Estrogen Receptor Alpha Protein Research Articles

Menstrual and oral contraceptive pill cycles minimally influence vascular function and associated cellular regulation in premenopausal females.

Historical exclusion of females in research has been, in part, due to the perceived influence of natural menstrual (NAT) and oral contraceptive pill (OCP) cycles on vascular outcomes. NAT and OCP cycle phases may influence brachial artery (BA) endothelial function, however, findings are mixed. Minimal research has examined arterial stiffness, smooth muscle, and lower limb endothelial function. The purpose of this study was to investigate the influence of NAT and OCP cycles on cardiovascular outcomes and cellular regulation. Forty-nine premenopausal females (n = 17 NAT, n = 17 second generation OCP, n = 15 third generation OCP) participated in two randomized order visits in the low (LH, early follicular/placebo) and high (HH, midluteal/active) hormone cycle phases. BA and superficial femoral artery (SFA) endothelial function [flow-mediated dilation (FMD) test], smooth muscle function (nitroglycerine-mediated dilation test), and carotid and peripheral (pulse wave velocity) arterial stiffness were assessed. Cultured female human endothelial cells were exposed to participant serum for 24 h to examine endothelial nitric oxide synthase (eNOS) and estrogen receptor-α (ERα) protein content. BA FMD was elevated in the HH vs. LH phase, regardless of group (HH, 7.7 ± 3.5%; LH, 7.0 ± 3.3%; P = 0.02); however, allometric scaling for baseline diameter resulted in no phase effect (HH, 7.6 ± 2.6%; LH, 7.1 ± 2.6%; P = 0.052, d = 0.35). SFA FMD, BA, and SFA smooth muscle function, arterial stiffness, and eNOS and ERα protein content were unaffected. NAT and OCP phases examined have minimal influence on vascular outcomes and ERα-eNOS pathway, apart from a small effect on BA endothelial function partially explained by differences in baseline artery diameter. NEW & NOTEWORTHY Comprehensive evaluation of the cardiovascular system in naturally cycling and second and third generation OCP users indicates no major influence of hormonal phases examined on endothelial function and smooth muscle function in the arteries of the upper and lower limbs, arterial stiffness, or underlying cellular mechanisms. Study findings challenge the historical exclusion of female participants due to potentially confounding hormonal cycles; researchers are encouraged to consider the hormonal environment in future study design.

Gestational exposure to BPA alters the expression of glucose and lipid metabolic mediators in the placenta: Role in programming offspring for obesity

Bisphenol A (BPA) exposure during pregnancy is known to predispose offspring to obesity in later life. Our previous studies demonstrated obesogenic effects in BPA-exposed offspring, including excess body fat, increased feed efficiency, adipocyte hypertrophy, and altered leptin signaling. However, the role of the placenta in mediating these effects remained unclear. This study investigates the mechanisms by which BPA exposure affects placental glucose and lipid transporters and their impact on offspring adiposity in Wistar rats. Dams were orally gavaged with BPA [0.4 (low dose-LD) and 4.0 (high dose-HD) μg/kg body weight] from gestational day (gD) 4–14. Gestational exposure to LD BPA increased the expression of 11β hydroxysteroid dehydrogenase 1 (11β HSD1) and estrogen receptor alpha (ERα) proteins (p<0.05) in the placenta compared to control and HD BPA. Similar changes were observed in the expression of mTOR signaling mediators, fatty acid transporters, and intracellular fatty acid-binding proteins. There were no changes in the dam's body weight or lipid and glucose profiles. However, there was a dose dependent increase in glucose transporter (GLUT1) expression in the placenta. While LD BPA increased hexokinase 2 expression in the placenta, HD BPA had no effect. Both doses of BPA increased IL6 expression, but only LD BPA exposure increased PPAR-gamma expression. Additionally, BPA exposure induced ADRP expression and localization, suggesting potential lipid overload in the placenta. Furthermore, BPA exposure altered the placental epigenetic profile, with increased expression of DNA methyltransferases (DNMTs). Overall, gestational BPA exposure led to dose-specific alterations in placental glucose and lipid metabolic activities, possibly playing an role in increasing the supply of these macronutrients to the fetus and predisposing the offspring to obesity.

The effect of Mongolian Medicine Chagan Gaoyou-4 Powder on bone mineral density induced by retinoic acid in rats

Objective: To investigate the effects of Chagan-Gaoyou-4 powder on spine bone mineral density and serum estrogen and estrogen receptor alpha (ERa) and estrogen receptor beta (ERβ) protein expression in bone tissue induced by retinoic acid in rats. Methods: Sixty 3-month-old SPF female SD rats were randomly divided into model group (retinoin group), normal group (SHAM group), Mongolian medicine group (Chagan-Gaoyou 4 group), and control group (Gushukang granule group). After 14 days of administration, spinal Bone Mineral Density (BMD) was measured, and external manifestations were observed. Results: Compared with the SHAM group, the BMD values of all medication groups decreased (P &lt; 0.05, P &lt; 0.001). Compared with the model group, the spinal BMD value of the Chagan Gaoyou-4 group was increased after 21 days of treatment, and the difference was statistically significant (P &lt; 0.01, P &lt; 0.05). After 21 days, the spinal BMD value of all treatment groups was increased (P &lt; 0.05), the spinal BMD value was also increased, especially Chagan Gaoyou-4 group was more obviously increased (P &lt; 0.05). Compared with the normal group, ERa and ERβ levels in the model group were significantly decreased (P &lt; 0.001), and ERa and ERβ levels in all drug groups were significantly increased (P &lt; 0.001) compared with the model group. After 21 days of treatment, there was no significant difference among the three treatment groups (P &gt; 0.05). ERa and ERβ levels in the Chagan Gaoyou-4 group were higher than those in the model group (P &lt; 0.05), but there was no significant difference between the group and the Gushukang granule group (P &gt; 0.05). The E2 content of the Chagan Gaoyou-4 group was higher than that of the model group (P &lt; 0.05). The mRNA expression of estrogen receptor ERa and ERβ in bone tissue of the left proximal femur was detected by RT-PCR. Compared with the model group, the expression of ERa and ERβ mRNA in the Chagan Gaoyou-4 group and Gushukang granule group was increased. Compared with the model group, ERp mRNA expression was increased in the Chagan Gaoyou-4 powder medium-dose group. Compared with the Gushukang granule group, there was no significant difference in ERm RNA expression in the Chagan Gaoyou-4 dose groups. Chagan Gaoyou-4 can up-regulate the expression of ERa and ERβ induced by retinoic acid in rats, indicating that the Chagan Gaoyou-4 powder group may promote bone formation, regulate bone resorption, improve bone mineral density, and achieve the purpose of preventing and treating OP by increasing estrogen level, stimulating estrogen receptor, and increasing the expression of ERa and ERβ in bone tissue. Conclusion: Chagan-Gaoyu-4 powder may have estrogen-like effects on the bone tissue of rats induced by retinoic acid and may increase the level of serum estrogen, ERa, and ERβ protein expression, thereby improving the spinal BMD of experimental rats. The Chagan-Gaoyou-4 powder group could improve the general condition of osteoporosis induced by retinoic acid in rats.

Plasmonic sensor combined with a microcuvette device for monitoring molecule binding processes at ultra-low concentrations

A novel sensing strategy is presented to monitor receptor-target pair interactions at ultra-low concentrations without functionalization processes. The biosensing is made by a sensitive chip and a surface plasmon resonance (SPR) probe connected in series, both built up on multimode plastic optical fibers (POFs). The SPR probe is a conventional D-shaped POF platform, while the capture platform consists of three microholes, containing a few hundred nanoliters each, made in the core of a modified POF. The microholes are filled with a specific receptor solution, which selectively captures the target present in the samples dropped over the filled microholes. Any variation occurring in the microholes due to the molecule binding processes changes the mode profile of the propagated light in the POF's core, modifying the plasmonic effects proportionally to the phenomena under scrutiny. As a proof of concept, the interactions of two receptor-target pairs, estradiol and cortisol to the respective receptors, namely Estrogen Receptor alpha protein (ER) and Glucocorticoid Receptor (GR), were monitored over time in the attomolar range. The receptor-target pair interactions are observed at ultra-low concentrations (order of aM) by monitoring the resonance wavelength shift over time. The proposed sensing approach stands for a novel class of laboratory instrumentation with unprecedented capabilities in terms of compactness, ultra-high sensitivity and low-cost.

Sexual Dimorphism in Mitochondrial Metabolism and Insulin Sensitivity Is Controlled by HSP72-ERa Crosstalk

HSP72 is highly induced during exercise, and muscle expression is reduced in obese and diabetic men. Muscle-specific HSP72 overexpression protects male mice against diet-induced obesity and insulin resistance. The current study objective was to determine the mechanisms by which HSP72 controls metabolic function and to ascertain whether these mechanisms are impacted by biological sex. We interrogated the impact of biological sex and estradiol action in the context of HSP72 loss and gain of expression in mice. HSP72 is a binding partner of the mitochondrial quality control protein Parkin. HSP72-KO led to metabolic dysfunction via impaired Parkin action, by promoting the retention of enlarged, fission incompetent mitochondria with reduced respiratory capacity. In contrast, female KOs remained lean and insulin sensitive with small spherical muscle mitochondria. Muscle estrogen receptor alpha transcript and protein (Esr1/ERa) was induced in female HSP72-KO vs. wildtype (WT), and this enhanced Parkin-DRP1-induced fatty acid oxidation and underpinned improvements in exercise capacity. The phenotype of female HSP72-KO mice recapitulates that of muscle-specific ESR1 overexpression animals. Because muscle ERa expression was identical between male genotypes, KO vs. WT, we treated male KO mice with 17β-estradiol and the treatment restored mitochondrial form/function as well as insulin sensitivity. In summary, biological sex is critical for understanding the impact of HSP72 in governing metabolic health and insulin sensitivity. National Institutes of Health DK060484 and P30DK063491. This is the full abstract presented at the American Physiology Summit 2024 meeting and is only available in HTML format. There are no additional versions or additional content available for this abstract. Physiology was not involved in the peer review process.

MRNA PROTACs: engineering PROTACs for high-efficiency targeted protein degradation.

Proteolysis-targeting chimeras (PROTACs) are essential bifunctional molecules that target proteins of interest (POIs) for degradation by cellular ubiquitination machinery. Despite significant progress made in understanding PROTACs' functions, their therapeutic potential remains largely untapped. As a result of the success of highly flexible, scalable, and low-cost mRNA therapies, as well as the advantages of the first generation of peptide PROTACs (p-PROTACs), we present for the first time an engineering mRNA PROTACs (m-PROTACs) strategy. This design combines von Hippel-Lindau (VHL) recruiting peptide encoding mRNA and POI-binding peptide encoding mRNA to form m-PROTAC and promote cellular POI degradation. We then performed proof-of-concept experiments using two m-PROTACs targeting two cancer-related proteins, estrogen receptor alpha and B-cell lymphoma-extra large protein. Our results demonstrated that m-PROTACs could successfully degrade the POIs in different cell lines and more effectively inhibit cell proliferation than the traditional p-PROTACs. Moreover, the in vivo experiment demonstrated that m-PROTAC led to significant tumor regression in the 4T1 mouse xenograft model. This finding highlights the enormous potential of m-PROTAC as a promising approach for targeted protein degradation therapy.

Opportunities and prospects for the treatment of hormone-dependent breast cancer

The high incidence of breast cancer requires increased attention to the problem of rational pharmacotherapy of this condition. When choosing tactics for the treatment of breast cancer, it is recommended to take into account the immunohistochemical subtype of cancer cells. In the case of detection of estrogen-positive expression, an obligatory component of pharmacotherapy is endocrine therapy with antiestrogen orientation. Despite the fact that there are already many years of positive experience with the use of selective estrogen receptor modulators and aromatase inhibitors, the search for new more effective agents continues in terms of prolonging the life of patients and reducing the risk of adverse reactions.&#x0D; In recent years, cyclin-dependent kinase 4/6 inhibitors have been added to first-line therapy, which is a breakthrough in the treatment of metastatic breast cancer. The introduction of combined antiestrogen therapy with targeted agents that have the ability to inhibit phosphatidylinositol-3-kinase is logical in the event of resistance to primary endocrine therapy. However, the need to search and study new drugs remains. In this regard, the most promising direction is the development of agents that can reduce the expression of the Estrogen receptor alpha protein, and block estrogen-dependent and independent estrogen receptor signaling.

Revealing Propolis Potential Activity on Inhibiting Estrogen Receptor and Heat Shock Protein 90 Overexpressed in Breast Cancer by Bioinformatics Approaches.

Breast cancer is the most commonly diagnosed cancer globally, with the highest incidence of breast cancer occurring in Asian countries including Indonesia. Among the types of breast cancer, the estrogen receptor (ER)-positive subtype which is prominent with estrogen receptor alpha (ERα) and heat shock protein 90 (HSP90) overexpression genes becomes the most prevalent than the others, approximately 75% of all breast cancer cases. ERα and HSP90 play a role in breast cancer activities including breast tumor growth, invasion, and metastasis mechanism. Propolis, a natural bee product, has been explored for its anticancer activity. However, there is lack of studies that evaluated the potential inhibitor from propolis compounds to the ERα and HSP90 proteins. Therefore, this article focuses on examining the correlation between ERα and HSP90's role in breast cancer and investigating the potential of 93 unique propolis compositions in inhibiting these genes in breast cancer using in silico approaches. This study revealed the positive correlation between ERα and HSP90 genes in breast cancer disease development. Furthermore, we also found novel potential bioactive compounds of propolis against breast cancer through binding with ERα and HSP90; they were 3',4',7-trihydroxyisoflavone and baicalein-7-O-β-D glucopyranoside, respectively. Further research on these compounds is needed to elucidate deeper mechanisms and activity in the real biological system to develop new breast cancer drug treatments.

Phytogenic One-pot Synthesis and Spectroscopic Characterization of Novel Mono Benzylated Resveratrol Hybrid Molecule Using Extracted Resveratrol from Green Grape Peels: In Silico ADMET Study and In Vitro Antitumor Activities Against Breast Cancer Cells

Background: Despite the development of numerous customized techniques for treating breast cancer, cancer patients' clinical results revealed adverse consequences in addition to chemotherapeutic drug resistance. Hence, finding therapeutic compounds with little or no side effects becomes essential in the fight against cancer. Resveratrol, a naturally occurring non-flavonoid polyphenol present in plants as a phytoalexin, is a promising therapeutic agent that has garnered the interest of several researchers due to its prodigious pharmacological and biological activities, but its unfavourable pharmacokinetic properties complicated its clinical studies. Along with several structural modifications, substitutions, etc., that have already been reported, this is the first time that a novel resveratrol analogue comprising an aromatic hetero moiety (ResD1) was synthesized using resveratrol isolated from grape (Vitis vinifera) peels as a precursor. Methods: ResD1 was synthesized by one-pot reaction using extracted and isolated resveratrol from grape peels. Structure confirmation of the isolated resveratrol and synthesized resveratrol derivative was elucidated by 1H-NMR, 13C-NMR, FTIR, and LC-MS. In silico molecular docking and ADMET study of ResD1 were carried out using AutoDock 4.2 and ADMETLab 2.0. ResD1 was evaluated for in vitro antioxidant, antimicrobial, anticancer, and CAXII gene expression as per the standard methods. Results: In silico molecular docking results revealed that ResD1 is capable of attaching to the ERα (estrogen receptor alpha) protein via hydrogen and hydrophobic bonds and has -7.12 kcal/mol as docking score. The novel derivative (IC50 = 42.8 µg/ml) exhibited better radical scavenging ability than ascorbic acid (control). The antimicrobial activities exemplify that it can induce microbial cell death for all the strains at higher concentrations. MTT assay results portrayed the potent antiproliferative activity against MCF-7 cell lines (IC50 = 155.2 µg/ml) and non-cytotoxicity for MDA-MB-231 cell lines. Moreover, the synthesized resveratrol derivative induced ROS (reactive oxygen species) levels in MCF-7 cells, indicating cytotoxicity. CAXII gene expression study showed that it downregulated the CAXII genes. Conclusion: This study serves as an example of how a newly proposed resveratrol analogue might be utilized as a viable pharmacophore for specifically targeting the ER alpha protein which will be beneficial in investigating a fresh batch of effective resveratrol mimics as prospective anticancer agents.

From Tethered to Freestanding Stabilizers of 14‐3‐3 Protein‐Protein Interactions through Fragment Linking

AbstractSmall‐molecule stabilization of protein‐protein interactions (PPIs) is a promising strategy in chemical biology and drug discovery. However, the systematic discovery of PPI stabilizers remains a largely unmet challenge. Herein we report a fragment‐linking approach targeting the interface of 14‐3‐3 and a peptide derived from the estrogen receptor alpha (ERα) protein. Two classes of fragments—a covalent and a noncovalent fragment—were co‐crystallized and subsequently linked, resulting in a noncovalent hybrid molecule in which the original fragment interactions were largely conserved. Supported by 20 crystal structures, this initial hybrid molecule was further optimized, resulting in selective, 25‐fold stabilization of the 14‐3‐3/ERα interaction. The high‐resolution structures of both the single fragments, their co‐crystal structures and those of the linked fragments document a feasible strategy to develop orthosteric PPI stabilizers by linking to an initial tethered fragment.

From Tethered to Freestanding Stabilizers of 14-3-3 Protein-Protein Interactions through Fragment Linking.

Small-molecule stabilization of protein-protein interactions (PPIs) is a promising strategy in chemical biology and drug discovery. However, the systematic discovery of PPI stabilizers remains a largely unmet challenge. Herein we report a fragment-linking approach targeting the interface of 14-3-3 and a peptide derived from the estrogen receptor alpha (ERα) protein. Two classes of fragments-a covalent and a noncovalent fragment-were co-crystallized and subsequently linked, resulting in a noncovalent hybrid molecule in which the original fragment interactions were largely conserved. Supported by 20 crystal structures, this initial hybrid molecule was further optimized, resulting in selective, 25-fold stabilization of the 14-3-3/ERα interaction. The high-resolution structures of both the single fragments, their co-crystal structures and those of the linked fragments document a feasible strategy to develop orthosteric PPI stabilizers by linking to an initial tethered fragment.

Pre- and postnatal developmental exposure to the polychlorinated biphenyl mixture aroclor 1221 alters female rat pituitary gonadotropins and estrogen receptor alpha levels

Polychlorinated-biphenyls (PCBs) are industrial compounds, which were widely used in manufacturing of electrical parts and transformers. Despite being banned in 1979 due to human health concerns, they persist in the environment. In humans and experimental model systems, PCBs elicit toxicity in part by acting as endocrine-disrupting chemicals (EDCs). Aroclor 1221 (A1221) is a weakly estrogenic PCB mixture known to alter reproductive function in rodents. EDCs can impact hormone signaling at any level of the hypothalamic-pituitary-gonadal (HPG) axis, and we investigated the effects of A1221 exposure during the prenatal and postnatal developmental periods on pituitary hormone and steroid receptor expression in female rats. Examining offspring at 3 ages, postnatal day 8 (P8), P32 and P60, we found that prenatal exposure to A1221 increased P8 neonate pituitary luteinizing hormone beta (Lhb) mRNA and LHβ gonadotrope cell number while decreasing LH serum hormone concentration. No changes in pituitary hormone or hormone receptor gene expression were observed peri-puberty at P32. In reproductively mature rats at P60, we found pituitary follicle stimulating hormone beta (Fshb) mRNA levels increased by prenatal A1221 exposure with no corresponding alterations in FSH hormone or FSHβ expressing cell number. Estrogen receptor alpha (ERα) mRNA and protein levels were also increased at P60, but only following postnatal A1221 dosing. Together, these data illustrate that exposure to the PCB A1221, during critical developmental windows, alters pituitary gonadotropin hormone subunits and ERα levels in offspring at different phases of maturation, potentially impacting reproductive function in concert with other components of the HPG axis.

Cyclopia extracts act as selective estrogen receptor subtype downregulators in estrogen receptor positive breast cancer cell lines: Comparison to standard of care breast cancer endocrine therapies and a selective estrogen receptor agonist and antagonist.

Breast cancer is the most diagnosed type of cancer amongst women in economically developing countries and globally. Most breast cancers express estrogen receptor alpha (ERα) and are categorized as positive (ER+) breast cancer. Endocrine therapies such as, selective estrogen receptor modulators (SERMs), aromatase inhibitors (AIs), and selective estrogen receptor downregulators (SERDs) are used to treat ER+ breast cancer. However, despite their effectiveness, severe side-effects and resistance are associated with these endocrine therapies. Thus, it would be highly beneficial to develop breast cancer drugs that are as effective as current therapies, but less toxic with fewer side effects, and less likely to induce resistance. Extracts of Cyclopia species, an indigenous South African fynbos plant, have been shown to possess phenolic compounds that exhibit phytoestrogenic and chemopreventive activities against breast cancer development and progression. In the current study, three well characterized Cyclopia extracts, SM6Met, cup of tea (CoT) and P104, were examined for their abilities to modulate the levels of the estrogen receptor subtypes, estrogen receptor alpha and estrogen receptor beta (ERβ), which have been recognized as crucial to breast cancer prognosis and treatment. We showed that the Cyclopia subternata Vogel (C. subternata Vogel) extracts, SM6Met and cup of tea, but not the C. genistoides extract, P104, reduced estrogen receptor alpha protein levels while elevating estrogen receptor beta protein levels, thereby reducing the ERα:ERβ ratio in a similar manner as standard of care breast cancer endocrine therapies such as fulvestrant (selective estrogen receptor downregulator) and 4-hydroxytamoxifen (elective estrogen receptor modulator). Estrogen receptor alpha expression enhances the proliferation of breast cancer cells while estrogen receptor beta inhibits the proliferative activities of estrogen receptor alpha. We also showed that in terms of the molecular mechanisms involved all the Cyclopia extracts regulated estrogen receptor alpha and estrogen receptor beta protein levels through both transcriptional and translational, and proteasomal degradation mechanisms. Therefore, from our findings, we proffer that the C. subternata Vogel extracts, SM6Met and cup of tea, but not the C. genistoides extract, P104, selectively modulate estrogen receptor subtypes levels in a manner that generally supports inhibition of breast cancer proliferation, thereby demonstrating attributes that could be explored as potential therapeutic agents for breast cancer.

Codon adaptation by synonymous mutations impacts the functional properties of the estrogen receptor-alpha protein in breast cancer cells.

Oestrogen receptor-alpha (ERα) positivity is intimately associated with the development of hormone-dependent breast cancers. A major challenge in the treatment of these cancers is to understand and overcome the mechanisms of endocrine resistance. Recently, two distinct translation programmes using specific transfer RNA (tRNA) repertoires and codon usage frequencies were evidenced during cell proliferation and differentiation. Considering the phenotype switch of cancer cells to more proliferating and less-differentiated states, we can speculate that the changes in the tRNA pool and codon usage that likely occur make the ERα coding sequence no longer adapted, impacting translational rate, co-translational folding and the resulting functional properties of the protein. To verify this hypothesis, we generated an ERα synonymous coding sequence whose codon usage was optimized to the frequencies observed in genes expressed specifically in proliferating cells and then investigated the functional properties of the encoded receptor. We demonstrate that such a codon adaptation restores ERα activities to levels observed in differentiated cells, including: (a) an enhanced contribution exerted by transactivation function 1 (AF1) in ERα transcriptional activity; (b) enhanced interactions with nuclear receptor corepressor 1 and 2 [NCoR1 and NCoR2 (also known as SMRT) respectively], promoting repressive capability; and (c) reduced interactions with SRC proto-oncogene, non-receptor tyrosine kinase (Src) and phosphoinositide 3-kinase (PI3K) p85 kinases, inhibiting MAPK and AKT signalling pathway.

Protein-protein Docking Studies of Estrogen Receptor Alpha and TRIM56 Interaction for Breast Cancer Drug Screening

Breast cancer is the highest mortality cause in women with cancer. Protein-protein docking for target-based screening is an effective approach in breast cancer drug discovery via estrogen receptor (ER) signaling. TRIM56, an E3 ubiquitin protein ligase, can bind to and stabilize ER alpha. Thus, drug screening that can inhibit or weaken the interaction between ER alpha and TRIM56 is promising to obtain novel yet specific breast cancer drugs. In this study, we performed protein-protein docking studies for ER alpha and TRIM56 interaction and virtual screening for FDA-approved drugs from the ZINC database against ER alpha and TRIM56 complex protein model structure. We utilized Cluspro 2.0, PyRx 0.8, and Pymol 2.4.1 to conduct protein-protein docking, virtual screening, and model structure visualization. PIP and PLIP software were also applied to analyze the amino acid residue between proteins or protein-ligands. Based on the protein-protein docking, ER alpha and TRIM56 established interaction. Utilizing this complex protein as a macromolecule in the virtual screen of 1071 molecules of FDA-approved drugs, we obtain the top five lowest binding energy molecules i.e., dutasteride, dihydroergotamine, nilotinib, ergotamine, and bromocriptine. In addition, the energy binding affinity between ER alpha-dutasteride complex with TRIM56 was weakened in the presence of dutasteride. In conclusion, protein-protein docking between ER alpha-TRIM56 was able to select FDA-approved drugs that could bind to the complex, and dutasteride binding to ER alpha-TRIM56 complex weakened the interaction.Keywords: protein-protein docking, estrogen receptor alpha, TRIM56, breast cancer, ubiquitin.

Using Synthetic Biology methods to construct a functional estrogen biosensor based on the dimerization‐dependent Red Fluorescent Protein

The presence of estrogenic compounds (endocrine‐disruptors, EDCs) in the water supply raises concerns about human and aquatic health. Current methods for detecting estrogen contamination require expensive, time‐consuming techniques such as liquid chromatography‐mass spectrometry and high performance liquid chromatography. Previously reported estrogen biosensors required multiple cloning and transformation steps for successful detection in bacteria. Synthetic biology allows for the construction of genetic devises composed of DNA sequences modified to be interchangeable and provide novel functions. New tools and devices are constantly needed to enhance the already extensive list of novel genetic parts. Our approach to the design of an estrogen responsive element uses rational engineering to express dimerization dependent Red Fluorescent Proteins (ddRFP) fused to the Estrogen Receptor alpha protein (ERalpha) in bacteria cells. These two monomeric weakly fluorescent RFP proteins derived from mutant dTomato (Campbell et al,2021) are strongly fluorescent upon dimerization. These RFP constructs are brought together in the cell when ERalpha dimerizes in the presence of estrogen. We present here the construction strategy and proof of concept data demonstrating the efficiency of this dual‐functional biosensor and it effectiveness for detection of estrogenic compounds in contaminated water.

TRIM3 facilitates estrogen signaling and modulates breast cancer cell progression

BackgroundBreast cancer is the most common cancer in women worldwide. More than 70% of breast cancers are estrogen receptor (ER) alpha positive. Compared with ER alpha-negative breast cancer, which is more aggressive and has a shorter survival time, ER alpha-positive breast cancer could benefit from endocrine therapy. Selective estrogen receptor modulators, such as tamoxifen, are widely used in endocrine therapy. Approximately half of ER alpha-positive breast cancer patients will eventually develop endocrine resistance, making it a major clinical challenge in therapy. Thus, decoding the throughput of estrogen signaling, including the control of ER alpha expression and stability, is critical for the improvement of breast cancer therapeutics.MethodsTRIM3 and ER alpha protein expression levels were measured by western blotting, while the mRNA levels of ER alpha target genes were measured by RT–PCR. A CCK-8 assay was used to measure cell viability. RNA sequencing data were analyzed by Ingenuity Pathway Analysis. Identification of ER alpha signaling activity was accomplished with luciferase assays, RT–PCR and western blotting. Protein stability assays and ubiquitin assays were used to detect ER alpha protein degradation. Ubiquitin-based immunoprecipitation assays were used to detect the specific ubiquitination modification on the ER alpha protein.ResultsIn our current study, we found that TRIM3, an E3 ligase, can promote ER alpha signaling activity and breast cancer progression. TRIM3 depletion inhibits breast cancer cell proliferation and migration, while unbiased RNA sequencing data indicated that TRIM3 is required for the activity of estrogen signaling on the -genome-wide scale. The immunoprecipitation assays indicated that TRIM3 associates with ER alpha and promotes its stability, possibly by inducing K63-linked polyubiquitination of ER alpha. In conclusion, our data implicate a nongenomic mechanism by which TRIM3 stabilizes the ER alpha protein to control ER alpha target gene expression linked to breast cancer progression.ConclusionOur study provides a novel posttranslational mechanism in estrogen signaling. Modulation of TRIM3 expression or function could be an interesting approach for breast cancer treatment.Graphical abstract2noEgdU1b5aSV4mMoG9UypVideo abstract

Sexual Dimorphism in Small-intestinal Neuroendocrine Tumors: Lower Prevalence of Mesenteric Disease in Premenopausal Women.

ContextSmall-intestinal neuroendocrine tumors (SI-NETs) have a modest but significantly higher prevalence and worse prognosis in male patients.ObjectiveThis work aims to increase understanding of this sexual dimorphism in SI-NETs.Patients and MethodsRetrospectively, SI-NET patients treated in a single tertiary center were included and analyzed for disease characteristics. Estrogen receptor 1 (ESR1) and 2 (ESR2), progesterone receptor (PGR), and androgen receptor (AR) messenger RNA (mRNA) expression was assessed in primary tumors and healthy intestine. Estrogen receptor alpha (ERα) and AR protein expression were analyzed by immunohistochemistry in primary tumors and mesenteric metastases.ResultsOf the 559 patients, 47% were female. Mesenteric metastasis/fibrosis was more prevalent in men (71% / 46%) than women (58% / 37%; P = 0.001 and P = 0.027, respectively). In women, prevalence of mesenteric metastases increased gradually with age from 41.1% in women <50 years to 71.7% in women >70 years. Increased expression of ESR1 and AR mRNA was observed in primary tumors compared to healthy intestine (both P < 0.001). ERα staining was observed in tumor cells and stroma with a strong correlation between tumor cells of primary tumors and mesenteric metastases (rho = 0.831, P = 0.02), but not in stroma (rho = −0.037, P = 0.91). AR expression was only found in stroma.ConclusionSexual dimorphism in SI-NETs was most pronounced in mesenteric disease, and the risk of mesenteric metastasis in women increased around menopause. The combination of increased ERα and AR expression in the SI-NET microenvironment suggests a modulating role of sex steroids in the development of the characteristic SI-NET mesenteric metastasis and associated fibrosis.

In Silico Approaches for the Identification of Novel Inhibitors against Breast Cancer Up-Regulated Protein

Breast cancer is a type of cancer that develops in the breast tissues. When some breast cells begin to grow abnormally, breast cancer develops. These cells grow and divide at a faster rate than healthy cells and continue to grow, generating a lump or mass. Cancer cells in the breast may spread to lymph nodes or other places of the body. The hormone estrogen encourages cancer growth when it binds to the receptor of the targeted protein. The purpose of this study is the rational screening of a 15,000 phytochemicals library against the estrogen receptor alpha protein. The library was employed for molecular docking to find the binding affinities and simulation analysis of the top-selected compounds. The top four compounds, Mangostenone E, Exiguaflavanone M, Sanggenon A, and Flaccidine were identified as direct inhibitors of estrogen receptors as evident from their high binding affinity and occupancy of specific binding sites. Mangostenone E was the leading phytochemical that showed a high docking score—15.97 (kcal/mol)—and bonding interaction at the active site of Mangostenone E. Leading phytochemicals were subjected to analysis for drug-like properties that further reinforced their validation. Potential molecules identified in this study can be considered lead drugs for the treatment of breast cancer. KEYWORDS Bioinformatics, docking, drug candidates, molecular dynamic simulation, phytochemicals, protein data bank

An Efficient One Pot Multicomponent Synthesis of Dihydro Pyrazolyl Bisthiazole Derivatives as Potential Anticancer Agents and their Molecular Docking Studies

A novel series of dihydropyrazolyl bisthiazoles were synthesized through simple reaction conditions via one pot multicomponent reaction of 2-bromo-1-(4-methyl-2-phenyl thiazol-5-yl)ethan-1-one, thiosemicarbazide and chalcones in the presence of sodium hydroxide in ethanol under reflux condition. The reaction generates two potential five membered heterocylic pharmacophores i.e. Hantzsch thiazole and dihydropyrazole in one step through simple operation, in shorter reaction time with high yields. The newly synthesized compounds were well characterized by IR, 1H, 13C NMR and mass spectral data. All the newly synthesized compounds were evaluated for their anticancer activity against human cancer cell lines and calculated their binding energy values with respect to 3ert protein. Some of the compounds exhibited excellent activity against MCF-7 cancer cell line and studied molecular interaction of probable target protein, human estrogen receptor alpha protein (3ert.pdb) using docking simulation.