-B) gene defect. In the pregnancy period of female RS patients, the probability of live birth may decrease to 15%. It has been reported in the literature that metformin has effects on the thyroid hormone metabolism. Two female patients with RS being followed up in our center had pregnancy period that resulted in healthy deliveries with the use of metformin. This observation led us to question that metformin treatment may have positive effects in pregnancy of these patients by unknown mechanism of THR. We could not find any case reports or studies in the literature about the pregnancy periods in patients with RS, so we wanted to share our follow-up experience. Clinical Case Patient A and B were siblings with genetically proven THR-B mutation (figure 1). Patient A was on metformin treatment for insulin resistance before pregnancy. Her previous two pregnancies resulted in miscarriage following metformin discontinuation in pregnancy. In her third pregnancy under our follow-up, metformin treatment was continued. She completed the pregnancy without any problem and gave birth to healthy twins. Patient B, sister of Patient A, had two pregnancies. She used metformin in both pregnancies because of the favorable outcome in her sister. There were no problems in her pregnancies. The use of metformin has been shown to decrease TSH levels in patients with prediabetes and diabetes. The studies emphasize that metformin leads to a decrease in TSH values in individuals with subclinical or overt hypothyroidism, but this effect is not seen in healthy euthyroid patients. There are also reports that the effect of metformin leads to a decrease in thyroid hormone levels as well as thyroid gland and nodule size. In the literature search, we could only find a case report about metformin use in RS. In the report, metformin was started in a 28-year-old male RS patient due to a diabetic picture that developed over time. THR improved after metformin, but after a while metformin was discontinued due to dyspeptic complaints. Following the cessation of metformin THR became severe again. Although the exact mechanism of the thyroid suppressive effects of metformin is not clearly known, it is hypothesized that metformin, a compound that can cross the blood-brain barrier, increases the number and affinity of thyroid hormone receptors in the pituitary and hypothalamus, causes an increase in central dopaminergic activity, and as a result increases the pituitary feedback effect of peripheral thyroid hormones. Conclusion The incidence of healthy births in patients with RS is low. When we combine the limited literature data with our experience, we suggest that metformin treatment may contribute positively to a healthier pregnancy outcome in patients with RS.Figure 1:The pedigree of the family with Refetoff syndrome Table 1:Summary of pregnancies Table 2:Blood and genetic results of patients and* The records of two patients with Refetoff syndrome who were followed up in our center were obtained from the electronic health records.

The language of glioblastoma: a tale of cytokines and sex hormones communication

Abstract Glioblastoma (GB) is the most aggressive and frequent tumor in the central nervous system and, in humans, represents the worst prognosis for cancer. GB develops a very complex microenvironment, recruiting and interacting with a variety of cells and soluble factors, including immune cells, cytokines, and sex hormones, that contribute to GB survival and progression. Recent evidence has shown a crosstalk between cytokine and sex hormone signaling in GB. This communication could provide GB resistance to treatments and malignancy. Then, how GB orchestrates this communication is a matter of interest. For instance, a critical interaction between tumor necrosis factor beta (TGF-β) and estrogen receptor signaling has been reported in regulating epithelial-mesenchymal transition, an essential step in GB progression. Furthermore, an inhibition of TGF-β signaling by androgen receptor has been reported to promote GB tumorigenesis in men. On the other hand, it has been described that cytokines regulate steroid hormone production in different organs, and this mechanism could be involved in GB development and progression. All these data suggest an intercommunication between the immune and endocrine systems in the tumor microenvironment. Thus, in this review, we focus on explaining the knowledge about this critical intercommunication system and its implication in GB progression.

Characterization of TCRβ and IGH Repertoires in the Spleen of Two Chicken Lines with Differential ALV-J Susceptibility Under Normal and Infection Conditions

This study investigates the immunological factors underlying the differential susceptibility of two chicken strains, E- and M-lines, to avian leukosis virus subgroup J (ALV-J). During the eradication of avian leukosis at a chicken breeder farm in Guangdong, we observed strain-specific differences in susceptibility to ALV-J. Moreover, E-line chickens exhibited a slower antibody response to ALV-J compared to M-line chickens. As the T cell receptor (TCR) and B cell receptor (BCR) are critical for antigen recognition, their activation triggers specific immune responses, including antibody production. Using high-throughput sequencing, we characterized the T cell receptor beta (TCRβ) and immunoglobulin heavy chain (IGH) repertoires in spleen tissues from both chicken strains. The M-line demonstrated higher clonal diversity in both TCRβ and IGH repertoires under normal conditions compared to the E-line, suggesting a broader baseline antigen recognition capacity. Following ALV-J infection, the TCRβ repertoire diversity remained unchanged, while the IGH repertoire displayed distinct clonal expansion patterns and complementarity-determining region 3 (CDR3) length distributions between the two lines, potentially affecting their ability to recognize ALV-J antigens. Our study provides the first comprehensive comparison of TCRβ and IGH repertoire dynamics in chickens with different ALV-J susceptibilities, offering new insights into the molecular and immunological mechanisms underlying resistance to ALV-J.

Aberrant DNA methylation of EDNRB, MGMT and TIMP3 gene promoters in saliva of head and neck carcinoma patients as a diagnostic tool.

Differential DNA methylation in the promoter region of tumour suppressor genes leads to gene function silencing. In this study, we aimed to evaluate the salivary promoter methylation of EDNRB, MGMT and TIMP3 genes in H&NC patients (n = 100), premalignant lesions patients (n = 25) and healthy controls (n = 50). Blood and saliva samples were collected from all three groups and 20 concomitant tumour tissues were collected from the H&NC patients. Probe-based Methylation-specific PCR (MSP) was performed to assess the relative quantification of methylation. Significant promoter hypermethylation was detected in all three genes between H&NC patients vs. healthy controls and premalignant lesion patients vs. healthy controls. Spearman correlation analysis showed, no significant association between methylation levels and clinicopathological characteristics of HNC patients while tobacco smoking was significantly related to EDNRB methylation in premalignant lesions. The receiver operating curve (ROC) generated for EDNRB, MGMT and TIMP3 genes from saliva samples was able to differentiate between cancer vs. healthy controls and premalignant vs. healthy controls. The combined diagnostic efficiency of the panel was higher than the genes singly. The combined sensitivity of EDNRB and TIMP3 increased to 92%. This indicates that EDNRB and TIMP3 have potential value in clinical practice as effective diagnostic markers for H&NC using saliva samples.

OP11 Analysing the T cell receptor beta chain repertoire of 2,800 Inflammatory Bowel Disease patients identifies public T cell responses involved in the pathogenesis of Crohn’s Disease and Ulcerative Colitis and quantifies the impact of surgery and therapy on the immune repertoire of IBD patients

Abstract Background Crohn’s disease (CD) patients have elevated immune responses toward fungal antigens1, altered T cell responses toward gut microbiota2, and dysregulated immune responses toward Epstein-Barr virus3. Nonetheless, a detailed map of public immune responses, e.g. public T cell clonotypes associated with either CD or ulcerative colitis (UC), remains unknown. Additionally, a detailed understanding of the immune responses of different disease sub-phenotypes, e.g. ileal vs. colonic CD, and the influence of different therapies and surgical operations on the immune repertoire of IBD patients remains far from being understood. Methods We analysed the T cell receptor beta (TRB) immune repertoire of 1,890 CD patients and 914 UC patients from the US-based SPARC cohort4. Results By comparing the TRB repertoire of CD to UC patients, we identified 327 and 130 clonotypes associated with CD (Fig. 1A) and UC (Fig. 1B), respectively. Using statistical analyses, we were able to resolve the cognate HLA alleles of these clonotypes with most CD-associated clonotypes restricted to HLA-DRB1*07:01 (Fig. 1C) and UC-associated clonotypes to HLA-DRB1*15:01 alleles (Fig. 1D). CD-associated clonotypes were more expanded in ileal and ileocolonic CD relative to colonic CD and UC patients (Fig. 2A). Similarly, UC-associated clonotypes were expanded only in UC and in neither forms of CD (Fig. 2B), suggesting that different antigens and immune processes are driving CD and UC pathology. The abundance of the CD- and UC-associated clonotypes showed comparable levels in patients with and without colectomy (Fig. 2C-2D). The expansion of CD-associated clonotypes was higher in CD patients treated with Infliximab (Fig. 2E), while the expansion of UC-associated clonotypes was comparable in UC patients treated with or without Infliximab (Fig. 2F). Conclusion By analysing this comprehensive TRB dataset of IBD patients, we identified public clonotypes associated with CD and UC and resolved their cognate HLA alleles. Additionally, we identified clonotype sets associated with different disease sub-phenotypes and quantified the impact of different therapeutic procedures on their expansion. These clonotypes represent promising candidates for novel immunotherapies to treat IBD and understand its aetiology.

A Scalable, Chromatography-Free Synthesis of the Potent and Highly Selective ERβ Agonist EGX358

Abstract4-[trans-4-(Hydroxymethyl)cyclohexyl]phenol (EGX358) is a potent and highly selective estrogen receptor beta (ERβ) agonist which has demonstrated efficacy for moderation of a chemically induced hot flash and for memory consolidation in an ovariectomized mouse model for menopause. An improved synthetic route to EGX358 is reported which proceeds in four steps and which is chromatography-free.