Gonadotroph Cell Research Articles (Page 1)

Neuroendocrine pituitary tumors (PitNETs) are classified based on clinical manifestation and expression of pituitary cell lineage-specific transcription factors (TFs) and hormones. A subtype of tumors in patients with acromegaly was found to express PIT-1 and SF-1 TFs, two markers of distinct pituitary cell lineages. These tumors have been described as multilineage or “somatogonadotoph” tumors. The aim of our study was to clarify their identity and cell type origin using extensive transcriptomic analysis. For this purpose, we analyzed the RNA sequencing (RNAseq) data from 546 PitNETs (including 193 tumors of patient with acromegaly) and single cell RNAseq data from somatotroph and gonadotroph tumors and normal pituitary tissue. Somatrotroph PitNETs co-expressing PIT-1 and SF-1 TFs were identified in each of the analyzed RNAseq dataset. Their transcriptomic profile and pituitary TF activity closely resembled those of other somatotroph tumors, while differing substantially from gonadotroph PitNETs, though they retained NR5A1 (SF-1) activity and expressed some SF-1-regulated genes (e.g., LHB and GNHRH). Notably, SF-1 appeared to regulate a slightly different set of genes in double positive somatotroph PitNETs and gonadotroph tumors. Analysis of scRNAseq data revealed a subcluster of normal gonadotroph cells expressing POU1F1 (PIT-1), but tumor cells from PIT-1/SF-1 PitNETs did not resemble this normal gonadotroph cell subtype. Genes expression profiles of three subtypes of somatotroph tumors were distinguished through analyses of both bulk- and scRNAseq data. From transcriptomic perspective — based on gene co-regulation and pituitary TF activity — PitNETs of patients with acromegaly that co-express PIT-1 and SF-1 represent a subtype of PIT-1 lineage tumors, and the molecular data do not support classifying them as multilineage tumors.Supplementary InformationThe online version contains supplementary material available at 10.1186/s40478-025-02091-z.

BACKGROUND: The current clinical classification and treatment principles for pituitary adenomas are primarily based on the analysis of hormone levels in blood plasma. However, this approach does not account for the heterogeneity and plurihormonal nature of these tumors, which pose significant diagnostic challenges. Immunohistochemical (IHC) analysis is a crucial tool for studying the molecular characteristics of resected tumors, enabling the identification of features that may not be apparent through standard biochemical tests and clinical evaluation.AIM: To investigate the heterogeneity and hormone co-expression of pituitary adenomas through a comparative analysis of clinical, hormonal, and immunohistochemical data, aiming to optimize the diagnosis and classification of pituitary neuroendocrine tumors.MATERIALS AND METHODS: The study included 10 pituitary adenoma samples. A comprehensive immunohistochemical analysis was performed to evaluate the expression of transcription factors and pituitary hormones to detect tumor heterogeneity and potential plurihormonal pituitary adenomas.RESULTS: The analysis confirmed significant heterogeneity of pituitary adenomas and identified cases of hormone co-­expression in certain tumors, which may be overlooked when relying solely on clinical data and blood test results, or in the absence of IHC testing that covers all hormones and transcription factors in pituitary tissue. In our study, we identified heterogeneity in pituitary adenoma samples at the level of transcription factor and hormone expression. For example, in two hormonally active corticotropinomas, we observed not only the expression of standard IHC markers but also the unexpected expression of GH and PIT1. Notably, somatotropinomas exhibited distinct expression of SF1, a transcription factor typically specific to gonadotroph cells. Additionally, we identified signs of heterogeneity in hormone-inactive pituitary tumors — such as silent gonadotropinomas — where co-expression of both SF1 and TPIT was detected.CONCLUSION: The results of the study confirmed significant heterogeneity of pituitary adenomas and the presence of hormone co-expression, indicating the complexity of their diagnosis when relying solely on clinical and hormonal methods. Comprehensive immunohistochemical analysis plays a key role in the accurate classification of these tumors, which may contribute to the improvement of diagnostic and therapeutic approaches.

INTRODUCTION: The cellular origins of pituitary neuroendocrine tumors (PitNETs) are uncertain, positing that PitNETs may originate from differentiated neuroendocrine cells or adult pituitary stem cells. Understanding these origins is essential for developing targeted therapies aimed at tumor-initiating cells, thereby enhancing treatment efficacy. METHODS: We conducted single-nucleus sequencing on approximately half a million cells, creating the largest database to date. This included samples from two non-tumor postmortem adult pituitaries and 11 resected PitNETs, covering somatotrophic, hormonally active and silent corticotrophic (SCAs), lactotrophic, silent gonadotroph (SGA), and null cell (NCA) adenomas. RESULTS: We reconstructed a single-nuclei atlas of the normal adult human pituitary gland, identifying pituitary stem cells expressing previously known and novel transcription factors. Integrating stem and neuroendocrine nuclei (corticotroph, gonadotroph, lactotroph, thyrotroph, and somatotroph) from normal controls with tumor samples traced the development of PitNET subtypes, with copy number variation (CNV) analysis used to assess chromosomal instability. While most PitNET subtypes derived from their differentiated normal counterpart cells (SGAs from gonadotroph cells, prolactinomas from lactotroph cells, SCA/secreting corticotrophic adenomas from corticotroph cells) and had significant CNV changes, somatotroph and null cell adenomas had minimal CNV changes and arose from pituitary stem cells. While secreting corticotrophic adenomas and SCAs originated from mature corticotroph cells, secreting corticotrophic adenomas appeared earlier in developmental trajectory, representing a less mature state than SCAs. On gene expression analysis, secreting corticotrophic tumors upregulated genes associated with cell morphogenesis and development, suggesting a non-terminal transitional state. Lastly, we characterized key pseudotime genes for each tumor subtype. CONCLUSIONS: This study provides detailed insight into the cellular origins and developmental pathways of various PitNET subtypes, revealing that both stem and differentiated cells can contribute to tumorigenesis in a subtype-specific manner.

Exposure to environmentally relevant levels of DEHP during development modifies the distribution and expression patterns of androgen receptors in the anterior pituitary in a sex-specific manner.

Gonadotroph neuroendocrine pituitary tumors are among the most common intracranial neoplasms. A notable proportion of these tumors is characterized by invasive growth which hampers the treatment results and worsens prognoses of patients. Increased hsa-miR-184 expression was observed in invasive as compared to non-invasive gonadotroph tumors. This study aimed to determine the role of hsa-miR-184 expression in invasive growth of gonadotroph tumors. QRT-PCR and bisulfite pyrosequencing were used for evaluating hsa-miR-184 expression and MIR184 DNA methylation levels, respectively, in tumorsand normal pituitary samples. LβT2 and αT3-1 gonadotroph cells were used to test the effect of miR-184 on cell viability (MTT test), proliferation (BrdU incorporation), and migration (scratch assay). RNA sequencing was applied for transcriptome profiling in miR-184-treated and untreated LβT2 cells. Differential genes expression analysis combined with target prediction served for identification ofmiR-184 targets. MiRNA-mRNA interaction was subsequently validated with Luciferase reporter assay. Analysis of tissue samples showed that hsa-miR-184 is upregulated in gonadotroph tumors and its expression is higher in invasive than in noninvasive ones. Promoter of MIR184 is demethylated intumors, and the methylation level is negatively correlated with hsa-miR-184 expression. Transfecting LβT2 and αT3-1 with miR-184 mimic resulted in increased cellular proliferation and viability. Differentially expressed genes were identified when comparing miR-184-treated and untreated cells, including Nus1 as the only predicted miR-184 target. The interaction between miR-184 and 3'UTR of Nus1 was confirmed in vitro in both LβT2 and αT3-1. Overexpression of Nus1 resulted in lowering cell viability in both cell lines and proliferation in LβT2. The expression level of NUS1 was lower in invasive than in noninvasive tumors. Our results indicate that DNA hypomethylation-related increase of hsa-mir-184 expression contributes to invasive growth of gonadotroph pituitary tumors through targeting NUS1, being one of the various molecular mechanisms involved in conferring aggressive growth potential.

Abstract Disclosure: M. Muhammed: None. S. Freeman: None. M. Mahamdeh: None. M. Martinez-Lage Alvarez: None. K.K. Miller: Other; Self; Dr. Miller has received study medication and investigator-initiated research grants from Amgen and has equity in Bristol-Myers Squibb (BMS), General Electric, Boston Scientific, and Becton Dickinson.. R.J. Soberman: None. E.A. Lawson: Other; Self; Funding and study drug (intranasal oxytocin/placebo) from Tonix Pharmaceuticals;, U.S. provisional patent application no. 63/467,980 Oxytocin-Based Therapeutics to Improve Cognitive Control in Individuals with Attention Deficit Hyperactivity Disorder. Objectives: Oxytocin (OXT) is a hypothalamic-posterior pituitary hormone that induces pleotropic effects by binding to a G-protein-coupled receptor (OXTR) that is present across multiple organ systems. There is evidence that signaling by OXTRs modulates anterior pituitary function, increasing the release of prolactin, growth hormone, and gonadotroph hormone, while reducing the release of thyroid stimulating hormone and adrenocorticotropin hormone. However, it is not known whether OXTRs are present in the human pituitary gland. Studying OXTR distribution is challenging due to homology and cross reactivity between OXTRs and vasopressin receptors, and reliable OXTR-specific antibodies are not available. RNAscope technology is a reliable and validated in situ hybridization method that can detect gene expression with high specificity. Therefore, we aimed to determine whether OXTRs are present in the human anterior pituitary gland using OXTR-specific probes that are complimentary to OXTR mRNA and do not cross react with mRNA for vasopressin receptors. We hypothesized that OXTR mRNA would be co-expressed in cells also expressing prolactin, growth hormone, thyroid-stimulating hormone, pro-opiomelanocortin, or follicle stimulating hormone subunit genes, consistent with OXTR presence across anterior pituitary cell types. Methods: De-identified nonclinical human pituitary tissue was used. Pituitary tissue was obtained at autopsy (postmortem). Formalin-fixed paraffin embedded tissue blocks were sectioned at 5 μm and mounted on microscope slides. RNA in situhybridization experiments were performed using RNAscope Multiplex Fluorescent v2 assays (Advanced Cell Diagnostics, Newark, CA) according to the manufacturer’s instructions. The samples were stained with Opal570 (OXTR) and Opal690 (others) and imaged using a Yokogawa spinning disc confocal on a Nikon Ti2 inverted microscope, and acquired images were evaluated for co-expression using Fiji, an open source image processing software. Results: Using RNAscope in situ hybridization, we confirmed that anterior pituitary cell populations that expressed prolactin, growth hormone, thyroid-stimulating hormone, pro-opiomelanocortin, or follicle stimulating hormone subunit also co-expressed OXTR mRNA. Conclusions: These data support our hypothesis that OXTR is present on human anterior pituitary lactotroph, somatotroph, thyrotroph, corticotroph, and gonadotroph cells. Our results provide a rationale to elucidate the function and clinical relevance of OXTRs in the anterior pituitary gland. Presentation: 6/3/2024

We examined how the sex steroids influence the synthesis of gonadotropins. The effects of sex steroids estradiol (E2), progesterone (P4), and dihydrotestosterone (DHT) in pituitary gonadotroph cell model (LβT2 cells) in vitro and ovary-intact rats in vivo were examined. The effects of sex steroids on Kiss1 gene expression in the hypothalamus were also examined in ovary-intact rats. In LβT2 cells, E2 increased common glycoprotein alpha (Cga) and luteinizing hormone beta (Lhb) subunit promoter activity as well as their mRNA expression. Although gonadotropin subunit promoter activity was not modulated by P4, Cga and Lhb mRNA expression was increased by P4. DHT inhibited Cga and Lhb mRNA expression with a concomitant decrease in their promoter activity. During the 2-week administration of exogenous E2 to ovary-intact rats, the estrous cycle determined by vaginal smears was disrupted. P4 or DHT administration completely eliminated the estrous cycle. Protein expression of all three gonadotropin subunits within the pituitary gland was inhibited by E2 or P4 treatment in vivo; however, DHT reduced Cga expression but did not modulate Lhb or follicle-stimulating hormone beta subunit expression. E2 administration significantly repressed Kiss1 mRNA expression in a posterior hypothalamic region that included the arcuate nucleus. P4 and DHT did not modulate Kiss1 mRNA expression in this region. In contrast, P4 administration significantly inhibited Kiss1 mRNA expression in the anterior region of the hypothalamus that included the anteroventral periventricular nucleus. The expression of gonadotropin-releasing hormone (Gnrh) mRNA in the anterior hypothalamic region, where the preoptic area is located, appeared to be decreased by treatment with E2 and P4. Our findings suggest that sex steroids have different effects in the hypothalamus and pituitary gland.

Abstract Disclosure: J.N. Shapiro: None. T.X. Lu: None. J. Eapen: None. J.G. Karam: None. Gonadotroph adenomas are the most common subtype of nonfunctional pituitary adenomas, with 35 percent secreting enough LH or FSH to raise serum gonadotropin levels, but clinical syndromes due to hypersecretion of gonadotropins are rare. A low testosterone level with high FSH or LH can mislead to a diagnosis of concomitant primary hypogonadism rather than a secreting pituitary adenoma. We present a case of a patient with an FSH-secreting pituitary macroadenoma presenting with low testosterone levels. The patient is a 67-year-old man incidentally found to have a pituitary macroadenoma measuring 12 x 13 x 12 millimeters, impinging upon the optic chiasm on MRI of the brain obtained for headaches. Visual field tests revealed abnormalities compatible with glaucoma, but not chiasm compression. He has no symptoms of hormonal excess or deficit, although his wife reported decreased erectile function. Physical examination was unremarkable except for an atrophic left testicle. Pituitary hormone function tests (ACTH, AM cortisol, IGF-1, prolactin, thyroid function tests, and 24-hour urine free cortisol) were within normal range, except for an elevated FSH 14.8-18.1 mIU/mL and decreased testosterone in range of 185-256 ng/dL, with normal LH at 7.0 mIU/mL, for which he was referred for urology for evaluation of primary hypogonadism. The patient underwent transsphenoidal resection of the pituitary adenoma. Postoperative pathology revealed a pituitary adenoma, gonadotroph cell type, with immunohistochemical staining scattered positive FSH, and rare positive LH. Postoperative MRI showed post-surgical changes with no residual nodular enhancement. Following surgery, the patient suffered from severe clinical hypogonadism with episodes of profuse diaphoresis and extreme fatigue. Postoperative laboratory values revealed panhypopituitarism with undetectable testosterone level, and low LH and FSH levels. The patient was successfully treated with hydrocortisone, levothyroxine, and testosterone, with resolution of his symptoms. Our patient presented with a pituitary macroadenoma, low testosterone, and high FSH suggestive initially of primary hypogonadism, but found ultimately to have an FSH-secreting adenoma. He likely had underlying hypogonadism, primary or secondary, and although his adenoma secreted FSH, it was not enough to maintain testosterone level in normal range, possibly in part due to biologically inactive FSH molecules or to primary testicular failure. The extremely low testosterone levels postoperatively suggest that his testicular function was mostly stimulated by the adenoma secreted FSH and was therefore drastically lost after surgery. This case increases awareness to consider a gonadotropin-secreting tumor in the differential diagnosis of a pituitary adenoma with high FSH or LH, even when testosterone level is unexpectedly low. Presentation: Thursday, June 15, 2023

PurposeTransforming growth factor-beta receptor 3-like (TGFBR3L) is a pituitary enriched membrane protein selectively detected in gonadotroph cells. TGFBR3L is named after transforming growth factor-beta receptor 3 (TGFBR3), an inhibin A co-receptor in mice, due to sequence identity to the C-terminal region. We aimed to characterize TGFBR3L detection in a well-characterized, prospectively collected cohort of non-functioning pituitary neuroendocrine tumours (NF-PitNETs) and correlate it to clinical data.Methods144 patients operated for clinically NF-PitNETs were included. Clinical, radiological and biochemical data were recorded. Immunohistochemical (IHC) staining for FSHβ and LHβ was scored using the immunoreactive score (IRS), TGFBR3L and TGFBR3 were scored by the percentage of positive stained cells.ResultsTGFBR3L staining was selectively present in 52% of gonadotroph tumours. TGFBR3L was associated to IRS of LHβ (median 2 [IQR 0–3] in TGFBR3L negative and median 6 [IQR 3–9] in TGFBR3L positive tumours, p < 0.001), but not to the IRS of FSHβ (p = 0.32). The presence of TGFBR3L was negatively associated with plasma gonadotropin concentrations in males (P-FSH median 5.5 IU/L [IQR 2.9–9.6] and median 3.0 [IQR 1.8–5.6] in TGFBR3L negative and positive tumours respectively, p = 0.008) and P-LH (median 2.8 IU/L [IQR 1.9–3.7] and median 1.8 [IQR 1.1-3.0] in TGFBR3L negative and positive tumours respectively, p = 0.03). TGFBR3 stained positive in 22% (n = 25) of gonadotroph tumours with no correlation to TGFBR3L.ConclusionTGFBR3L was selectively detected in half (52%) of gonadotroph NF-PitNETs. The association to LHβ staining and plasma gonadotropins suggests that TGFBR3L may be involved in hormone production in gonadotroph NF-PitNETs.

Ovariectomy (OVX) causes a depletion of circulating estradiol (E2) and influences hypothalamic kisspeptin neurons, which govern gonadotropin-releasing hormone (GnRH) release and ultimately gonadotropin secretion. In this study, we examined the changes induced by OVX on the anterior pituitary gland in female rats. OVX significantly increased the mRNA expression of gonadotropin α, luteinizing hormone (LH) β, and follicle-stimulating hormone (FSH) β subunits within the pituitary gland compared with control (sham-operated) rats, and this was completely suppressed by E2 supplementation. High-dose dihydrotestosterone supplementation also prevented the OVX-induced increase in the expression of the three gonadotropin subunits. GnRH receptor mRNA expression within the pituitary was significantly increased in OVX rats, and this increase was completely inhibited by E2 supplementation. The mRNA expression of the receptors for adenylate cyclase-activating polypeptide and kisspeptin was unchanged by OVX. Although the mRNA levels of inhibin α, βA, and βB subunits within the pituitary gland were not modulated by OVX, follistatin gene expression within the pituitary gland was increased by OVX, and this increase was completely inhibited by E2 supplementation after OVX. In experiments using a pituitary gonadotroph cell model (LβT2 cells), follistatin itself did not modulate the mRNA expression of gonadotropin LHβ and FSHβ subunits, and the GnRH-induced increase in the expression of these genes was slightly inhibited in the presence of follistatin. Our current observations suggest that OVX induces several characteristic changes in the pituitary gland of rats.

Pituitary tumours are benign neoplasms that derive from hormone-producing cells of the pituitary gland. While medical treatments have emerged for most subtypes, Gonadotroph tumours that express FSH, and/or LH still lack therapeutic options apart from surgery and radiotherapy. Activin-ligands are physiological regulators of production and secretion of FSH by gonadotroph cells, but their role in gonadotroph tumorigenesis remains little explored. Using the LT2 mouse gonadotroph-cell line which produces FSH under Activin-stimulation, we first tested whether sub-cutaneous xenografts of LT2 cells resulted in tumour formation in Rag2KO mice. Histological analysis confirmed the presence of LT2-tumours with endothelial cells and macrophages in their microenvironment. FSH expression was found in a subset of clusters of LT2 cells in the tumours. We subsequently addressed the consequences of targeting activin-signalling via injection of a soluble activin decoy receptor (sActRIIB-Fc). sActRIIB-Fc treatment resulted in significantly decreased LT2 tumour volume. Reduced Smad2-phosphorylation as well as inhibition of tumour-induced FSH production confirmed the efficient targeting of activin-downstream signalling in treated tumours. More interestingly, treated tumours showed significantly fewer endothelial cells, associated with reduced Vegfa expression. In-vitro treatment of LT2 cells with sActRIIB-Fc had no effect on cell proliferation or apoptosis, but Vegfa expression was inhibited, pointing to a likely paracrine effect of LT2 cells on endothelial cells through activin-mediated Vegfa regulation. Further in-vitro and in-vivo studies are now needed to pinpoint the exact roles of activin-signalling in these processes prior to translating these observations to the clinic.

Pituitary adenomas (PAs) are the second most common primary brain tumor and may develop from any of the cell lineages responsible for producing the different pituitary hormones. DNA methylation is one of the essential epigenetic mechanisms in cancers, including PAs. In this study, we measured the expression profile and promoter methylation status of carbohydrate sulfotransferase 7 (CHST7) in patients with PA; then, we investigated the effect of the CHST7 methylation status on the proliferation and differentiation of PAs. The volcano map and Metascape results showed that the levels of CHST7 were related to the lineages’ differentiation and the cell adhesion of PAs, and patients with low CHST7 had greater chances of having an SF-1 lineage (p = 0.002) and optic chiasm compression (p = 0.007). Reactome pathway analysis revealed that most of the DEGs involved in the regulation of TP53 regulated the transcription of cell cycle genes (HSA-6791312 and HSA6804116) in patients with high CHST7. Correlation analysis showed that CHST7 was significantly correlated with the eIF2/ATF4 pathway and mitochondrion-related genes. The AUC of ROC showed that CHST7 (0.288; 95% CI: 0.187–0.388) was superior to SF-1 (0.555; 95% CI: 0.440–0.671) and inferior to FSHB (0.804; 95% CI: 0.704–0.903) in forecasting the SF-1 lineage (p < 0.001). The SF-1 lineage showed a higher methylation frequency for CHST7 than the Pit-1 and TBX19 lineages (p = 0.009). Furthermore, as the key molecule of the hypothalamic–pituitary–gonadal axis, inhibin βE (INHBE) was positively correlated with the levels of CHST7 (r = 0.685, p < 0.001). In summary, CHST7 is a novel pituitary gland specific protein in SF-1 lineage adenomas with a potential role in gonadotroph cell proliferation and lineage differentiation in PAs.

Abstract Study question There are no published series of OHSS due to FGA. What FGA features should clinicians look for during OHSS, and what treatments are effective? Summary answer FGA tumour size is always &amp;gt;10mm. Other pituitary hormones may be deficient. Surgical resection of FGA is an effective treatment for OHSS. What is known already Pituitary adenomas affect 1:1000 adults and are classified as functioning or non-functioning. Non-functioning pituitary adenomas do not secrete hormones, but most commonly stain histologically gonadotroph cells. Functional pituitary adenomas secrete hormones such as prolactin causing prolactinoma. However, it is rare for a pituitary tumour to cause clinical features of excessive gonadotrophins (functioning gonadotroph adenoma; FGA). Single case reports, but no case series, have been published on the presentation of FGA-induced OHSS in women. Surgical excision of adenomas has been reported to cause remission of symptoms, though systematic data are lacking owing to rarity of these tumours. Study design, size, duration National case series from tertiary neuroendocrine units in England, Wales and Scotland. Participants/materials, setting, methods Eight high-volume pituitary endocrine tertiary units within England, Wales and Scotland audited their records for any cases of FGA-induced OHSS; only seven patients have been identified to date. In all cases, there had been no recent exposure to assisted reproductive technologies (ART) or drugs known to induce OHSS including gonadotrophins or selective oestrogen receptor modulators (SERMS). Main results and the role of chance Seven cases of FGA were identified with mean age 31.6 years (range 16-48) at diagnosis. Two-of-seven women presented acutely unwell with abdominal pain, distention and palpable mass requiring oophorectomy for ovarian torsion/ruptured ovarian cyst. The remaining five women presented with abdominal pain (n = 2), thyrotoxicosis (n = 1), menstrual irregularities/galactorrhoea (n = 1) and visual disturbances (n = 1). All women experienced intermittent pelvic pain during medical attendance. Pelvic ultrasound demonstrated enlarged multiseptated ovaries (volume ranging 27-442cm3). Ascites was noted in one woman. Six women had visual field defects due to optic chiasm compression on formal assessment. Median FSH was 26.10 u/L (8.3-33), but LH was &amp;lt;2.5 u/L in all cases. Estradiol (E2) far exceeded the reference range in 5/7 women (2990 to &amp;gt; 18000pmol/L);E2 was at the upper limit of normal in the remaining 2/7 women (960-1450pmol/L). Hyperprolactinaemia, hyperthyroidism and other pituitary hormones deficiency were noted in 6/7, 1/7 and 4/7 women respectively. All FGAs were macroadenomas with diameters ranging 16-48mm. Two patients were administered a somatostatin analogue prior to surgery, but FSH, E2 and tumour size did not change. Transsphenoidal surgery was performed in 6/7 women, and always improved symptomatic and biochemical features of OHSS; however, residual FGA tumour was present post-operatively in all cases studied. Limitations, reasons for caution It is possible that some ‘non-functioning’ gonadotroph adenomas cause subclinical problems including menstrual irregularity and mild OHSS which were never diagnosed. We have insufficient data to determine the prognosis for future pregnancy after FGA-induced OHSS. This study utilised historical case-notes, so some data is missing. Wider implications of the findings The ‘spontaneous’ presentation of OHSS may be confusing for clinicians. We report that FGA is an important cause of spontaneous OHSS which has well-defined biochemical and radiological characteristics, which may be treated effectively in the short-to-medium with pituitary surgery. Results of this study may provide greater awareness of FGA-induced OHSS. Trial registration number N/A

Recurrence and biochemical remission rates vary widely among different histological subtypes of pituitary adenoma. In this prospective study, we evaluated 107 consecutive primary pituitary adenomas operated on by a single neurosurgeon including 28 corticotroph, 27 gonadotroph, 24 somatotroph, 17 lactotroph, 5 null-cell and 6 plurihormonal. In each case, we performed direct endoscopic intraoperative inspection of the medial wall of the cavernous sinus, which was surgically removed when invasion was visualized. This was performed irrespective of tumor functional status. Medial wall resection was performed in 47% of pituitary adenomas, and 39/50 walls confirmed pathologic evidence of invasion, rendering a positive predictive value of intraoperative evaluation of medial wall invasion of 78%. We show for the first-time dramatic disparities in the frequency of medial wall invasion among pathological subtypes. Somatotroph tumors invaded the medial wall much more often than other adenoma subtypes, 81% intraoperatively and 69% histologically, followed by plurihormonal tumors (40%) and gonadotroph cell tumors (33%), both with intraoperative positive predictive value of 100%. The least likely to invade were corticotroph adenomas, at a rate of 32% intraoperatively and 21% histologically, and null-cell adenomas at 0%. Removal of the cavernous sinus medial wall was not associated with permanent cranial nerve morbidity nor carotid artery injury, although 4 patients (all Knosp 3-4) experienced transient diplopia. Medial wall resection in acromegaly resulted in the highest potential for biochemical remission ever reported, with an average postoperative day 1 GH levels of 0.96 ug/L and surgical remission rates of 92% based on normalization of IGF-1 levels after surgery (mean = 15.56 months; range 3–30 months). Our findings suggest that tumor invasion of the medial wall of the cavernous sinus may explain the relatively low biochemical remission rates currently seen for acromegaly and illustrate the relevance of advanced intradural surgical approaches for successful and durable outcomes in endonasal pituitary surgery for functional adenomas.

BackgroundFew studies to date have attempted to measure serum anti-Müllerian hormone (AMH) levels in adult men, and solid references ranges have not yet been defined in a large cohort.ObjectiveIn this study, we aimed, first, to establish the reference ranges for serum AMH and AMH-to-total testosterone ratio (AMH/tT) in adult males. Second, we investigated the relationship between serum AMH and both reproductive hormones and semen parameters.MethodsThis single-center retrospective study included 578 normozoospermic adult men. Serum AMH concentrations were determined with an automated sandwich chemiluminescent immunoassay.ResultsThe median serum AMH was 43.5 pmol/L. The 2.5th and 97.5th percentile values for serum AMH and AMH/tT were 16.4 and 90.3 pmol/L and 0.45 and 3.43, respectively. AMH was positively correlated with inhibin B and sperm concentration and negatively correlated with age, follicle-stimulating hormone (FSH), and progressive sperm motility. Interestingly, using immunofluorescence, we documented for the first time that AMH type II receptor (AMH-R2) is expressed in ejaculated human spermatozoa and gonadotrophic cells in the postmortem pituitary gland.ConclusionsWe establish a new age-specific reference range for serum AMH and AMH/tT. Moreover, AMH-R2 expression in human spermatozoa and gonadotrophic cells, together with the relationship between serum AMH levels and sperm motility or mean FSH levels, highlight new potential functions of AMH in regulating sperm motility or FSH secretion in adult men.

P62 is a protein adaptor for various metabolic processes. Mice that lack p62 develop adult-onset obesity. However, investigations on p62 in reproductive dysfunction are rare. In the present study, we explored the effect of p62 on the reproductive system. P62 deficiency-induced reproductive dysfunction occurred at a young age (8 week old). Young systemic p62 knockout (p62-/-) and pituitary-specific p62 knockout (p62flox/flox αGSUcre) mice both presented a normal metabolic state, whereas they displayed infertility phenotypes (attenuated breeding success rates, impaired folliculogenesis and ovulation, etc.) with decreased luteinizing hormone (LH) expression and production. Consistently, in an infertility model of polycystic ovary syndrome (PCOS), pituitary p62 mRNA was positively correlated with LH levels. Mechanistically, p62-/- pituitary RNA sequencing showed a significant downregulation of the mitochondrial oxidative phosphorylation (OXPHOS) pathway. In vitro experiments using the pituitary gonadotroph cell line LβT2 and siRNA/shRNA/plasmid confirmed that p62 modulated LH synthesis and secretion via mitochondrial OXPHOS function, especially Ndufa2, a component molecule of mitochondrial complex I, as verified by Seahorse and rescue tests. After screening OXPHOS markers, Ndufa2 was found to positively regulate LH production in LβT2 cells. Furthermore, the gonadotropin-releasing hormone (GnRH)-stimulating test in p62flox/flox αGSUcre mice and LβT2 cells illustrated that p62 is a modulator of the GnRH-LH axis, which is dependent on intracellular calcium and ATP. These findings demonstrated that p62 deficiency in the pituitary impaired LH production via mitochondrial OXPHOS signaling and led to female infertility, thus providing the GnRH-p62-OXPHOS(Ndufa2)-Ca2+/ATP-LH pathway in gonadotropic cells as a new theoretical basis for investigating female reproductive dysfunction.

The pituitary gland is the site of numerous neoplastic and inflammatory processes. The overwhelmingly most frequent tumors arise from cells of the anterior lobe, the pituitary neuroendocrine tumors (PitNETs). Immunohistochemistry assay staining for pituitary hormones is the core tool for classifying PitNETs, resulting in the diagnosis of somatotroph PitNETs, lactotroph PitNETs, and so on. For cases showing no hormonal expression, the updated WHO classification system now considers the assessment of several transcription factors: PIT-1 (pituitary-specific POU-class homeodomain transcription factor); T-PIT (T-box family member TBX19); and SF-1 (steroidogenic factor regulating gonadotroph cell differentiation) before rendering a diagnosis of null cell adenoma. Other tumors and disease processes of this site often mimic PitNETs radiographically and sometimes even clinically (ie, compression of the optic chiasm). These potpourri of processes include germ cell neoplasms (especially germinomas), tumors that originate from Rathke's pouch (craniopharyngiomas, Rathke's cleft cyst), tumors that originate from the posterior lobe of the pituitary (pituicytoma, spindle cell oncocytoma, granular cell tumor), and tumors that originate from the meninges (especially meningiomas). In addition to neoplasms, several described inflammatory and related conditions exist that need to be distinguished from PitNETs. These include lymphocytic hypophysitis and Langerhans cell histiocytosis, a neoplastic disorder of histiocytes. In this review, we aim to briefly describe the main pituitary and sellar lesions, with emphasis on the most common tumors, the PitNETs.

According to the WHO classification 2017 of Pituitary Tumors adenomas are classified not only by structure and immunostaining for pituitary hormones but also by expression of the pituitary transcription factors Pit-1, T-pit and SF-1. By these factors, three cell lineages can be identified: Pit-1 for the GH-, Prolactin- and TSH-cell lineage, T-pit for the ACTH-cell lineage, and SF-1 for the gonadotrophic cell lineage. By this principle, all GH and/or Prolactin producing and all TSH producing adenomas must be positive for Pit-1, all corticotrophic adenomas for T-pit, and all gonadotrophic for SF-1. In adenomas without expression of pituitary hormones immunostainings for the transcription factors have to be examined. If these are also negative the criteria for an endocrine inactive null cell adenoma are fulfilled. If one transcription factor is positive the corresponding cell lineage indicates a potential hormonal activity of the adenoma. So Pit-1 expressing hormone-negative adenomas can account for acromegaly, hyperprolactinemia, or TSH hyperfunction. T-pit positive hormone negative adenomas can induce Cushing's disease, and SF-1 positive hormone negative tumors indicate gonadotrophic adenomas. Instead of the deleted atypical adenoma of the WHO classification of 2004 now (WHO classification 2017) criteria exist for the identification of aggressive adenomas with a conceivably worse prognosis. Some adenoma subtypes are described as aggressive "per se" without necessity of increased morphological signs of proliferation. All other adenoma subtypes must also be designated as aggressive if they show signs of increased proliferation (mitoses, Ki-67 index>3-5%, clinically rapid tumor growth) and invasion. By these criteria about one third of pituitary adenoma belong to the group of aggressive adenomas with potentially worse prognosis. The very rare pituitary carcinoma (0.1 % of pituitary tumors) is defined only by metastases. Many of them develop after several recurrences of Prolactin or ACTH secreting adenomas. The correlation of clinical findings and histological classification of pituitary adenomas is very important since every discrepancy has to be discussed between clinicians and pathologists. Based on data of the German Registry of Pituitary Tumors a table for examinations of correlations is shown in this review.

The hypothalamus–pituitary–gonadal (HPG) axis is the endocrine regulation system that controls the woman’s cycle. The gonadotropin-releasing hormone (GnRH) plays the central role. In addition to the gonadotrophic cells of the pituitary, GnRH receptors are expressed in other reproductive organs, such as the ovary and in tumors originating from the ovary. In ovarian cancer, GnRH is involved in the regulation of proliferation and metastasis. The effects on ovarian tumors can be indirect or direct. GnRH acts indirectly via the HPG axis and directly via GnRH receptors on the surface of ovarian cancer cells. In this systematic review, we will give an overview of the role of GnRH in ovarian cancer development, progression and therapy.

Purpose: Epigenetic dysregulation plays a role in pituitary tumor pathogenesis. Some differences in DNA methylation were observed between invasive and noninvasive nonfunctioning gonadotroph tumors. This study sought to determine the role of DNA methylation changes in repetitive LINE-1 elements in nonfunctioning gonadotroph pituitary tumors. Methods: We investigated LINE-1 methylation levels in 80 tumors and normal pituitary glands with bisulfite-pyrosequencing. Expression of two LINE-1 open reading frames (L1-ORF1 and L1-ORF2) was analyzed with qRT-PCR in tumor samples and mouse gonadotroph pituitary cells treated with DNA methyltransferase inhibitor. Immunohistochemical staining against L1-ORF1p was also performed in normal pituitary glands and tumors. Results: Hypomethylation of LINE-1 was observed in pituitary tumors. Tumors characterized by invasive growth revealed lower LINE-1 methylation level than noninvasive ones. LINE-1 methylation correlated with overall DNA methylation assessed with HM450K arrays and negatively correlated with L1-ORF1 and L1-ORF2 expression. Treatment of αT3-1 gonadotroph cells with 5-Azacytidine clearly increased the level of L1-ORF1 and L1-ORF2 mRNA; however, its effect on LβT2 cells was less pronounced. Immunoreactivity against L1-ORF1p was higher in tumors than normal tissue. No difference in L1-ORF1p expression was observed in invasive and noninvasive tumors. Conclusion: Hypomethylation of LINE-1 is related to invasive growth and influences transcriptional activity of transposable elements.