Estrogen Receptor 1 Research Articles

A NETWORK PHARMACOLOGICAL INVESTIGATION INTO THE TREATMENT OF PERIMENOPAUSAL DEPRESSION WITH A WARMING AND TONIFYING KIDNEY-YANG RECIPE

We examine the mechanism of warming and tonifying kidney-yang recipe (WTKYR) in treating perimenopausal depression (PMD) using a network pharmacological approach. We identified the active ingredients and targets of WTKYR using the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database. PMD targets were extracted from the Online Mendelian Inheritance in Man (OMIM), DisGeNET, and GeneCards databases. Venn diagrams were created with Venny to identify common WTKYR and PMD targets. A network diagram of WTKYR targets in PMD was created with Cytoscape to examine its core active ingredients. A protein–protein interaction (PPI) network of WTKYR’s targets was constructed using the STRING database, and its primary targets were evaluated using Network Analyzer and MCODE. Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses of essential WTKYR target genes were performed with Bioconductor. The LibDock program was applied to molecularly dock the core active ingredients of the WTKYR and the critical targets of PMD. The core WTKYR components for PMD identified were as oleic acid, beta-sitosterol, and phytodolor. The core WTKYR targets identified were the Jun proto-oncogene AP-1 transcription factor subunit (JUN), peroxisome proliferator-activated receptor gamma (PPARG), brain-derived neurotrophic factor (BDNF), caspase 3 (CASP3), and estrogen receptor 1 (ESR1) proteins and the neurotrophin, estrogen, serotonergic synapse, and cyclic adenosine monophosphate (cAMP) signaling pathways. The primary functions of these pathways include extensive neuroprotective and regenerative effects, regulation of 5-hydroxytryptamine (5-HT) metabolism, and participation in regulating the physiological functions of endogenous substances. These improve synaptic sensitivity, promote neuronal cell excitability, and enhance persistent firing in the medial and lateral entorhinal cortex. Molecular docking results show good affinity between the core active ingredients of WTKYR and the critical targets of PMD. This study predicted the essential WTKYR components and its primary targets and pathways for PMD treatment, and found that improving synaptic plasticity, participating in neuroelectrophysiological processes, and regulating related signaling pathways enhance PMD, which require confirmation by future cell or animal validation experiments.

Abstract P2-03-22: Assessment of the relationship between duration of treatment with hormonal therapy and the detection of ESR1 mutations in hormone-receptor positive, HER2 negative, breast cancer

Abstract Background Hormonal therapeutic agents such as the selective estrogen receptor modulator, tamoxifen, as well as the aromatase inhibitors (AIs), anastrozole, exemestane, and letrozole, are a mainstay in the treatment of patients with hormone-receptor positive (HR+) breast cancer. Despite the effectiveness of hormonal therapy in the treatment of patients with HR+ breast cancer, acquired resistance to both AIs and tamoxifen does occur, often associated with mutations of the estrogen receptor 1 (ESR1). Although the acquisition of ESR1 mutations in patients with HR+ breast cancer treated with hormonal therapy has been established as a mechanism of resistance, there is a paucity of data on the relationship between the duration of treatment with hormonal therapies and the development of ESR1 mutations. Using a cohort of patients with HR+ breast cancer who underwent next generation sequencing (NGS) as part of routine clinical care, we aim to assess the relationship between the duration of exposure to aromatase inhibitors and the detection of ESR1 mutations. The results of this study will further our understanding of ESR1-mutations and provide insight into strategies to optimize therapy for patients with HR+ breast cancers. Methods This is a retrospective analysis of patients 18 years of age or older treated at a single academic cancer center in the USA, diagnosed with HR+, HER2-negative breast cancer and who have received somatic NGS as part of clinical care over a six-year period through July 2019. Demographic, disease-related, and treatment related variables were collected along with genomic testing variables (ie. ESR1-mutation status) for each patient who met the inclusion criteria. Two patient groups were formed on the basis of ESR1-mutation status (present vs. absent) and compared on the basis of duration of treatment with AI, and other clinical and demographic categories. Results A total of 247 patients who were diagnosed with HR-positive, HER2-negative breast cancer and received NGS were identified and are included in this analysis. A total of 138 (56%) patients were assessed via tissue based NGS and 109 (44%) by liquid biopsy. The median age at time of sequencing was 58 years [57 years (ESR1-absent) vs. 60 years (ESR1-present); p =0.009]. The majority of patients were white (83%) and post-menopausal (57%). Invasive ductal carcinoma was the most common histological type (82%). Nearly all (96%) patients had stage IV disease at time of sequencing and ESR1 mutations were found exclusively in those with stage IV disease. ESR1 mutations were identified in 62 patients (25%). The median duration of treatment with aromatase inhibitors prior to testing was 19.3 months in both groups (ESR1 mutations absent vs. detected). There was no difference in prior tamoxifen use between groups (absent 43%, present 48%; p=0.48). In patients tested by tissue based NGS, ESR1 mutations were more frequently detected in metastatic sites (28% vs. 13%; p=0.03). Twenty-two patients for whom NGS did not identify an ESR1 mutation prior to July 2019 received subsequent NGS testing. Of these 22 patients, six (27%) had ESR1 mutations detected by subsequent NGS. In these six patients, ESR1-mutations were detected by NGS a median of 2.2 years (range 1.7 years to 2.6 years) after prior NGS. Conclusion There was no difference in duration of treatment with aromatase inhibitor therapy in patients with ESR1 mutations vs. those without ESR1 mutations detected by NGS. Patients with ESR1 mutations tended to be older. Although the detection of ESR1 mutations is associated with acquired therapy resistance and progression on treatment, a longer duration of treatment may make the emergence of ESR1 mutations more likely. These counterbalancing factors may have contributed to the results reported here. Citation Format: Hannah Ayettey-Anie, Kamran A. Ahmed, Iman Washington, Kosj Yamoah, Todd Knepper. Assessment of the relationship between duration of treatment with hormonal therapy and the detection of ESR1 mutations in hormone-receptor positive, HER2 negative, breast cancer [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-03-22.

Abstract P4-09-12: Baseline and End-of-Treatment Biomarkers in Patients With PIK3CA-Mutated, Hormone Receptor-Positive, Human Epidermal Growth Factor Receptor 2-Negative Advanced Breast Cancer From BYLieve Study Cohorts A and B

Abstract Introduction: Phosphatidylinositol-4,5-bisphosphate 3-kinase catalytic subunit alpha (PIK3CA) is mutated in ~40% of patients (pts) with hormone receptor-positive (HR+), human epidermal growth factor receptor 2-negative (HER2−) advanced breast cancer (ABC). PIK3CA mutations are associated with resistance to endocrine therapy (ET) and worse overall survival. Alpelisib (ALP), an α-selective PI3K inhibitor and degrader, is indicated in combination with fulvestrant (FUL) for pts with PIK3CA-mutated (mut) HR+, HER2− ABC following progression on/after ET-based treatments. In the Phase 2, open-label, 3-cohort, noncomparative BYLieve study, clinical benefit of ALP in combination with ET was observed in the post-cyclin-dependent kinase 4/6 inhibitor (CDK4/6i) setting in pts with PIK3CA-mut, HR+, HER2− ABC. Here we report the results of a biomarker analysis using paired baseline (Cycle 1 Day 1) and end-of-treatment (EOT) circulating tumor DNA (ctDNA) samples from pts in BYLieve Cohorts A and B. Methods: In the BYLieve study, pts with PIK3CA-mut, HR+, HER2− ABC had CDK4/6i + aromatase inhibitor (Cohort A; N=127) or CDK4/6i + FUL (Cohort B; N=126) as treatment immediately prior to receiving ALP + FUL and ALP + letrozole, respectively. In this biomarker analysis, gene alterations were detected in ctDNA at baseline and EOT using next-generation sequencing (PanCancer V2 panel). Pts included in this interim analysis had confirmed PIK3CA mutations and matched baseline/EOT samples with enough sequencing coverage and ctDNA fraction to detect mutations at both time points. ctDNA fractions, tumor mutation burden (TMB) distributions, genomic landscapes, gain/loss of PIK3CA and estrogen receptor 1 (ESR1), chromosome 8/11 amplification profiles, and alterations in PI3K pathway and potential CDK4/6i resistance markers were assessed across time points. Sample sizes were small; results should thus be interpreted with caution. Results: Forty-three pts were included in the Cohort A biomarker population and 40 pts were included in Cohort B. ctDNA fraction was numerically higher at EOT compared with baseline in both cohorts; further analyses will be presented. In Cohort A, no significant differences were observed in TMB at EOT compared with baseline (P=0.21). In Cohort B, TMB was higher at EOT compared with baseline (P=0.053). Chromosome 8/11 amplifications were consistent between baseline and EOT for both cohorts. Small variations were observed in ESR1/PIK3CA mutations between baseline and EOT on both cohorts (Table). The status of potential CDK4/6i resistance markers was relatively unchanged at EOT (Table). Loss-of-function mutations in PTEN, a known PI3K inhibitor resistance marker, increased from 9% at baseline to 14% at EOT in Cohort A and from 12% at baseline to 22% at EOT in Cohort B. Conclusions: Between baseline and EOT, only small variations in gene alterations in PIK3CA-mutated HR+, HER2– ABC were observed in the post-CDK4/6i setting. As the disease progressed, increases in loss-of-function mutations in PTEN at EOT in both Cohorts A and B suggested loss of PTEN in PI3K pathway may drive resistance to ALP. Early intervention with ALP, when the tumor is particularly driven by PIK3CA oncogenic mutations and before it develops more genomic complexity, may potentially provide better clinical outcomes. Table. Gene Alteration Gain/Loss at Baseline/EOT Across Cohorts A and B Citation Format: Dejan Juric, Nicholas Turner, Sherene Loi, Fabrice Andre, Stephen K. Chia, Komal Jhaveri, Patrick Neven, Rebecca Dent, Eva Ciruelos, Mukta Joshi, Estelle Roux, Heather Patino, Murat Akdere, Hope Rugo. Baseline and End-of-Treatment Biomarkers in Patients With PIK3CA-Mutated, Hormone Receptor-Positive, Human Epidermal Growth Factor Receptor 2-Negative Advanced Breast Cancer From BYLieve Study Cohorts A and B [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P4-09-12.

Abstract P2-23-17: Programmed death ligand-1 expression in triple-negative breast cancer from Nigeria

Abstract Introduction: The incidence of triple negative breast cancer (TNBC) in West Africa appears to mirror the higher incidence of the disease among African American women in the United States. However, there remains a paucity of molecular data on TNBC from sub-Saharan Africa, despite the emergence of effective immunotherapies. Methods: Consecutive patients diagnosed with invasive breast cancer between March 2018-Jan 2020 were identified from a prospective clinical database and paired biobank at Obafemi Awolowo University Teaching Hospital (OAUTH). All specimens were processed and fixed in formalin within 60 minutes of excision by a trained pathologist. Tissue sections (4μm thick) representative of tumor were selected for routine evaluation of estrogen, progesterone and human epidermal growth factor receptor-2 expression by immunohistochemistry. This was performed at OAUTH with adequate controls according to the ASCO/CAP guidelines and verified at an outside institution for quality assurance. Additional sections from the FFPE blocks of TNBC specimens were further stained using the Dako PharmDx 22C3 PD-L1 commercial assay according to manufacturer protocols at Dalhousie University. External on-slide controls included tonsil, PD-L1 negative TNBC, and PD-L1 positive TNBC. PD-L1 expression was scored using the combined positive score (CPS), which is the number of 22C3 staining tumour cells, lymphocytes, and macrophages divided by the number of viable tumour cells, multiplied by 100. The threshold for a positive result was a CPS of ≥10 as per manufacturer instructions and institutional protocol. Research ethics board approval as well as data and material transfer agreements between institutions was obtained for this study. Results: From 85 cases, 32 were TNBC (37.6%). The mean age and BMI were 49.6±SD 7.7 and 26.2±5.7, respectively. The majority of patients presented with locally advanced disease (64.2% Stage III, 14.2% Stage IV). Seventy-nine percent (78.5%) of patients received an average of five cycles of neoadjuvant chemotherapy (SD 2.3). From 32 TNBC specimens, 27 had available FFPE tissue blocks, and of those, 24 had interpretable PD-L1 IHC for inclusion in the analysis. A total of 37.5% (9/24) of cases demonstrated a CPS ≥10. Conclusions: Over a third of breast cancer specimens in this Nigerian cohort were triple negative, 37.5% of which had PD-L1 CPS scores of ≥10. These results suggest a large proportion of patients in Nigeria may benefit from access to immunotherapy. This is the first reported incidence of PD-L1 expression in breast cancer from sub-Saharan Africa. Citation Format: Olalekan Olasehinde, Funmilola Wuraola, Aleksandra Kajetanowicz, Gilllian Bethune, Marcia Edelweiss, Peter Ntiamoah, Oluwole Odujoko, Avinash Sharma, Victoria Mango, Peter Kingham, Olusegun Alatise, Gregory Knapp. Programmed death ligand-1 expression in triple-negative breast cancer from Nigeria [abstract]. In: Proceedings of the 2022 San Antonio Breast Cancer Symposium; 2022 Dec 6-10; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2023;83(5 Suppl):Abstract nr P2-23-17.

GDNF-RET signaling and EGR1 form a positive feedback loop that promotes tamoxifen resistance via cyclin D1

BackgroundRearranged during transfection (RET) tyrosine kinase signaling has been previously implicated in endocrine resistant breast cancer, however the mechanism by which this signaling cascade promotes resistance is currently not well described. We recently reported that glial cell-derived neurotrophic factor (GDNF)-RET signaling appears to promote a positive feedback loop with the transcription factor early growth response 1 (EGR1). Here we investigate the mechanism behind this feedback loop and test the hypothesis that GDNF-RET signaling forms a regulatory loop with EGR1 to upregulate cyclin D1 (CCND1) transcription, leading to cell cycle progression and tamoxifen resistance.MethodsTo gain a better understanding of the GDNF-RET-EGR1 resistance mechanism, we studied the GDNF-EGR1 positive feedback loop and the role of GDNF and EGR1 in endocrine resistance by modulating their transcription levels using CRISPR-dCAS9 in tamoxifen sensitive (TamS) and tamoxifen resistant (TamR) MCF-7 cells. Additionally, we performed kinetic studies using recombinant GDNF (rGDNF) treatment of TamS cells. Finally, we performed cell proliferation assays using rGDNF, tamoxifen (TAM), and Palbociclib treatments in TamS cells. Statistical significance for qPCR and chromatin immunoprecipitation (ChIP)-qPCR experiments were determined using a student’s paired t-test and statistical significance for the cell viability assay was a one-way ANOVA.ResultsGDNF-RET signaling formed a positive feedback loop with EGR1 and also downregulated estrogen receptor 1 (ESR1) transcription. Upregulation of GDNF and EGR1 promoted tamoxifen resistance in TamS cells and downregulation of GDNF promoted tamoxifen sensitivity in TamR cells. Additionally, we show that rGDNF treatment activated GDNF-RET signaling in TamS cells, leading to recruitment of phospho-ELK-1 to the EGR1 promoter, upregulation of EGR1 mRNA and protein, binding of EGR1 to the GDNF and CCND1 promoters, increased GDNF protein expression, and subsequent upregulation of CCND1 mRNA levels. We also show that inhibition of cyclin D1 with Palbociclib, in the presence of rGDNF, decreases cell proliferation and resensitizes cells to TAM.ConclusionOutcomes from these studies support the hypotheses that GDNF-RET signaling forms a positive feedback loop with the transcription factor EGR1, and that GDNF-RET-EGR1 signaling promotes endocrine resistance via signaling to cyclin D1. Inhibition of components of this signaling pathway could lead to therapeutic insights into the treatment of endocrine resistant breast cancer.

Cell State Transitions and Phenotypic Heterogeneity in Luminal Breast Cancer Implicating MicroRNAs as Potential Regulators.

Luminal breast cancer subtypes respond poorly to endocrine and trastuzumab treatments due to cellular heterogeneity arising from the phenotype transitions, accounted for mainly by the loss of receptor expression. The origins of basal-like and human epidermal growth factor receptor 2 (HER2)-overexpressing breast cancer subtypes have been attributed to genetic and protein modifications in stem-like cells and luminal progenitor cell populations, respectively. The post-transcriptional regulation of protein expression is known to be influenced by microRNAs (miRNAs) that are deemed to be master regulators of several biological processes in breast tumorigenesis and progression. Our objective was to identify the fractions of luminal breast cancer cells that share stemness potentials and marker profiles and to elucidate the molecular regulatory mechanism that drives transitions between fractions, leading to receptor discordances. Established breast cancer cell lines of all prominent subtypes were screened for the expression of putative cancer stem cell (CSC) markers and drug transporter proteins using a side population (SP) assay. Flow-cytometry-sorted fractions of luminal cancer cells implanted in immunocompromised mice generated a pre-clinical estrogen receptor alpha (ERα+) animal model with multiple tumorigenic fractions displaying differential expression of drug transporters and hormone receptors. Despite an abundance of estrogen receptor 1 (ESR1) gene transcripts, few fractions transitioned to the triple-negative breast cancer (TNBC) phenotype with a visible loss of ER protein expression and a distinct microRNA expression profile that is reportedly enriched in breast CSCs. The translation of this study has the potential to provide novel therapeutic miRNA-based targets to counter the dreaded subtype transitions and the failure of antihormonal therapies in the luminal breast cancer subtype.

Material basis and mechanism of Curcuma longa tuberous roots with and without vinegar processing in treating primary dysmenorrhea

Liquid chromatography-mass spectrometry was employed to analyze the chemical components in Curcuma longa tuberous roots(HSYJ), C. longa tuberous roots processed with vinegar(CHSYJ), and rat serum after the administration. The active components of HSYJ and CHSYJ absorbed in serum were identified based on the secondary spectrum of database and literature. The targets of primary dysmenorrhea was screened out from database. The protein-protein interaction network analysis, gene ontology(GO) functional annotation, and Kyoto Encyclopedia of Genes and Genomes(KEGG) pathway enrichment analysis were performed for the common targets shared by the drug active components in serum and primary dysmenorrhea, and the component-target-pathway network was constructed. AutoDock was used to conduct molecular docking between the core components and targets. A total of 44 chemical components were identified from HSYJ and CHSYJ, including 18 absorbed in serum. On the basis of network pharmacology, we identified 8 core components(including procurcumenol, isobutyl p-hydroxybenzoate, ferulic acid, and zedoarondiol) and 10 core targets \[including interleukin-6(IL-6), estrogen receptor 1(ESR1), and prostaglandin-endoperoxide synthase 2(PTGS2)\]. The core targets were mainly distributed in the heart, liver, uterus, and smooth muscle. The molecular docking results showed that the core components were well bound to the core targets, indicating that HSYJ and CHSYJ may exert therapeutic effect on primary dysmenorrhea via estrogen, ovarian steroidogenesis, tumor necrosis factor(TNF), hypoxia-inducible factor-1(HIF-1), IL-17 and other signaling pathways. This study clarifies the HSYJ and CHSYJ components absorbed in serum, as well as the corresponding mechanism, providing a reference for further elucidating the therapeutic material basis and clinical application of HSYJ and CHSYJ.

COMPARISON OF ANDROGEN AND ESTROGEN RECEPTORS’ EXPRESSION IN DARTOS TISSUE OF BOYS WITH AND WITHOUT HYPOSPADIAS

Objective: This study aims to investigate the characteristics of androgen receptors (AR), estrogen receptor 1 (ER1) and estrogen receptor 2 (ER2) expression in dartos tissue of patients with congenital hypospadias, compared to normal penis. Material & Methods: We harvested 63 dartos tissue consisting of 53 congenital hypospadias that underwent urethroplasty (20 distal and 33 proximal) and 10 normal penis that underwent circumcision as controls from September 2017 to September 2018. The expressions of AR, ER1, and ER2 were measured using Quantitative Real-Time PCR (qPCR). All data were analyzed by Prism 7, and one-way ANOVA tests were used to compare gene expressions between the groups. Results: The mean age was 68.99 (± 45.5) and 65.6 (± 25.8) months in boys with and without hypospadias, respectively. The expression of mRNA AR was decreased in proximal (6.26 ± 2.30) and distal hypospadias (6.43 ± 2.22) compared to controls (9.69 ± 1.10), which were statistically significant (p=0.0001 and p<0.0001, respectively). We found a statistically significant difference of ER1 expression compared to controls (p=0.0064). The expression of ER2 was significantly increased in distal (21.03 ± 5.00) and proximal hypospadias (25.21 ± 8.06) groups compared to controls (11.80 ± 2.49) (p<0.0001). There was no statistically significant mean difference in mRNA ER1 expression (p=0.65). Conclusion: The repressed AR and elevated ER mRNA as shown in our study may suggest that defects in those receptors’ interaction and/or balance may contribute to hypospadias and penile curvature condition. Further studies are needed to evaluate any gene-related problems in hypospadias.
 Keywords: Hypospadias, androgen receptor, estrogen receptor, dartos tissue.

Transcriptional regulatory network during axonal regeneration of dorsal root ganglion neurons: laser-capture microdissection and deep sequencing.

The key regulators and regeneration-associated genes involved in axonal regeneration of neurons after injury have not been clarified. In high-throughput sequencing, various factors influence the final sequencing results, including the number and size of cells, the depth of sequencing, and the method of cell separation. There is still a lack of research on the detailed molecular expression profile during the regeneration of dorsal root ganglion neuron axon. In this study, we performed laser-capture microdissection coupled with RNA sequencing on dorsal root ganglion neurons at 0, 3, 6, and 12 hours and 1, 3, and 7 days after sciatic nerve crush in rats. We identified three stages after dorsal root ganglion injury: early (3-12 hours), pre-regeneration (1 day), and regeneration (3-7 days). Gene expression patterns and related function enrichment results showed that one module of genes was highly related to axonal regeneration. We verified the up-regulation of activating transcription factor 3 (Atf3), Kruppel like factor 6 (Klf6), AT-rich interaction domain 5A (Arid5a), CAMP responsive element modulator (Crem), and FOS like 1, AP-1 transcription factor Subunit (Fosl1) in dorsal root ganglion neurons after injury. Suppressing these transcription factors (Crem, Arid5a, Fosl1 and Klf6) reduced axonal regrowth in vitro. As the hub transcription factor, Atf3 showed higher expression and activity at the pre-regeneration and regeneration stages. G protein-coupled estrogen receptor 1 (Gper1), interleukin 12a (Il12a), estrogen receptor 1 (ESR1), and interleukin 6 (IL6) may be upstream factors that trigger the activation of Atf3 during the repair of axon injury in the early stage. Our study presents the detailed molecular expression profile during axonal regeneration of dorsal root ganglion neurons after peripheral nerve injury. These findings may provide reference for the clinical screening of molecular targets for the treatment of peripheral nerve injury.

Bioinformatics-based analysis of the roles of sex hormone receptors in endometriosis development.

Endometriosis is a hormone-dependent disease in women of reproductive age and seriously affects women's health. To analyze the involvement of sex hormone receptors in endometriosis development, we performed bioinformatics analysis using four datasets derived from the Gene Expression Omnibus (GEO) database, which may help us understand the mechanisms by which the sex hormones act in vivo in endometriosis patients. The enrichment analysis and protein-protein interaction (PPI) analysis of the differentially expressed genes (DEGs) revealed that there are different key genes and pathways involved in eutopic endometrium aberrations of endometriosis patients and endometriotic lesions, and sex hormone receptors, including androgen receptor (AR), progesterone receptor (PGR) and estrogen receptor 1 (ESR1), may play important roles in endometriosis development. Androgen receptor (AR), as the hub gene of endometrial aberrations in endometriotic patients, showed positive expression in the main cell types for endometriosis development, and its decreased expression in the endometrium of endometriotic patients was also confirmed by immunohistochemistry (IHC). The nomogram model established based on it displayed good predictive value.

Association study of ESR1 rs9340799, rs2234693, and MMP2 rs243865 variants in Iranian women with premature ovarian insufficiency: A case-control study.

Primary ovarian insufficiency (POI) is a rare disease clinically characterized by ovarian follicles depletion or dysfunction and menopause before the age of 40 yr as the cut-off age for POI. It is a complex disease, and its etiology involves several factors. However, genetic factors have a predominant role in the susceptibility to the disease. This study aims to investigate the polymorphisms of rs243865 in the matrix metallopeptidase 2 (MMP2) gene and rs2234693 and rs9340799 in the estrogen receptor 1 (ESR1) gene with susceptibility to POI in Iranian women under 35 yr. This case-control study was performed on 150 women with POI and 150 healthy women who were referred to Yazd Reproductive Sciences Institute, Yazd, Iran between May-October 2020. The genotyping of ESR1 rs9340799, rs2234693, and MMP2 rs243865 polymorphism was done using tetra-amplification refractory mutation system-polymerase chain reaction. In addition, haplotype analysis and linkage disequilibrium were investigated by SNPanalyzer software. Our study revealed the frequency of rs243865 TT, CC genotypes in the MMP2 gene and rs2234693 CC, TT; and rs9340799 GG, AA in the ESR1 gene were more prevalent in the case group compared to the control group. In addition, ESR1 rs2234693 and rs9340799 genotypes showed significant association with the development of the disease in our population. Among 4 haplotypes for 2 polymorphisms in the ESR1 gene, rs2234693T/rs9340799A haplotype was associated with conferring risk to POI. ESR1 rs2234693 and rs9340799 polymorphism were strongly associated with our population's POI.

Isorhamnetin inhibits progression of ovarian cancer by targeting ESR1

Although reports suggest Chinese herbal medicine treatment of ovarian cancer (OC) has a good effect, the role of isorhamnetin (ISO), a flavonol aglycone with immune, anti-inflammatory, cardiovascular and cerebrovascular protective effects, as well as an anticancer effect, in OC remains unclear. Network pharmacology was used to explore this in vitro and in vivo, and to identify relevant targets. The common targets of ISO in the treatment of OC were screened by constructing drug targets and disease gene databases for Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. The protein-protein interaction network was constructed by STRING. Overlapping targets were further analyzed using the online tool UALCAN to analyze the correlation between gene expression and patient survival and prognosis. The effect of ISO on OC cell proliferation, migration, and invasion was assessed in vivo and in vitro, and the function of the estrogen receptor 1 (ESR1) in the development of OC was examined by overexpressing and knocking down ESR1 expression. Through network pharmacology analysis, 25 target genes related to ISO-OC were screened out. The overall survival rate of OC patients only significantly correlated with high expression of ESR1 among 13 highly expressed overlapping genes. ISO significantly inhibited the proliferation, migration and invasion of OC cells in vitro and inhibited tumor growth in vivo. Overexpression of ESR1 significantly promoted the proliferation, migration and invasion of OC cells, whereas knockdown of ESR1 showed the opposite result. In addition, overexpression of ESR1 significantly reversed the inhibitory effect of ISO on the proliferation, migration and invasion of OC cells. We confirmed that ISO inhibits OC cell proliferation, migration and invasion by targeting ESR1 expression, which provides a theoretical basis for further pharmacological research.

Involvement of the G-Protein-Coupled Estrogen Receptor-1 (GPER) Signaling Pathway in Neurodegenerative Disorders: A Review.

The G-protein-coupled estrogen receptor-1 (GPER) is an extranuclear estrogen receptor that regulates the expression of several downstream signaling pathways with a variety of biological actions including cell migration, proliferation, and apoptosis in different parts of the brain area. It is endogenously activated by estrogen, a steroidal hormone that binds to GPER receptors which help in maintaining cellular homeostasis and neuronal integrity as well as influences neurogenesis. In contrast, neurodegenerative disorders are a big problem for society, and still many people suffer from motor and cognitive impairments. Research to date reported that GPER has the potential to whittle down motor abnormalities and cognitive dysfunction by limiting the progression of neurodegenerative disorders. Although several findings suggest that GPER activation accelerated transcription of the PI3K/Akt/Gsk-3β and ERK1/2 signaling pathway that halt disease progression by decreasing oxidative stress, neuroinflammation, and apoptosis. Accordingly, the goal of this review is to highlight the basic mechanism of GPER signaling pathway-mediated neuroprotection in various neurodegenerative disorders including Parkinson's disease (PD), Huntington's disease (HD), Tardive dyskinesia (TD), and Epilepsy. This review also discusses the role of the GPER activators which might be a promising therapeutic target option to treat neurodegenerative disorders. All the data were obtained from published articles in PubMed (353), Web of Science (788), and Scopus (770) databases using the search terms: GPER, PD, HD, TD, epilepsy, and neurodegenerative disorders.

Haplotype of ESR1 and PPARD Genes Is Associated with Higher Anthropometric Changes in Han Chinese Obesity by Adjusting Dietary Factors-An 18-Month Follow-Up.

The obesity genetic effect may play a major role in obesogenic environment. A combined case–control and an 18-month follow-up were carried out, including a total of 311 controls and 118 obese cases. All participants were aged in the range of 20–55 y/o. The body mass index (BMI) of obese cases and normal controls was in the range of 27.0–34.9 and 18.5–23.9 kg/m2, respectively. The rs712221 on Estrogen receptor1 (ESR1) and rs2016520 on Peroxisome proliferator–activated receptor delta (PPARD) showed significant associations with obesity. The TT (odds ratio (OR): 2.42; 95% confidence interval (CI): 1.46–4.01) and TT/TC (OR: 2.80; 95% CI: 1.14–6.85) genotypes on rs712221 and rs2016520 had significantly higher obesity risks, respectively. Moreover, the synergic effect of these two risk SNPs (2-RGH) exhibited an almost geometrical increase in obesity risk (OR: 7.00; 95% CI: 2.23–21.99). Obese individuals with 2-RGH had apparently higher changes in BMI increase, body weight gain and dietary fiber intake but a lower total energy intake within the 18-month follow-up.

Genetic Polymorphisms of Estrogen Receptors α and β are Associated with Craniofacial Measurements in Patients With Dentofacial Deformity.

Dentofacial deformities are characterized by abnormalities in craniofacial development that affects the individual's skeletal and occlusion, often causing functional and esthetic problems. In literature, there is an involvement of polymorphisms in estrogen receptor 1 (ESR1) and estrogen receptor 2 (ESR2) genes in craniofacial measurements. The aim of this study was to evaluate a possible association between polymorphisms in ESR1 (rs2234693 and rs9340799) and ESR2 (rs1256049 and rs4986938) genes with cephalometric measurements in individuals with dentofacial deformities. This cross-sectional study was performed with 158 individuals in the preoperative period of orthognathic surgery. The cephalometric measurements obtained through lateral cephalogram using Dolphin Imaging software. For genetic analysis, the DNA extracted from epithelial cells of the oral mucosa and were genotyped using the real-time polymerase chain reaction. The data found submitted to statistical analysis, through the Kolmogorov-Smirnov, Mann-Whitney, and Kruskal-Wallis tests, using the IBM SPSS software version 24.0. Considered a significance level of 0.05. We found association between polymorphisms and cephalometric measurements just in the female sex. The polymorphisms ESR1/rs9340799 ( P= 0.003), ESR1/rs2234693 ( P= 0.026), and ESR2/rs1256049 ( P= 0.046) were associated with the upper gonial angle (Ar-Go-N). The polymorphism ESR2/rs1256049 was also associated with the facial axis-rickets (NBa-PtGn) ( P= 0.004), anterior cranial base (SN) ( P= 0.036), and Y-axis (SGn-SN) ( P= 0.031).

Screening of key biomarkers in osteoporosis: Evidence from bioinformatic analysis.

To analyze the expression characteristics of osteoporosis-related genes by bioinformatics and elucidate the pathogenesis of osteoporosis. The differentially expressed genes (DEGs), microRNA (miRNA), and genes with differentially methylated regions (DMRS) in promoters were identified. The protein-protein interaction (PPI) network was constructed and performed. The Clue Gene Ontology analysis and miRNA-mRNA (messenger RNA) regulatory network were constructed using Cytoscape. Fifty-nine DEGs, 10 differential miRNAs, and 2083 genes with DMRs were screened out. The Proteasome-Modulator (PSMD) family proteins and estrogen receptor 1 (ESR1) are vital for the PPI analysis of DEGs. The interaction network of the Smad3 protein showed that the degree of connection to ESR1, PSMD11, and transcription factor 4 (TCF4) is very high. Homo sapiens (hsa)-miR-106b-5p was differential and regulated TCF4 through building the miRNA-mRNA regulatory network. Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment of DEGs focused on vascular smooth muscle contraction, thyroid hormone signaling pathway, and estrogen signaling pathway. The Gene Ontology (GO) function analysis of genes with DMRs in promoters was primarily concentrated in the cell differentiation, positive regulation of CDP-diacylglycerol-serine O-phosphatidyltransferase activity, and positive regulation of C-palmitoyltransferase activity. The KEGG enrichment of genes with DMRs in promoters largely focused on glycerol phospholipid metabolism, histidine metabolism, Adenosine 5'-monophosphate-activated protein kinase signaling pathway, Hedgehog signaling pathway, and mRNA surveillance pathway. Hsa-miRNA-106b-5p regulates bone formation and the pathogenesis of osteoporosis by controlling TCF4, and methylation modification of TCF4 can also affect the pathogenesis of osteoporosis.

Estrogen receptor 1 inhibits the progression of hepatocellular carcinoma via positively regulating lncRNA maternally expressed gene 3 under high glucose conditions.

Long noncoding RNA (lncRNA) maternally expressed gene 3 (MEG3) is crucial in the association of diabetes mellitus (DM) and hepatocellular carcinoma (HCC), and estrogen receptor 1 (ESR1) plays an essential role in various cancers. However, the underlying regulatory effect of ESR1/lncRNA MEG3 on HCC with DM remains unclear. This study explored the regulatory effect of ESR1/lncRNA MEG3 on HCC cell progression. Bioinformatics analysis was used to predict the promoter sequence of lncRNA MEG3 using UCSC (http://genome.ucsc.edu/), followed by luciferase reporter and RNA immunoprecipitation (RIP) assays to verify the specific combination between ESR1 and lncRNA MEG3 promoter. After cotransfection with ESR1, ESR1 siRNA or lncRNA MEG3 RNA, CCK-8, 5-ethynyl-2'-deoxyuridine (EdU) and colony formation assays were used to evaluate the cell proliferation capacity. Cell apoptosis was assessed using flow cytometry analysis. Next, wound healing and Transwell assays were conducted to examine cell invasiveness and migration. Quantitative real-time polymerase chain reaction (qRT-PCR) and western blot analysis were performed to quantify the expression of ESR1 or lncRNA MEG3. ESR1 might be the transcription factor (TF) of lncRNA MEG3, and ESR1 bound with lncRNA MEG3 promoter. Overexpression of ESR1 repressed the proliferation, migration and invasion of HepG2 cells, and promoted apoptosis of HepG2 cells under high glucose conditions. Silencing ESR1 decreased lncRNA MEG3 expression in HepG2 cells but enhanced proliferation, migration and invasion. Meanwhile, a rescue assay demonstrated that silencing lncRNA MEG3 reversed the inhibitory effect of ESR1 on HepG2 cell progression. ESR1 inhibits HCC cell progression through positively regulating lncRNA MEG3, and the results provide a promising strategy for HCC management.

Estrogen enforces the integrity of blood vessels in the bone during pregnancy and menopause

The mammalian skeletal system shows sex differences in structure, functions, aging and disease incidences. The role of blood vessels in physiological, regenerative and pathological bone functions indicates the requisite to understanding their sex specificity. In this study, we found that estrogen regulates blood vessel physiology during pregnancy and menopause through estrogen receptor alpha (ERα) and G-protein-coupled estrogen receptor-1 (GPER1) but not ERβ-dependent signaling in mice. Estrogen regulates the lipid use of bone endothelial cells (BECs) and promotes lipolysis of adipocytes and fatty acid (FA) uptake from the microenvironment. Low estrogen conditions skew endothelial FA metabolism to accumulate lipid peroxides (LPOs), leading to vascular aging. High ferrous ion levels in female BECs intensify LPO accumulation and accelerate the aging process. Notably, inhibiting LPO generation using liproxstatin-1 in aged mice significantly improved bone heath. Thus, our findings demonstrate the effects of estrogen on BECs and suggest that LPO targeting could be an efficient strategy to manage blood and bone health in females.

Molecular characterization of ESR1 variants in breast cancer.

Estrogen receptor 1 (ESR1) mutations and fusions typically arise in patients with hormone receptor-positive breast cancer after aromatase inhibitor therapy, whereby ESR1 is constitutively activated in a ligand-independent manner. These variants can impact treatment response. Herein, we characterize ESR1 variants among molecularly profiled advanced breast cancers. DNA next-generation sequencing (592-gene panel) data from 9860 breast cancer samples were retrospectively reviewed. Gene fusions were detected using the ArcherDx fusion assay or whole transcriptome sequencing (n = 344 and n = 4305, respectively). Statistical analyses included Chi-square and Fisher's exact tests. An ESR1 ligand-binding domain (LBD) mutation was detected in 8.6% of tumors evaluated and a pathogenic ESR1 fusion was detected in 1.6%. Most ESR1 LBD mutations/fusions were from estrogen receptor (ER)-positive samples (20.1% and 4.9%, respectively). The most common ESR1 LBD mutations included D538G (3.3%), Y537S (2.3%), and E380Q (1.1%) mutations. Among biopsy sites, ESR1 LBD mutations were most observed in liver metastases. Pathogenic ESR1 fusions were identified in 76 samples (1.6%) with 40 unique fusion partners. Evaluating co-alterations, ESR1 variant (mutation/fusion) samples more frequently expressed androgen receptor (78.0% vs 58.6, P < 0.0001) and less frequently immune checkpoint proteins than ESR1 wild-type (PD-1 20.0% vs 53.4, P < 0.05; immune cell PD-L1 10.0% vs 30.2, P < 0.0001). We have described one of the largest series of ESR1 fusions reported. ESR1 LBD mutations were commonly identified in ER-positive disease. Limited data exists regarding the clinical impact of ESR1 fusions, which could be an area for future therapeutic exploration.

Network Pharmacology-Integrated Molecular Docking Reveals the Expected Anticancer Mechanism of Picrorhizae Rhizoma Extract.

This study sought to explore the anticancer mechanism of Picrorhizae Rhizoma (PR) extract based on network pharmacology and molecular docking. The potential chemicals of PR were screened through the Traditional Chinese Medicine Systems Pharmacology (TCMSP) database and relevant literatures. Corresponding targets of active ingredients were found with the help of the UniProtKB database, and therapeutic targets for cancer action were screened with the help of the GeneCards database. We used Cytoscape software to construct the compound-target-pathway network of PR extract. We utilized the STRING database to obtain the protein-protein interaction (PPI) network. We used DAVID database combining Gene Ontology (GO) analysis and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis. Finally, molecular docking was employed for initial efficacy checking. We have identified 16 potential active components of PR through screening, involving 112 disease action targets. Utilizing the GeneCards database, 112 intersecting targets between PR extract and cancer were found, which mainly exerts anticancer effects by regulating tumor necrosis factor (TNF), recombinant caspase 3 (CASP3), c-Jun NH2-terminal kinase (JNK)/JUN, epidermal growth factor receptor (EGFR), and estrogen receptor-1 (ESR1) with some other target genes and pathways associated with cancer. The major anticancer species are prostate cancer, colorectal cancer, small cell lung cancer, etc. In the molecular docking study, herbactin had a strong affinity for TNF. Based on network pharmacology and molecular docking studies, PR and their compounds have demonstrated potential anticancer activities against several key targets. Our preliminary findings provide a strong foundation for further experiments with PR constituents.