Estrogens Regulate Placental Angiogenesis in Horses

A sufficient vascular network within the feto-maternal interface is necessary for placental function. Several pregnancy abnormalities have been associated with abnormal vascular formations in the placenta. We hypothesized that growth and expansion of the placental vascular network in the equine (Equus caballus) placenta is regulated by estrogens (estrogen family hormones), a hormone with a high circulating concentration during equine gestation. Administration of letrozole, a potent and specific inhibitor of aromatase, during the first trimester (D30 to D118), decreased circulatory estrone sulfate concentrations, increased circulatory testosterone and androstenedione concentrations, and tended to reduce the weight of the fetus (p < 0.1). Moreover, the gene expression of CYP17A1 was increased, and the expression of androgen receptor was decreased in the D120 chorioallantois (CA) of letrozole-treated mares in comparison to that of the control mares. We also found that at D120, the number of vessels tended to decrease in the CAs with letrozole treatment (p = 0.07). In addition, expression of a subset of angiogenic genes, such as ANGPT1, VEGF, and NOS2, were altered in the CAs of letrozole-treated mares. We further demonstrated that 17β-estradiol increases the expression of ANGPT1 and VEGF and increases the angiogenic activity of equine endothelial cells in vitro. Our results from the estrogen-suppressed group demonstrated an impaired placental vascular network, suggesting an estrogen-dependent vasculogenesis in the equine CA during the first trimester.

In the Model Cell Lines of Moderately and Poorly Differentiated Endometrial Carcinoma, Estrogens Can Be Formed via the Sulfatase Pathway

Endometrial cancer (EC) is the most common gynecological malignancy in resource-abundant countries. The majority of EC cases are estrogen dependent but the mechanisms of estrogen biosynthesis and oxidative metabolism and estrogen action are not completely understood. Here, we evaluated formation of estrogens in models of moderately and poorly differentiated EC: RL95-2 and KLE cells, respectively. Results revealed high expression of estrone-sulfate (E1-S) transporters (SLCO1A2, SLCO1B3, SLCO1C1, SLCO3A1, SLC10A6, SLC22A9), and increased E1-S uptake in KLE vs RL95-2 cells. In RL95-2 cells, higher levels of sulfatase and better metabolism of E1-S to E1 were confirmed compared to KLE cells. In KLE cells, disturbed balance in expression of HSD17B genes led to enhanced activation of E1 to E2, compared to RL95-2 cells. Additionally, increased CYP1B1 expression and down-regulation of genes encoding phase II metabolic enzymes: COMT, NQO1, NQO2, and GSTP1 suggested decreased detoxification of carcinogenic metabolites in KLE cells. Results indicate that in model cell lines of moderately and poorly differentiated EC, estrogens can be formed via the sulfatase pathway.

Steroid sulfatase deficiency causes cellular senescence and abnormal differentiation by inducing Yippee-like 3 expression in human keratinocytes

Human steroid sulfatase (STS) is an enzyme that catalyzes the hydrolysis of dehydroepiandrosterone sulfate (DHEAS), estrone sulfate (E1S), and cholesterol sulfate. Abnormal expression of STS causes several diseases including colorectal, breast, and prostate cancer and refractory skin disease. In particular, accumulation of intracellular cholesterol sulfate by STS deficiency leads to a skin disorder with abnormal keratinization called X-linked ichthyosis (XLI). To determine the detailed mechanisms of XLI, we performed RNA sequencing (RNA-seq) analysis using human keratinocyte HaCaT cells treated with cholesterol and cholesterol sulfate. Of the genes with expression changes greater than 1.5-fold, Yippee-like 3 (YPEL3), a factor expected to affect cell differentiation, was found. Induction of YPEL3 causes permanent growth arrest, cellular senescence, and inhibition of metastasis in normal and tumor cells. In this study, we demonstrate that YPEL3 expression was induced by STS deficiency and, using the CRISPR/Cas9 system, a partial knock-out (STS+/−) cell line was constructed to establish a disease model for XLI studies. Furthermore, we show that increased expression of YPEL3 in STS-deficient cell lines promoted cellular senescence and expression of keratinization-related proteins such as involucrin and loricrin. Our results suggest that upregulation of YPEL3 expression by STS deficiency may play a crucial role in inducing cellular senescence and abnormal differentiation in human keratinocytes.

Organic Anion Transporting Polypeptide 2B1 (OATP2B1) Genetic Variants: In Vitro Functional Characterization and Association With Circulating Concentrations of Endogenous Substrates

Organic anion transporting polypeptide 2B1 (OATP2B1, gene SLCO2B1) is an uptake transporter that is thought to determine drug disposition and in particular, the oral absorption of medications. At present, the clinical relevance of SLCO2B1 genetic variation on pharmacokinetics is poorly understood. We sought to determine the functional activity of 5 of the most common missense OATP2B1 variants (c.76_84del, c.601G&gt;A, c.917G&gt;A, c.935G&gt;A, and c.1457C&gt;T) and a predicted dysfunctional variant (c.332G&gt;A) in vitro. Furthermore, we measured the basal plasma concentrations of endogenous OATP2B1 substrates, namely estrone sulfate, dehydroepiandrosterone sulfate (DHEAS), pregnenolone sulfate, coproporphyrin I (CPI), and CPIII, and assessed their relationships with SLCO2B1 genotypes in 93 healthy participants. Compared to reference OATP2B1, the transport activities of the c.332G&gt;A, c.601G&gt;A and c.1457C&gt;T variants were reduced among the substrates examined (estrone sulfate, DHEAS, CPI, CPIII and rosuvastatin), although there were substrate-dependent effects. Lower transport function of OATP2B1 variants could be explained by diminished cell surface expression. Other OATP2B1 variants (c.76-84del, c.917G&gt;A and c.935G&gt;A) had similar activity to the reference transporter. In the clinical cohort, the SLCO2B1 c.935G&gt;A allele was associated with both higher plasma CPI (42%) and CPIII (31%) concentrations, while SLCO2B1 c.917G&gt;A was linked to lower plasma CPIII by 28% after accounting for the effects of age, sex, and SLCO1B1 genotypes. No association was observed between SLCO2B1 variant alleles and estrone sulfate or DHEAS plasma concentrations, however 45% higher plasma pregnenolone sulfate level was associated with SLCO2B1 c.1457C&gt;T. Taken together, we found that the impacts of OATP2B1 variants on transport activities in vitro were not fully aligned with their associations to plasma concentrations of endogenous substrates in vivo. Additional studies are required to determine whether circulating endogenous substrates reflect OATP2B1 activity.

Association between the Area of the Highest Flank Temperature and Concentrations of Reproductive Hormones during Pregnancy in Polish Konik Horses—A Preliminary Study

Determination of the pregnancy status is one of the most important factors for effective pregnancy management. Knowledge of the stage of pregnancy is important to interpret many of the reproductive hormones’ concentrations, including progesterone (P4), estrone sulfate (E1S), 17-ß estradiol (E2), and relaxin (REL). However, it is limited in wildlife or captive equids that cannot be handled. Reproductive hormones affect regional blood flow, the proliferation of tissues, and local metabolism intensity. Therefore, this preliminary study aimed to assess changes in thermal features of the abdomen lateral surface and concentrations of reproductive hormones in Polish native pregnant mares. The study was carried out on 14 non-pregnant and 26 pregnant Polish Konik mares during eleven months of pregnancy. Infrared thermography was conducted to image the lateral surface of mares' abdomen (Px1) and flank area (Px2); P4, E1S, E2, and REL concentrations in serum were also determined. The evidence of the association between the area with the highest temperatures (Area of Tmax) and serum concentrations of P4 (the slope = 1.373; p = 0.9245) and REL (the slope = 1.342; p = 0.4324) were noted dependent across months of pregnancy. Measures of superficial body temperatures were found to change monthly, similarly to ambient temperatures, with no evidence of coincidence with changes in reproductive hormone concentrations. Individual thermal characteristics of the lateral surface of the abdomen differed between pregnant and non-pregnant mares in other periods. Differences in maximal and average temperature and Area of Tmax were observed from the sixth month of pregnancy, and those in minimal temperature were observed from the eighth month.

Dual Aromatase-Sulphatase Inhibitors (DASIs) for the Treatment of Hormone Dependent Breast Cancer

: Breast cancer is the most frequent diagnosed cancer in women and the second most common form of cancer, causing death after lung cancer, all across the globe at an alarming rate. The level of estrogens, in breast cancer tissues of postmenopausal women is 10-40 folds higher than the non-carcinogenic breast tissues. As a result of this greater level of estrogen, breast tissue becomes more prone to develop breast cancer; mainly estradiol plays a significant role in the initiation and development of hormone dependent breast cancer. Androstenedione, Adrenal dehydroepiandrosterone sulfate and estrone-sulfate also plays an important role of precursor for estrogen biosynthesis. Estrogens deprivation exhibits an attractive phenomena in the advancement of most ideal therapeutics for the treatment of breast cancer. Inhibition of aromatase and sulphatase emerged as attractive therapy for the treatment of hormone dependent breast cancer via deprivation of estrogen by different pathways. The cocktail of aromatase and sulphatase inhibitors known as dual aromatase-sulphatase inhibitors (DASIs) emerged as an attractive approach for the effective estrogen deprivation. The present review article focused on the journey of dual aromatase-sulphatase inhibitors from the beginning to till date (2020). Keeping in view the key observations, this review may be helpful for medicinal chemists to design and develop new and efficient dual aromatase-sulphatase inhibitors for the possible treatment of hormone-related breast cancer.

PSI-30 Evaluation of urine estrone sulfate (P-Test) in beef cattle as an alternative pregnancy detection method

Pregnancy detection in cattle can be expensive and labor intensive to producers. A simple, cost-efficient method of determining early pregnancy, chute side, is lacking within the industry. This study tested various proven pregnancy detection methods against an alternative method. Pregnancy was assessed by ultrasonography, blood serum pregnancy-specific protein B (PSPB), and urine estrone sulfate (“P-Test”). A fixed timed AI (FTAI) protocol was conducted on 14 predominantly British cattle housed at Tarleton State University in Stephenville, TX. Cows were synchronized using the co-synch plus 7 day CIDR protocol, with PGF injection of d 7. Heifers were synchronized similarly with the exception of GnRH injection on d 1. Estrotech patch scores (1 – 4, 1 showing little signs of estrus to 4 presenting signs of estrus) were utilized to indicate estrus activity prior to FTAI (21.4% vs. 78.6% females scored 1 vs. 4, respectively). FTAI was performed on protocol d 10 along with a 2mL injection of GnRH. Serum for PSPB analysis was collected at d 30 and 60 post insemination, while urine was sampled on d 60. Overall 53.8% (ultrasonography) and 71.4% (PSPB) of cattle were determined to be pregnant at d 30 post FTAI. Chi squared analysis proved that PSPB and ultrasound were similar at detecting pregnancy (P = 0.04). At d 60 “P-Test” color varied indicating differing levels of estrone sulfate. A t-test revealed increased PSPB from d 30–60 post FTAI (P &lt; 0.0001); however, “P-Test” could not predict similar results (P = 0.655). Data suggest the “P-Test” is not a consistent method to diagnose early pregnancy. Future studies may be warranted to investigate a more desirable time point for utilizing the chute side “P-Test” as a reliable method to determine pregnancy.