Exploring the Effects of Endocrine-Disrupting Chemicals and miRNA Expression in the Pathogenesis of Endometriosis by Unveiling the Pathways: a Systematic Review.

Endometriosis, characterized by endometrial-like mucosal tissue outside the uterine cavity, is a reproductive disorder afflicting about 10% of women within the reproductive age. The pathogenesis of endometriosis has been attributed to factors like genetics, environmental particles, and hormones. A comprehensive review of studies from July 2010 to July 2023 across multiple databases was done to aid in a better understanding of the same. The investigation focused on studies delineating the correlation between endocrine disruptors, microRNAs, and endometriosis. To optimize the search scope, keywords and subject headings were used as search terms. Then, two authors rigorously assessed studies using criteria, selecting 27 studies from various databases. Notably, dioxins, organochlorine pesticides, and polychlorinated biphenyls exhibited a solid connection for endometriosis, while bisphenol A and phthalates yielded conflicting results. The heightened presence of bisphenol A, polychlorinated biphenyls, and phthalates was linked to altered gene expression, including genes like AKR1B10, AKR1C3, and FAM49B. MicroRNAs like miRNA-31, miRNA-144, and miRNA-145 emerged as vital factors in the onset of endometriosis and progression. Furthermore, elevated expression of miR-1304-3p, miR-544, and miR-3684 and reduced expression of miR-3935 and miR-4427 exert substantial influence on signaling pathways like NF-κB, MAPK, and Wnt/ß-catenin. Currently, literature shows an independent link between endocrine disruptor exposure and endometriosis and between microRNA dysregulation and endometriosis. However, research lacks the combination of all three factors. The review delves into the effects of endocrine disruptors and microRNAs on the pathogenesis of endometriosis to improve our understanding of the disorder and in finding therapies.

Prenatal bisphenol-A exposure altered exploratory and anxiety-like behaviour and induced non-monotonic, sex-specific changes in the cortical expression of CYP19A1, BDNF and intracellular signaling proteins in F1 rats.

Bisphenol-A (BPA) is one of the extensively studied estrogenic endocrine disrupting chemicals (EDC) with ubiquitous exposure among humans and wildlife. While there are literature reporting the association of dysregulated Brain-derived neurotrophic factor (BDNF) expression levels with altered cognitive and emotional behaviour such as anxiety-like and stress behaviour in animal models, there are no studies in BPA that investigate these altered neurobehavioural outcomes in parallel with the expression of intracellular proteins involved in BDNF signaling pathway. In this study, pregnant Wistar rats were exposed to BPA through water (25 µg/L, 250 µg/L, and 2.5 mg/L) during gestation day (GD) 9-21. Prenatal BPA exposure, increased anxiety-like behaviour in males and decreased exploratory behaviour in both male and female offspring. Downregulation of both BDNF and CYP19A1 genes were observed in male BPA-exposed offspring, whereas in females, the expression was upregulated. The expression of p-AKT, p-MEK and p-ERK proteins were increased in males, while in females, it decreased. Both the male and the female BPA-exposed offspring exhibited elevated levels of DNMT1 protein. The sex-specific alteration in the expression of CYP19A1 and DNA methyltransferase 1 (DNMT1) suggests that both hormonal and epigenetic dysregulation could underlie the long-term BPA-induced effect on anxiety-like behaviour in the offspring.

Prenatal exposure to bisphenol-A altered miRNA-224 and protein expression of aromatase in ovarian granulosa cells concomitant with elevated serum estradiol levels in F1 adult offspring.

This study was aimed to predict bisphenol-A (BPA)-responsive miRNA's using an in silico approach and to study their expression in granulosa cells of animals exposed prenatally to BPA. Pregnant Wistar rats were exposed to BPA through water (25 µg/L, 250 µg/L, and 2.5 mg/L) during gestation. The expression of miRNA-133b, miRNA-378 and miRNA-224 were analyzed in ovarian granulosa cells. BPA affected the postnatal developmental landmarks such as weight of the pups at birth and reduced anogenital distance. BPA exposed animals showed elevated serum estradiol (E2) levels, while follicle-stimulating hormone levels were reduced. The expression of miRNA-224 and aromatase protein levels were found to be increased. This preliminary finding reveals the impact of early life exposure to BPA on the long-term ovarian functions that may be mediated through miRNA-based granulosa cell response. Besides, it is also a compelling indicator for the subclinical response that could have important consequences on female fertility.

Transient exposure of methylparaben to zebrafish (Danio rerio) embryos altered cortisol level, acetylcholinesterase activity and induced anxiety-like behaviour.

Parabens are widely used as antimicrobial preservatives. Recent studies have reported the endocrine disrupting effects of these chemicals, especially methylparaben. Previously, we have reported the alteration in Vtg gene expression upon exposure to environmentally relevant doses of methylparaben in zebrafish (Danio rerio) embryos. However, studies reporting neurobehavioural outcomes on exposure to methylparaben are limited. Therefore, this study was aimed at investigating the methylparaben-induced effects on developmental and neurobehavioural endpoints. Zebrafish embryos were exposed to sub-lethal concentrations of methylparaben: 0.1 ppb, 1 ppb, 10 ppb and 100 ppb. Alterations in developmental landmarks such as heart rate and hatching percentage were observed in embryos exposed to 10 ppb and 100 ppb of methylparaben. Results obtained from the novel tank diving test established that anxiety-like behaviour is induced in larvae exposed to 0.1 ppb and 1 ppb of methylparaben. A significant inhibition in the acetylcholinesterase (AChE) activity was also recorded in methylparaben-exposed groups. An increase in cortisol levels was observed in the exposed groups, which further supports the observations made in the novel tank diving test, establishing methylparaben as an anxiogenic agent even at sub-lethal concentrations. The underlying molecular mechanism needs further elucidation to investigate whether the behavioural effects are proximally or distally induced by early developmental exposure to methylparaben.

Developmental toxicity and induction of vitellogenin in embryo-larval stages of zebrafish (Danio rerio) exposed to methyl Paraben.

MethylParaben (MP), a methyl ester of p-hydroxybenzoic acid, is used as an anti-microbial preservative in foods, drugs and cosmetics for decades. It enters the aquatic environment, and can have toxic effects on aquatic organisms. Little is known on the developmental toxicity of MP exposure to zebrafish during early life stages. In this study, the developmental effects of MP were evaluated in embryo-larval zebrafish (at concentrations ranging from 100µM, 200µM, 400µM, 800µM and 1000µM for 96h post fertilization (hpf). The survival, hatching, heart beat rate and developmental abnormalities were observed in the embryos exposed to MP. MP exposure resulted in decreased heart rate and hatching rate. Defects including pericardial edema blood cell accumulation and bent spine were observed in all the treated concentration, except at 100µM. With increasing concentrations, the frequency of these defects increased. The 96 hpf LC50 of MP was calculated to be 428µM (0.065mg/L). Furthermore, RT-PCR result showed that in larval zebrafish exposed to 100µM (0.015mg/L) of MP till 96 hpf, expression of vitellogenin I (Vtg -I) was significantly upregulated compared to the control group. This data suggest that even though lower concentrations of MP do not cause phenotypic malformations, it leads to dysregulated expression of estrogenic biomarker gene Vtg-I.

Dimorphic placental stress: A repercussion of interaction between endocrine disrupting chemicals (EDCs) and fetal sex.

Placental homeostasis is critical for fetal development as it determines the health of mother and fetus during pregnancy and in later life. Interestingly even the fetus, in a sexually dimorphic manner, influences the pedantic growth and development of placenta. Although placenta is thought to act as a protective barrier against chemical exposures, certain endocrine disrupting chemicals (EDCs) that are circulating in mother's blood tend to cross placenta. These EDCs have been reported to cause changes in expression levels of certain genes, immunogenic factors and non-coding RNAs such as micro RNA (miRNA) and long non-coding RNA (lncRNA) leading to placental stress. We hypothesize that these changes in placenta occur in a sexually dimorphic manner as a result of interaction between EDC exposure and fetal sex. Therefore, we propose that the ability of placenta to respond and buffer EDC exposure depends on fetal sex and, hence the EDC associated disease susceptibility of one sex differs from the other.

Detection of phenolic endocrine disrupting chemicals (EDCs) from maternal blood plasma and amniotic fluid in Indian population.

There is a widespread exposure of general population, including pregnant women and developing fetuses, to the endocrine disrupting chemicals (EDCs). These chemicals have been reported to be present in urine, blood serum, breast milk and amniotic fluid. We aimed to investigate the association between the maternal exposure and in utero fetal exposure levels of these chemicals to study their transfer from maternal to fetal unit indicating prenatal exposure. Samples of maternal blood and amniotic fluid were collected as set from 53 pregnant women at full term. Nine phenolic EDCs, methyl paraben (MP; 20.92ng/mL and 18.92ng/mL), ethyl paraben (EP; 1.97ng/ mL and 1.89ng/mL), propyl paraben (PP; 19.22ng/mL and 18.82ng/mL), butyl paraben (BP; 1.11ng/mL and 1.37ng/mL), p-hydroxybenzoic acid (PHBA; 29.99ng/mL and 26.15ng/mL), bisphenol A (BPA; 7.43ng/mL and 7.75ng/mL), triclosan (TCS; 7.17ng/mL and 7.04ng/mL), octyl phenol (OP; 5.46ng/mL and 5.72ng/mL) and nonyl phenol (NP; 9.38ng/mL and 8.44ng/mL), were simultaneously detected in samples of maternal blood plasma and amniotic fluid respectively using Gas Chromatography-Mass Spectrometry (GC-MS). Highest positive correlation was found for total concentration of 4-nonyl phenol, NP (r=0.575, p<0.001), whereas the lowest positive correlation was found for free form of bisphenol A, BPA (r=0.343, p<0.05), when compared between the two matrices. Our results suggest that maternal exposure to several EDCs is positively associated with in utero exposure to the developing fetus. Future studies should focus on collection of amniotic fluid at different trimesters and the corresponding maternal samples to better characterize the pharmacokinetics and the associated disease etiologies of these EDCs during fetal development.

Systems biological approach on neurological disorders: a novel molecular connectivity to aging and psychiatric diseases.

BACKGROUND: Systems biological approach of molecular connectivity map has reached to a great interest to understand the gene functional similarities between the diseases. In this study, we developed a computational framework to build molecular connectivity maps by integrating mutated and differentially expressed genes of neurological and psychiatric diseases to determine its relationship with aging. RESULTS: The systematic large-scale analyses of 124 human diseases create three classes of molecular connectivity maps. First, molecular interaction of disease protein network generates 3632 proteins with 6172 interactions, which determines the common genes/proteins between diseases. Second, Disease-disease network includes 4845 positively scored disease-disease relationships. The comparison of these disease-disease pairs with Medical Subject Headings (MeSH) classification tree suggests 25% of the disease-disease pairs were in same disease area. The remaining can be a novel disease-disease relationship based on gene/protein similarity. Inclusion of aging genes set showed 79 neurological and 20 psychiatric diseases have the strong association with aging. Third and lastly, a curated disease biomarker network was created by relating the proteins/genes in specific disease contexts, such analysis showed 73 markers for 24 diseases. Further, the overall quality of the results was achieved by a series of statistical methods, to avoid insignificant data in biological networks. CONCLUSIONS: This study improves the understanding of the complex interactions that occur between neurological and psychiatric diseases with aging, which lead to determine the diagnostic markers. Also, the disease-disease association results could be helpful to determine the symptom relationships between neurological and psychiatric diseases. Together, our study presents many research opportunities in post-genomic biomarkers development.

Metallomic profiling and linkage map analysis of early Parkinson's disease: a new insight to aluminum marker for the possible diagnosis.

BACKGROUND: Parkinson's disease (PD) is the most common neurodegenerative disorder. The diagnosis of PD is challenging and currently none of the biochemical tests have proven to help in diagnosis. Serum metallomic analysis may suggest the possibility of diagnosis of PD. METHODOLOGY/RESULTS: The metallomic analysis was targeted on 31 elements obtained from 42 healthy controls and 45 drug naive PD patients using ICP-AES and ICP-MS to determine the concentration variations of elements between PD and normal. The targeted metallomic analysis showed the significant variations in 19 elements of patients compared to healthy control (p<0.04). The partial least squares discriminant analysis (PLS-DA) showed aluminium, copper, iron, manganese and zinc are the key elements, contributes the separation of PD patients from control samples. The correlation coefficient analysis and element-element ratio confirm the imbalance of inter-elements relationship in PD patients' serum. Furthermore, elements linkage map analysis showed aluminium is a key element involved in triggering of phosphorus, which subsequently lead to imbalance of homeostatic in PD serum. The execution of neural network using elements concentrations provides 95% accuracy in detection of disease. CONCLUSIONS/SIGNIFICANCE: These results suggest that there is a disturbance in the elements homeostasis and inter-elements relationship in PD patients' serum. The analysis of serum elements helps in linking the underlying cellular processes such as oxidative stress, neuronal dysfunction and apoptosis, which are the dominating factors in PD. Also, these results increase the prospect of detection of early PD from serum through neural network algorithm.

Metabolic profiling of Parkinson's disease: evidence of biomarker from gene expression analysis and rapid neural network detection.

BACKGROUND: Parkinson's disease (PD) is a neurodegenerative disorder. The diagnosis of Parkinsonism is challenging because currently none of the clinical tests have been proven to help in diagnosis. PD may produce characteristic perturbations in the metabolome and such variations can be used as the marker for detection of disease. To test this hypothesis, we used proton NMR and multivariate analysis followed by neural network pattern detection. METHODS & RESULTS: 1H nuclear magnetic resonance spectroscopy analysis was carried out on plasma samples of 37 healthy controls and 43 drug-naive patients with PD. Focus on 22 targeted metabolites, 17 were decreased and 5 were elevated in PD patients (p < 0.05). Partial least squares discriminant analysis (PLS-DA) showed that pyruvate is the key metabolite, which contributes to the separation of PD from control samples. Furthermore, gene expression analysis shows significant (p < 0.05) change in expression of PDHB and NPFF genes leading to increased pyruvate concentration in blood plasma. Moreover, the implementation of 1H- NMR spectral pattern in neural network algorithm shows 97.14% accuracy in the detection of disease progression. CONCLUSION: The results increase the prospect of a robust molecular definition in detection of PD through the early symptomatic phase of the disease. This is an ultimate opening for therapeutic intervention. If validated in a genuinely prospective fashion in larger samples, the biomarker trajectories described here will go a long way to facilitate the development of useful therapies. Moreover, implementation of neural network will be a breakthrough in clinical screening and rapid detection of PD.