Gonadotropin Research Articles

A synthetic cyclic peptide for promoting antigen presentation and immune activation

Cyclic peptides are often used as scaffolds for the multivalent presentation of drug molecules due to their structural stability and constrained conformation. We identified a cyclic deca-peptide incorporating lipoamino acids for delivering T helper and B cell epitopes against group A Streptococcus (GAS), eliciting robust humoral immune responses. In this study, we assessed the function-immunogenicity relationship of the multi-component vaccine candidate (referred to as VC-13) to elucidate a mechanism of action. We identified a potential universal delivery platform, not only capable of adjuvanting different peptide epitopes (e.g., NS1 and 88/30 from group A Streptococcus, gonadotropin hormone releasing hormone [GnRH]), but also protein antigens (e.g., bovine serum albumin [BSA], receptor binding domain (RBD) of the SARS-CoV-2 protein responsible for COVID-19 infection [SARS-CoV-2 RBD]) and small molecular haptens (e.g., cocaine). All vaccine candidates self-assembled into sub-500 nm nanoparticles and induced high antigen-specific systemic IgG titers and opsonic potential compared to the antigen co-administered with a commercial adjuvant, complete Freund’s adjuvant. Notably, presence of the cyclic decapeptide in this vaccine increased accumulation in the draining inguinal lymph nodes, facilitating cellular uptake of peptide antigens. Furthermore, the lipoamino acid promoted dendritic cell activation, acting as both toll-like receptors 2 and 4 -targeting moiety. Our study revealed the importance of the cyclic decapeptide and lipoamino acid presence in antigen presentation and immune response activation, leading onto the development of a fully synthetic, self-assembled, and promising platform for the delivery of subunit vaccines and anti-drug vaccines.

Effect of administration of nanoparticles of human chorionic gonadotropin on testicular hemodynamics, testicular volume, testicular echotexture, and circulating testosterone and nitric oxide in pubescent goat bucks under heat stress conditions.

The current study investigated the effect of a single administration of human chorionic gonadotropin hormone (hCG) and its nanoparticles (NPs) on testicular hemodynamics using Doppler ultrasonography, testicular volume, testicular echotexture (PIX), and circulating testosterone and nitric oxide (NO) in pubescent goat bucks during summer months. Fifteen Baladi goats were divided into three groups (5 in each) and subjected to a single intramuscular administration of one ml of physiological saline ( control group), one ml containing 500 IU of hCG (hCG group) or one ml containing 125 IU of hCG NPs (hCG NPs group). Testicular hemodynamics assessment was done just before administration (0h), and at 2, 4, 6, 24, and daily till 7 days after administration. Testicular volume and PIX were measured and analyzed by B-mode ultrasonography and computer analysis software. Testosterone and NO concentrations were measured using commercial kits. Results revealed significant decreases (P ˂0.05) in Doppler indices values (resistance index and pulsatility index) especially at 24h in the hCG group (0.314 ± 0.01, 0.382 ± 0.01, respectively) compared with the hCG NPs group (0.428 ± 0.01, 0.567 ± 0.02, respectively), and the control group (0.464 ± 0.006, 0.64 ± 0.01, respectively). There were significant increases (P < 0.001) in testicular volume, PIX, testosterone, and NO concentrations in hCG NPs and hCG groups compared to the control group. There were no significant differences between the hCG NPs and hCG groups (P ˃ 0.05) in testicular volume and PIX, while testosterone significantly increased in the hCG group compared to the hCG NPs group and NO was significantly increased in the hCG NPs group compared to the hCG group. In conclusion, a single administration of hCG or its NPs improved testicular vascularization, testicular volume, and circulating testosterone and NO in pubescent goat bucks. So, hCG NPs could be recommended for improving the reproductive performance of goat bucks under heat-stress conditions.

Gonadotropin therapy for mini-puberty induction in male infants with hypogonadotropic hypogonadism: a systematic review.

Congenital hypogonadotropic hypogonadism (CHH) is defined as an isolated deficiency of gonadotropin hormones. Mini-puberty, a transient postnatal activation of the hypothalamic-pituitary-gonadal axis in healthy infants, provides a window of opportunity to diagnose and treat CHH. Currently, in male infants with CHH, testosterone is used to increase phallus size. However, gonadotropin replacement could additionally promote testicular descent and development, particularly relating to Sertoli cells. We conducted a systematic review of the effectiveness of gonadotropin therapy in stimulating mini-puberty related outcomes in male infants with CHH. In line with PRISMA guidelines, a systematic review of eleven databases was carried out (August 2023). Evidence quality was assessed using the Cochrane Risk of Bias for Non-Randomised Studies of Interventions tool. Protocol registered on PROSPERO (CRD42023453080). After a double-consensus screen of 767 abstracts and 66 full texts, 11 studies were included from seven countries. 71 male infants were enrolled, 12 with Kallmann syndrome. Median age at treatment initiation was 4.2 months (0.25-57) and follow-up ranged from 3 to 10 years. Gonadotropin therapy was administered using continuous subcutaneous infusion (n=35) or subcutaneous injection (n=36). Due to treatment variability, modalities were combined for data synthesis. Gonadotropins induced a significant increase in penile length and inhibin B concentration (P=0.0007) and led to partial or full testicular descent in 73% (n=62) of patients. This systematic review provides unique evidence supporting the efficacy of gonadotropins for induction of mini-puberty. However, the reliability and generalisability are limited due to disparate data and treatment modality variation.

ULTRASOUND APPEARANCE OF THE ENDOMETRIUM AND CLINICAL CORRELATION IN A CASE OF AMENORRHEA

The subject of this paper is the presentation of a clinical case of amenorrhea in a young woman with regular menstrual cycles and a positive pregnancy test. The ultrasound examination of the uterus at 5 weeks and 1 day after the last menstruation showed the absence of a gestational sac and a thin and hypoechoic endometrium. Since the pregnancy test was positive, the ultrasound examination was repeated after 7 days. During this interval, the patient presented a hemorrhage. The ultrasound performed 4 days after showed a thin endometrium with medium echogenicity. Repeating at 48 hours the measurement of the human chorionic gonadotrophin hormone showed low values that decreased, which correlated with the clinical and ultrasound aspects confirmed the diagnosis of early pregnancy failure.

Clinical and Analytical Presentation of Central Precocious Puberty: A 20-Year Cross-Sectional Study

Objectives − This study aims to compare the clinical and analytical presentation of Central precocious puberty (CPP), considering age.Methods − An observational, cross-sectional study was conducted on children diagnosed with CPP at a level III hospital, between January 2002 and April 2022. Clinical, auxological, sociodemographic, laboratory, and imaging parameters were analyzed.Results − Out of the 52 children studied, the majority were girls (N=44). The median age of puberty onset in girls was 6.79 years and the mean age at first consultation of 8.13 years, with a significantly lower age at hospital referral (7.65 years; P=0.045) compared to boys. Idiopathic etiology was predominant in both. In girls, breast development appeared at older mean ages (P=0.009), while pubic hair growth and accelerated growth were associated with younger ages at puberty onset (P=0.021; P=0.018, respectively). Basal and peak levels of gonadotropin hormones were higher in girls, although not statistically significant. In girls, age at puberty onset correlated negatively with standard deviation of body mass index (P=0.023), while age at first consultation correlated positively with bone age (P&lt;0.001), and was associated with younger ages at Gonadotropin-Releasing Hormone stimulation test (P=0.020).Conclusion − This study provides innovative and relevant findings that enhance understanding of CPP presentation according to age, thereby improving clinical management of this condition.

Chlorogenic acid improves urogenital dysfunction induced by exposure to ambient particulate matter.

Oxidative stress is a well-known underlying mechanism for several diseases in response to environmental pollution. Although there is a lack of evidence on the relationship between air pollution and an established risk factor for urogenital dysfunction. The aim of this study was to investigate the mechanism of particulate matter (PM) on urogenital function and evaluate the potential efficacy of chlorogenic acid (CGA) in preventing urogenital damage in rats. Forty Wistar rats were divided into five groups (n = 8): control, particulate matter exposure (animals were exposed to fine dust in an inhalation chamber for 4 weeks, 3 days a week, for 3 h, PM10 concentration adjusted to 500-2000 µg/m3), and particulate matter plus 3 concentrations of chlorogenic acid (100, 200, and 400 mg/kg, gavage, 4 weeks, 3 days a week). At the end of the study, kidney biomarkers, oxidative stress markers, antioxidant enzymes, the oxidation resistance 1 (OXR1) and its downstream gene expression, sperm count, gonadotropin hormones, and the structure of the kidney, epididymis, and seminal vesicle were evaluated in response to PM exposure and CGA treatment in all groups. The data obtained from the current study showed that PM exposure led to kidney dysfunction and inhibition of oligospermia through oxidative stress, as evidenced by an increase in MDA and a decrease in TAC, SOD, CAT, and GSH concentration levels in blood samples. These results were consistent with the down-regulation of OXR1, Nrf2, and P21 gene expression. In contrast, CGA improved urogenital biomarkers and histopathology structures of the kidney, epididymis, and seminal vesicle by enhancing antioxidant defense system enzymes and modulating the OXR1 signaling pathway. Our findings suggest that environmental air pollution contributes to kidney dysfunction and urogenital damage. Modulation of oxidative stress through the OXR1, P21, and Nrf2 signaling pathways may be the underlying mechanism. Furthermore, chlorogenic acid supplementation could be recommended as a new protective or treatment strategy to safeguard urogenital function against exposure to particulate matter.

Pregnancy detection based on blood serum sample Raman spectroscopy.

In this research, women were diagnosed as pregnant based on blood serum samples Raman spectroscopy. Raman spectroscopy is a vibrational technique that provides information on the chemical composition of samples. The Raman techniques have significantly impacted the study of various degenerative diseases, particularly cancer detection, using less invasive methods such as the analysis of blood serum samples. Additionally, these techniques have been used to study the health status of patients, which is often difficult to monitor using conventional techniques. This study obtained serum samples from 12 women diagnosed as pregnant and 11 non-pregnant volunteers (controls). Spectra were collected using a LabRAM HR800 Raman Spectrometer (Horiba Jobin-Yvon) with an 830 nm wavelength laser. For each serum sample, 10 Raman spectra were obtained by focusing the laser, using a microscope objective, on different points of the sample with an exposure time of 40 seconds and an irradiation power of 17 mW. The raw spectra were processed using baseline correction and smoothing to remove noise, fluorescence, and shot noise. Subsequently, the spectra were normalized and analyzed using the multivariate statistical method of Principal Component Analysis (PCA). In these spectra, the characteristic bands of main blood serum biomolecules such as phenylalanine (Phe), tyrosine (Tyr), glutathione, amide III, phospholipid, carotene, and tryptophan (Trp) can be observed. Nevertheless, when analyzing the average spectra of pregnant and non-pregnant women, the main spectral differences were associated with variations in molecules like glutathione, tryptophan (Trp), tyrosine, and phenylalanine, which occur during the first trimesters of pregnancy. This aligns with findings previously reported by other studies. Furthermore, the serum samples from pregnant and non-pregnant patients can be effectively discriminated using PCA applied to the Raman spectra, revealing two distinct clusters in the PCA plot corresponding to each group's status. The results demonstrate that pregnancy can be determined based on blood serum samples Raman spectroscopy with sensitivity and specificity. Although there are very effective devices on the market to determine pregnancy based on the Human Chorionic Gonadotropin (HCG) hormone detection in urine samples, these preliminary results indicate an alternative method known as Raman spectroscopy. On the other hand, the results could also suggest the possibility of carrying out other gynecological or fetal-related studies in women using these Raman techniques.

Identification and Characterization of Novel FSHR Copy Number Variations Causing Premature Ovarian Insufficiency.

Follicle stimulating hormone (FSH) is a key pituitary gonadotropic hormone implicated in human fertility and is crucial for folliculogenesis and recruitment of new antral follicles. Variations in its receptor, FSHR, can lead to diverse reproductive phenotypes including ovarian hyperstimulation syndrome (OHSS) and premature ovarian insufficiency (POI). This study reports a novel case of FSHR-related ovarian insufficiency in a patient with primary amenorrhea, subnormal AMH levels, and delayed puberty. Genetic exploration revealed two compound heterozygous intragenic deletions of FSHR. Specifically, the patient inherited a maternally derived deletion spanning exons 5-10 and a paternally derived deletion involving exons 3-6. Through chromosomal microarray analysis (CMA), exome sequencing, long-range PCR, and Sanger sequencing, we characterized the breakpoints and confirmed the compound heterozygous deletions. The findings reveal a complete loss of function of both FSHR alleles, contributing to the patient's POI phenotype. This case emphasizes the complexity of genotype-phenotype correlations in FSHR-related disorders and the role of CNVs in POI phenotypes. Although these events are rare, our results advocate for the inclusion of CNV detection in the diagnostic workup of POI to ensure accurate diagnosis and better patient management.

Histomorphometric changes in pituitary gonadotropic endocrinocytes when exposed to dark deprivation

Aim. To assess the effect of 30-day dark deprivation on functional and histomorphometric changes in adenohypophysis gonadotropic endocrinocytes and their reversibility in mature male rats.Materials and methods. Mongrel white male rats (n = 36) weighing 365–375 g at 4 months of age were randomly divided into three groups (each n = 12). For 30 days the control group was in automatic light-dark mode 12/12, and the rats of experimental groups 1 and 2 were in round-the-clock artificial lighting (24/0, 300 Lux), then the rats of group 2 were returned to 12/12 mode for the next 14 days. In the animals of the control and group 1 during their lifetime on the 31st day, and in group 2 on the 45th day, blood was taken from the abdominal aorta and levels of follicle-stimulating (FSH) and luteinizing (LH) hormones, melatonin, and Klotho protein were determined an enzyme-linked immunosorbent assay and immunoassay and after which they were removed from the experiment by decapitation. Postmortem histological and immunohistochemical examination of the pituitary gland was done using rabbit polyclonal antibodies targeting caspase-3 and Klotho protein, as well as morphometry. Statistical data processing was performed using the Kruskal-Wallis test with post-hoc Dunn’s test.Results. Light desynchronization in the form of 30 days of dark deprivation increased FSH and LH levels and decreased melatonin and Klotho protein levels in the blood of male rats; increased gonadotropic endocrine cell area, volume, and perimeter by 23.1% (p &lt; 0.001), 48.7% (p &lt; 0.001), and 10.9% (p &lt; 0.001), respectively; and increased nucleus area, volume, and perimeter by 16%, 11.7%, and 2.5%, respectively. An immunohistochemical study showed an increase in the specific area of caspase-3-immunoreactive gonadotropic endocrinocytes by 25.2% without obvious morphological signs of apoptosis, and a decrease in the expression of Klotho protein by 25.7%. All indicators were reversible, the levels of FSH and Klotho protein in the blood of animals almost reached their initial values after 14 days of restoration of the light-dark cycle 12/12.Conclusion. Dark deprivation for 30 days in male rats induced reversible processes of accelerated aging and apoptosis in cells, as evidenced by changes in the expression of aging markers in gonadotropic endocrinocytes and levels of gonadotropic hormones in the blood. When the light-dark mode is restored, the levels of FSH and Klotho protein normalize as early as 14 days.

Transcription factors NEUROD1 and NEUROD4 in the migration of gonadotrope cells during pituitary embryogenesis

Transcription factors NEUROD1 and NEUROD4 in the migration of gonadotrope cells during pituitary embryogenesis

An optimized fractionation method reveals insulin-induced membrane surface localization of GLUT1 to increase glycolysis in LβT2 cells

Insulin is an important regulator of whole-body glucose homeostasis. In insulin sensitive tissues such as muscle and adipose, insulin induces the translocation of glucose transporter 4 (GLUT4) to the cell membrane, thereby increasing glucose uptake. However, insulin also signals in tissues that are not generally associated with glucose homeostasis. In the human reproductive endocrine axis, hyperinsulinemia suppresses the secretion of gonadotropins from gonadotrope cells of the anterior pituitary, thereby linking insulin dysregulation to suboptimal reproductive health. In the mouse, gonadotropes express the insulin receptor which has the canonical signaling response of IRS, AKT, and mTOR activation. However, the functional outcomes of insulin action on gonadotropes are unclear. Here, we demonstrate through use of an optimized cell fractionation protocol that insulin stimulation of the LβT2 gonadotropic cell line results in the unexpected translocation of GLUT1 to the plasma membrane. Using our high purity fractionation protocol, we further demonstrate that though Akt signaling in response to insulin is intact, insulin-induced translocation of GLUT1 occurs independently of Akt activation in LβT2 cells.

The female reproductive tract microbiome and obesity

High-throughput 16S rRNA sequencing has allowed us to identify novel microorganisms and their relationships in the female reproductive tract. However, in obese patients, the female reproductive tract microbiome, unlike the intestinal microbiome, has been understudied. Here, the literature review analyzes and describes microbiome features in the external genitalia, vagina, cervical canal, uterus, and ovaries in overweight and obese nonpregnant and pregnant females. The microbiome of the lower female reproductive tract in obese patients is characterized by increased bacterial diversity, pH, decreased Lactobacillus abundance, and increased abundance of obligate anaerobes and yeasts of the genus Candida. The endometrial microbiome in overweight and obese patients has been studied only in postmenopause and is characterized by higher Proteobacteria abundance. No data on the characteristics of the ovarian microbiome in obese patients are available. The mechanisms accounting for microbiome changes in obese patients are likely to due to the ability of adipose tissue-derived leptin and estrone to inhibit production of pituitary gonadotropic hormones resulting in blocked ovulation and lowered estradiol production in patients of reproductive age. Consequently, a decline in glycogen synthesis in the vaginal epithelium, decreased Lactobacillus abundance followed by elevated vaginal mucus pH value and number of obligate anaerobes, including those associated with bacterial vaginosis are observed. Weight loss can have a beneficial effect on the state of the vaginal microbiome, restoring normal Lactobacillus abundance.

Gestational Trophoblastic Tumours: Experience of the Oncology Department of the Moulay Ismail Military Hospital in Meknes (10 Cases)

Gestational trophoblastic tumors (GTTs) are the malignant forms of gestational trophoblastic diseases. They always follow a pregnancy, most often molar. The diagnosis of GTT is based on an abnormal course of HCG (chorionic gonadotropin hormone) and/or signs of ultrasound invasion and/or histological confirmation. We report a retrospective series of 10 cases of GTT collected at the Medical Oncology Department of the Moulay Ismail Military Hospital in Meknes over an 8-year period from January 2012 to January 2020. The analysis of our series led to the conclusion that: The average age of onsetis 42 years. 90% of our patients were pauciparic. The causal pregnancy was in 90% of the cases a molar pregnancy. The extension assessment was mainly based on the TAP CT identifying the lung as the main metastatic site in 40% of cases. 5 patients (50% of cases) underwent hysterectomy with post operative chemotherapy. 60% of our patients were classified as low risk according to the FIGO score and received methotrexate mono-chemotherapy. 4 patients were classified as high-risk and were treated with poly chemotherapy, the main protocol of which was EMA-CO. All our patients benefited from clinical and biological monitoring, before each chemotherapy session; then weekly and monthly after negativation, up to 12 months in case of good prognosis GTT, and up to 18 months in case of bad prognosis GTT. One case of MTX relapse (the case of IPTT) was reported and for which a 2nd line CMT with a favourable evolution was indicated. This study allowed us to analyze the cure rate, which was 100% for patients who were treated and had completed their monitoring.

6873 Expression of Constitutively Active YAP in Murine Gonadotrope Cells Leads to Morphological Alterations and Dysfunction of the Anterior Pituitary Gland

Abstract Disclosure: N. Jakuc: None. M. Bérubé: None. E. Corrêa Dos Santos: None. G. St-Jean: None. M. Paquet: None. U. Boehm: None. D.J. Bernard: None. D.T. Boerboom: None. A. Boyer: None. G. Zamberlam: None. The Hippo transcriptional coactivators, YAP and TAZ, are expressed in cells throughout the anterior pituitary gland in mice. Although the potential physiological roles of YAP and TAZ in gonadotrope cells was unknown, we recently reported that conditional deletion of YAP and TAZ in gonadotrope cells results in increased circulating levels of LH (in males and females) and FSH (males only), and female hyperfertility in mice. Here, to further study the effects of Hippo signaling in gonadotropes, we conditionally expressed a constitutively active YAP (5SA) in these cells using the Cre-lox system with one of two Cre-driver strains, steroidogenic factor-1 (SF-1)-Cre or GnRH receptor-IRES-Cre (GRIC). Such crosses generated two distinct mouse models, which are referred to herein as YAP5SA;SF1-Cre and YAP5SA;GRIC, respectively. Expression of total YAP (YAP5SA and endogenous Yap) and the classic YAP-TEAD target genes, Ctgf, Cyr61, and Ankrd1 was significantly increased in pituitaries of 6-week-old mutant males and females in both the YAP5SA;SF1-Cre and YAP5SA;GRIC models. Animals from both models also showed significantly reduced serum LH and FSH levels as well as lower body mass and gonadosomatic indices when compared to respective controls. Six-week-old male and female mutants from both models showed significant morphological alterations in the pituitary. YAP5SA;SF1-Cre mutants exhibited proliferation of poorly differentiated cells, with high cellular and nuclear atypia with several mitoses. Similarly, YAP5SA;GRIC pituitaries were characterized by an increased number of poorly differentiated cells as well as follicle or papillary-like structure formation and increased cell proliferation and necrosis. Together, these preliminary findings strongly suggest pituitary blastoma or choriocarcinoma, and confirmatory experiments are underway. In light of gross morphological changes that appeared to extend beyond gonadotropes, we performed qPCR analysis of other cell lineages in 6-week-old YAP5SA;GRIC mutants and corresponding controls. Gh, Prl, Tshb, and Pomc mRNA levels were decreased in pituitaries of mutant animals. In addition, GH and ACTH protein expression, as assessed by immunohistochemistry, was nearly undetectable in pituitaries in mutant animals, suggesting compromised presence or function of somatotropes and corticotropes. To better understand the onset of this pathology, we performed histopathological analysis of YAP5SA;GRIC mutants in early post-natal life. Although pituitaries from 1-day-old mutant animals seemed normal in comparison to controls, we found signs of cellular necrosis and vacuolar degeneration in 1-week-old mutants. Together, our results indicate that constitutively active YAP in murine gonadotropes leads to significant morphological and functional alterations throughout the pituitary. Presentation: 6/1/2024

6629 Pure Androgen Secreting Adrenal Adenoma In A Premenopausal Woman

Abstract Disclosure: S. Syed: None. Background: Pure androgen secreting adrenal adenomas are extremely rare; most of the reported cases occur in post-menopausal women, with significant virilization of acute onset and are usually malignant. Here we present a case of an androgen secreting adrenal adenoma found in a young female with regular menstrual cycles and hirsutism. Clinical Case: A 27-year-old female with no significant past medical history was referred to Endocrinology clinic for evaluation of elevated testosterone level noted on a routine OBGYN exam. Patient had miscarriage six months prior to initial endocrine clinic visit. She had a routine follow up visit with her gynecologist after miscarriage and noted to have mild male pattern hair growth on her chin, worse after stopping birth control pills two years ago. Patient had menarche at 14 years of age and had regular menstrual cycle until started birth control pills. The skin examination revealed coarse male pattern hair on chin. Scattered acne was present on back. No signs of hypercortisolism were present. Labs done at time of initial consultation showed unremarkable comprehensive metabolic panel, thyroid and gonadotropic hormones. Testosterone was elevated at 122ng/dl (0-75ng/dl). Pelvic ultrasound done in the year prior showed normal uterus and adnexal structures. Further work up showed elevated DHEAS 781 µg/dL (18-391 µg/dL) and normal 17 OH progesterone and androstenedione levels. MRI abdomen with and without contrast done at outside facility showed a 4 cm x 3.3 cm x 4.1 cm well-circumscribed round shaped, mildly heterogeneously enhancing mass arising from right adrenal gland, with restricting diffusion, no signal dropout in out-of-phase imaging. Normal ovaries were noted. Given the large size of lesion, adrenalectomy was recommended. Patient underwent laparoscopic right adrenalectomy. The histologic features on pathology specimen were consistent with an adrenal cortical adenoma. No evidence of malignancy was seen (based on modified Weiss criteria). 3 weeks post op labs showed Testosterone level reduced to 19 ng/dl (0-75ng/dl) and DHEAS 138 ug/dl (18-391 µg/dL). Patient is recovering from recent childbirth. Conclusion: In adults, pure androgen secreting adrenal tumors are extremely rare. Diagnosis of malignancy is made with histopathological examination and Weiss score calculation. Our patient has unique presentation of having mild hirsutism in setting of regular menstrual cycles with no other symptoms of virilization and is noted to have unexpectedly large adrenal adenoma. Given the size of adenoma, there was concern for adrenal cortical carcinoma, however histological examination was consistent with a benign lesion. Presentation: 6/1/2024

8574 Single cell multiomics suggests tumor cell energy metabolism heterogeneity in non-functioning gonadotroph pituitary adenomas

Abstract Disclosure: F.M. Ruf-Zamojski: None. Z. Zhang: None. W. Cheng: None. G.R. Smith: None. M. Zamojski: None. N. Mendelev: None. Y. Ge: None. D. Marrero-Rodríguez: None. K. Taniguchi-Ponciano: None. C.L. Andoniadou: None. E. Zaslavsky: None. X. Chen: None. O.G. Troyanskaya: None. M. Mercado: None. S.C. Sealfon: None. Pituitary adenomas cause hormonal dysregulation and severe morbidity. The absence of clear biomarkers for prognosis/treatment and the high risk of recurrence make management challenging. Non-functioning gonadotroph pituitary adenomas (NFPAs) represent proliferation of gonadotroph lineage cells without an increase in secretion of gonadotropins. Detection is often delayed until mass effects cause visual defects. Higher resolution molecular study may improve classification, diagnosis, treatment, and provide insight into adenoma biology. We analyzed four NFPAs using unbiased genome-wide same-cell single nucleus 10X Genomics multiome assay (transcriptome and chromatin accessibility) to define individual cell identity/states in comparison with normal gonadotrope cell references from twelve age/sex-matched human post-mortem pituitaries. In total, we generated high quality multiomics single-cell data for 33,832 adenoma cells and 5,451 normal gonadotropes. We identified tumor, immune, vascular, and proliferative cells confirming cellular heterogeneity. Multidimensional scaling analysis using Manhattan distances between cell types showed that NFPA gonadotropes were closer in gene expression to healthy gonadotropes among all pituitary cell types. In addition, we observed differences in the macrophage and endothelial cell populations between tumors and healthy pituitaries. Next, we studied gene regulatory circuitry (regulated gene, modulating transcription factor and its chromatin interaction site) using the new Control of Regulation Extracted from Multiomics Assays (CREMA, [1]) multiome analysis method which showed high variation between normal and tumor gonadotropes. Within each adenoma, the NFPA tumor cells encompassed two main subgroups of cells, one group (GT+ marker tumor cells) expressing established gonadotrope markers and another group not (GT- marker tumor cells). GT+ and GT- tumor cells formed a cluster distinct from healthy gonadotropes. GT+ and GT- tumor cells differed by their level of expression of mitochondrial genes, suggesting decreased aerobic capacity associated with the GT- tumor subgroup. Interestingly, differential analyses between the healthy and the NFPA gonadotropes from same-cell multiome data highlighted dissimilar gene expression and chromatin accessibility patterns.Our newly collected single cell data characterize the difference between NFPA tumor cells and normal gonadotrope cells. Our results suggest a Warburg-effect like difference in aerobic respiratory capacity among tumor cells within the same pituitary adenoma. Overall, this study brings new insights into the molecular characteristics of NFPAs, including tumor cell heterogeneity, and represents a new resource for the field. [1] PMID: 38084892. Presentation: 6/1/2024

Comparison of highly purified human menopausal gonadotropin and recombinant follicle stimulating hormone use in patients undergoing in vitro fertilization with progestin-primed ovarian stimulation protocol: a single center retrospective analysis.

Recently, progesterone has been used to prevent LH surge instead of GnRH analogues during ART treatments, which is known as progesterone-primed ovarian stimulation (PPOS) protocol. During ART treatment, highly purified human menopausal gonadotropin (HP-hMG) and recombinant follicle stimulating hormone (rFSH) are two of the agents used for stimulation of antral follicles. The aim of this study is to compare the efficacy and success of HP-hMG and rFSH agents in the ovarian stimulation step of the PPOS protocol, which has not been previously reported in the literature. This retrospective study was conducted at a university hospital with patients who underwent IVF treatment using PPOS protocols in between January 2019 and July 2021. For ovarian stimulation, rFSH was used in group I and HP-hMG was used in group II. Mature oocyte ratio was the primary outcome, and live birth rate was the secondary outcome. Mann-Whitney and Chi-square tests were used for statistical analysis. All p values below 0.05 were considered significant. Total numbers of follicles, oocytes, MII, and 2PN numbers obtained were similar between the two groups. The fertilization rates were 66.7% in the rFSH group and 64.3% in the HP-hMG group (p > 0.05). The pregnancy rates were 53.5% and 46.7% in the rFSH and HP-hMG groups, respectively. There was no statistically significant difference between pregnancy, abortus, and live birth rates. In this study, it is demonstrated that stimulation of oocytes with either rFSH or hMG in the PPOS protocol, which has been added to IVF treatment protocols in recent years, had no statistical difference regarding mature oocyte numbers and live birth rates between the two groups. These results are consistent with the previous literature which compared rFSH and hMG in GnRH agonist and antagonist protocols.

The impact of chronic stress on the hormonal state of women affected by hostilities and displaced women

During times of war, refugee women often have high levels of chronic stress due to factors such as war, displacement and uncertainty. The study focuses on examining the hormonal responses of female refugees experiencing chronic stress from war and displacement.The objective: to assess the impact of chronic stress caused by war and displacement on the hormonal balance of refugee women.Materials and methods. 60 women were examined, who were divided into groups: the 1st group included women who were affected by hostilities (20 persons), and the 2nd group included displaced women (20 individuals). In these 40 women the hormonal studies were conducted in the conditions of military aggression in Ukraine. The control group (3rd group) included healthy women who were examined in the pre-war period. All examined women were 20–25 years old with an average body weight of 55–60 kg. Concentrations of progesterone, estradiol, follicle-stimulating hormone (FSH) and luteinizing hormone, prolactin in the I and II phases of the menstrual cycle (MC), cortisol in the I phase of the MC were determined in blood serum using the enzyme immunoassay method. An analysis of the results of research on the role of the hypothalamic-pituitary-adrenal system in the regulation of the stress response was carried out.Results. In the examined women, various disorders of MC were observed, such as: menometrorrhagia, hypermenorrhea, acyclic uterine bleeding, oligomenorrhea, and amenorrhea. The most pronounced changes in menstrual function were observed in women who were affected by hostilities. Such significant disorders of MC in women of this group coincided with changes in indicators of gonadotropic hormones of the pituitary gland and sex hormones in the blood.A study of the psycho-emotional state of women who suffered from hostilities established that these women have a significant stressful effect of various risk factors on the body. Changes in the levels of cortisol, prolactin and other hormones were detected.Conclusions. Studies have shown that chronic stress affects the hormonal status of women affected by hostilities and of displaced women, including FSH, estradiol, and prolactin levels.A significant increase in the level of FSH in the follicular phase of the menstrual cycle and an increase in the level of estradiol in the luteal phase were determined, which may indicate a compensatory reaction of the body to stress. A significant increase in the level of prolactin was also found, which can cause disorders of menstrual function and affect the psycho-emotional state.

Investigation of the Effect of Nano-melatonin on Oxidative Stress in Sera of Patients with Polycystic Syndrome

Background: The association between polycystic ovary syndrome (PCOS) and oxidative stress in women, the possible causes of PCOS, insulin resistance (IR), obesity, and oxidative stress. Oxidants are chemical elements that tend to gain electrons, losing positive charge. The Oxidative Stress (OS) is defined as an imbalance between the formation and synthesis of free radicals and the antioxidant defense system, which results in oxidative damage. Objective: The purpose of this research is to investigate the effect of nano-melatonin on oxidative stress in the sera of patients with polycystic syndrome. Methods: The study includes 120 Iraqi women, divided into two groups: 60 PCOS patients and 60 healthy control patients. Sample Analysis Biochemistry analyses the hormonal profile and determines the hormonal levels (LH, FSH, Testosterone, and Prolactin) that were identified for all controls and patients. Hormones analysis was performed using the enzyme-linked fluorescent assay (ELFA) technique. Synthesized ML nanoparticles by ultrasonic sonication. Results: The results indicate that the mean of serum LH significantly increased (P&lt;0.02) in PCOS BMI ≥ 25 patients G1 (6.19±3.13) mIU/mL compared with controls C1 (4.504±1.591) mIU/mL, while a nonsignificant difference (P&gt;0.05) when compared the BMI&lt;25 kg/m2 for G2 (5.55±1.93) with control group C2 (6.31±2.58) of luteinizing hormone in G1 and G2. Conclusions: An increased ratio of LH to FSH was shown to be characteristic of PCOS. For PCOS, high LH values are crucial. For the purpose of diagnosing PCOS, high LH values are crucial release of gonadotropin hormones (LH, FSH) from the pituitary. To higher the production of androgen in ovarian theca cells, The LH receptor is activated by the luteinizing hormone (LH). The nano Mel was higher in the inhibition of Oxidative stress two causes nano-melatonin high surface area and small size.

Characterization of gonadotropins and their receptors in a chondrichthyan, Scyliorhinus canicula, fills a gap in the understanding of their coevolution

In Gnathostomes, reproduction is mainly controlled by the hypothalamic-pituitary–gonadal (HPG) axis, with the involvement of the pituitary gonadotropic hormones (GTH), follicle-stimulating hormone (FSH) and luteinizing hormone (LH), which activate their cognate receptors, FSHR and LHR, expressed in gonads. Each GTH consists of a common α subunit and of a specific FSHβ or LHβ subunit. Chondrichthyes (holocephalans and elasmobranchs) is a sister group of bony vertebrates. This position is highly favorable for the understanding of the evolution of endocrine regulations of reproduction among gnathostomes. Surprisingly, the characterization of gonadotropins and their receptors is still limited in chondrichthyes. In the present study, GTH and GTHR sequences have been identified from several chondrichthyan genomes, and their primary structures were analyzed relative to human orthologs. 3D models of GTH/GTHR interaction were built, highlighting the importance of the receptor hinge region for ligand recognition. Functional hormone-receptor interactions have been studied in HEK cells using the small-spotted catshark (Scyliorhinus canicula) recombinant proteins and showed that LHR was specifically activated by LH whereas FSHR was activated by both FSH and LH. Expression profiles of GTHs and their receptors were explored by real-time PCR, in situ hybridization and immunohistochemistry during spermatogenesis, along the male genital tract and other tissues, as well as in some female tissues for comparison. Tissue-expression analyses showed that the highest levels were observed for fshr transcripts in testis and ovary and for lhr in specific extragonadal tissues. The two receptors were expressed at all stages of spermatogenesis by both germ cells and somatic cells, including undifferentiated spermatogonia, spermatocytes, spermatids, somatic precursors and Sertoli cells; differentiated Leydig cells being absent in the testis of S. canicula. Receptors were also expressed by the lymphomyeloid epigonal tissue and the testicular tubules. These results, suggest a wide range of gonadotropin-regulated functions in Elasmobranchs, as well as functional redundancy during spermatogenesis. These extended functions are discussed in an evolutionary context in which the specificity of gonadotropin signaling must have contributed to the evolution of gonadal cells’ morphology and function.