bluuebunny/crossref_metadata_2025_split huggingface dataset:
New Transcriptional Insights into Silent and Active Corticotroph Pituitary Tumors at Single Cell Resolution
Authors: Zhang Dongyun , Hugo Willy , Bergsneider Marvin , Wang Marilene B , Kim Won , Vinters Harry , Heaney Anthony P 
Published: May 2021

Abstract
Silent pituitary corticotroph tumors derive from the Tpit (aka TBX19) pituitary lineage. Accounting for ~ 30% of corticotroph tumors, they are not infrequently clinically aggressive and invade locally into adjacent sellar structures, making complete surgical resection challenging and contributing to their higher recurrence rates. How silent and active corticotroph tumor subtypes differ is not clear although some studies reported that silent corticotroph tumors exhibit reduced PC1 expression causing impaired POMC processing. We used single cell RNAseq to compare the transcriptome between silent (n = 2) and active (n = 4) corticotroph tumors at the single cell level. We obtained an average of 265 million reads, and 24,682 genes per patient with an average of 1,240 genes expressed and 3,5442 unique molecular identifiers (UMIs) detected per cell. We further defined 5 distinct cell populations from a total of 23,269 cells, namely tumor cells (62%), stromal cells (25%), immune cells (7%), progenitor cells (5%), and a minor population of erythrocytes (1%). Tumor cells clustered in an origin-dependent manner and all expressed POMC and TBX19. However, the gene signatures of silent and active corticotroph tumors differed in 3 major aspects. Firstly, and supporting prior studies, a series of hormone processing peptidase genes, including PC1 (aka PCSK1), PDIA3, SEC11C, SPCS1 and CTSB, were reduced in silent corticotroph tumors (p=5.54e-5) compared to active corticotroph tumors. Secondly, genes involved in organization of secretory vesicles such as SCG5, TIMP1, VGF, SYT17, LGALS3 and CALY were also reduced in silent corticotroph tumors, which could further compound their inability to secrete ACTH. Thirdly, the silent corticotroph tumors exhibited several features of endothelial-to-mesenchymal transition (EMT), including increased expression of the mesenchymal genes CDH2 (aka NCAD), COL1A1, PCDH9, FGF5, ID2 (p=8.4e-3), and loss of EPCAM, which regulate cell migration and movement. Upstream analysis suggested that aberrant STAT3 activation may be related to these changes. Consequently, we noted that the stromal content was higher in silent corticotroph tumors (47.5% vs. 18.13%), concordant with the observed EMT de-differentiation of tumor cells. In summary, our scRNAseq analysis provides an unprecedented precise investigation of the transcriptomic features of thousands of heterogenous corticotroph tumor cells simultaneously. We demonstrate that although silent corticotroph tumor cells still express the pituitary lineage markers PITX1 and TBX19, they exhibit EMT, potentially affording increased migratory capacity at the cost of reduced neuroendocrine function with inability to produce and secrete mature ACTH. Our findings provide novel insights into the pathogenesis of silent versus active corticotroph tumor, but may reveal novel molecular targets for treatment of these challenging tumors.
The Secret Behind Anemia in Patients With Acromegaly
Authors: Firlatan Busra , Sendur Suleyman Nahit , Oguz Seda Hanife , Dagdelen Selcuk , Erbas Tomris A 
Published: May 2021

Abstract
Introduction: The GH/IGF-1 axis has regulatory effects on erythropoiesis. GH deficiency is associated with normocytic anemia in adults, and hemoglobin (Hb) concentrations are normalized after GH replacement. Little is known about the significance of anemia in acromegalic patients. The aim of this study is to evaluate the factors associated with anemia in patients with acromegaly.
Methods: A single-center retrospective cohort study was conducted. A total of 381 acromegalic patients who admitted to the University Hospital from 1980 to 2018 were included. Information regarding the demographic features, the pre-operative and follow-up hormone levels, radiological features of adenoma, treatment modalities, comorbidities, presence of cancer, colonoscopy results, medication history, presence and type of anemia together with the CBC, iron parameters, 25(OH)D, and vitamin B12 levels at the time of anemia detected were retrieved from the patient record system. Anemia was defined as Hb levels &lt;12.0 g/dL in women and &lt;13.0 g/dL in men.
Results: Of 381 acromegalic patients (120F/98M, age: 44±13yr/45±12 yr); 218 cases (57.2%) were diagnosed with anemia (67% normocytic, 33% microcytic) at a median duration of 6 months after the diagnosis of acromegaly. Hb values were 11.1±0.9 g/dL in women and 12.1±1.0 g/dL in men. Serum levels of ferritin and transferrin saturation were lower in anemic patients (p&lt;0.001). The levels of 25(OH)D and vitamin B12 were lower in the anemic group, but the differences were not statistically significant. Anemia was detected more likely in patients who had macroadenoma (p=0.001), suprasellar extension (p=0.001), and cavernous sinus invasion (p&lt;0.001) on pre-op MRI and residue tumor (p&lt;0.001) on post-op MRI. Patients with anemia had higher GH, IGF-1 and PRL levels (p&lt;0.01), and there was a significant difference related to the use of octreotide (p&lt;0.001). Anemia was observed more frequently in patients with a history of radiotherapy (29% vs. 10%, p&lt;0.001). Hypopituitarism was seen in 63 (29%) patients in the anemic group and 21 (13%) patients in the non-anemic group (p&lt;0.001). Cancer prevalence in the study population was 14.2%, and the most commonly diagnosed cancers were thyroid, bladder, prostate, and breast. Although not statistically significant, the prevalence of cancer was higher in anemic patients compared to non-anemic patients (17% vs. 10%). There was no significant difference regarding the presence of colorectal polyps between groups, and only two anemic patients were diagnosed with colon cancer.
Conclusion: This is the first study highlighting that anemia is a common comorbidity in patients with acromegaly, and that anemia can be related to characteristics of the tumor, hormonal status, type of treatment, and the presence of cancer in addition to nutritional factors. Therefore, a comprehensive evaluation is required for anemia in acromegalic patients.
Transcription Factor Immunohistochemistry in the Diagnosis of Pituitary Tumors
Authors: Lenders Nele F , Wilkinson Adam C , Wong Stephen J , Shein Tint T , Harvey Richard J , Inder Warrick J , Earls Peter E , McCormack Ann I 
Published: May 2021

Abstract
Objective: The clinical utility and prognostic value of WHO 2017 lineage-based classification of pituitary tumors have not been assessed. This study aimed to (1) To determine the clinical utility of transcription factor analysis for classification of pituitary tumors and (2) To determine the prognostic value of improved lineage-based classification of pituitary tumors. Methods: This was a retrospective evaluation of patients who underwent surgical resection of pituitary tumors at a tertiary referral centre between 1990 and 2016. Included patients were at least 18 years of age and had complete histopathological data, forming the “histological cohort”. Patients with at least 12 months of post-surgical follow up were included in the subgroup “clinical cohort”. The diagnostic efficacy of transcription factor immunohistochemistry in conjunction with hormone immunohistochemistry was compared with hormone immunohistochemistry alone. The prognostic value of identifying “higher risk” histological subtypes was assessed. Results: There were 172 patient tumor samples analyzed in the histological cohort. Of these, there were 96 patients forming the clinical cohort. Subtype diagnosis was changed in 24/172 (14%) of tumors. Within the clinical cohort, there were 21/96 (22%) patients identified with higher risk histological subtype tumors. These were associated with tumor invasiveness (p=0.032), early recurrence (12-24 months, p=0.016), shorter median time to recurrence (38 [IQR 20-68.5] v 15 [IQR 12-27.25] months, p=0.02) and reduced recurrence-free survival (p=0.023). Conclusions: Application of transcription factor analysis, in addition to hormone IHC, is associated with improved diagnostic information.
Single Nucleus Transcriptome and Chromatin Accessibility Landscapes of Human Pituitaries
Authors: Ruf-Zamojski Frederique Murielle , Zhang Zidong , Zamojski Michel , Smith Gregory R , Yianni Val , Willis Thea L , Mendelev Natalia , Pincas Hanna , Seenarine Nitish , Amper Mary Anne S , Vasoya Mital , Zhou Xiang , Gambino Luisina Ongaro , Schang Gauthier , Mofsowitz Sagie , Nair Venugopalan D , Welt Corrine Kolka , Troyanskaya Olga G , Turgeon Judith L , Bernard Daniel J , Andoniadou Cynthia Lilian , Sealfon Stuart C 
Published: May 2021

Abstract
The pituitary gland regulates key physiological functions, including growth, sexual maturation, reproduction, and lactation. Here, we present a paired single-nuclei (sn) transcriptome and chromatin accessibility characterization of six post-mortem human pituitaries. These samples were from juvenile, adult, and elderly male and female subjects. Well-correlated snRNAseq and snATACseq datasets facilitated robust identification of the major pituitary cell types in each sample. Using latent variable pathway analysis, we uncovered previously unreported coordinated gene expression modules and chromatin accessibility programs for each major cell type as well as an age-specific program across all the endocrine cell types. These largely appear to be congruent between human and mouse datasets. Given the importance of murine models in the study of human pituitary disorders and pituitary physiology, we next sought to compare expression profiles of pituitary cell types in mouse vs. human. Murine and human cell types were well correlated, exemplified by coordinated gene expression programs, especially for undifferentiated stem cells (SCs). In both species, we identified clusters corresponding to naive and committing SCs. All human SC clusters expressed the established SC markers SOX2 and SOX9, as well as genes involved in SC regulatory pathways (WWTR1, YAP1 andPITX2). Additional markers previously reported in murine pituitary SCs were also found in human SC, including WIF1, LGR5, FOS, CDH1, EGFR, LGR4, and WLS. Remarkably, in human, the main naive SC cluster was roughly divided into a high-JUN and a low-JUN expressing subgroup, whereas Jun expression was less pronounced in the murine SC cluster. In both species, committing SC clusters expressed the endocrine markers for POU1F1, TSHB, or POMC, while SCs committing to an intermediate lobe/melanotrope cell identity were distinguishable based on PAX7 expression. In addition, in the human datasets we identify a population of cells as originating from the pars tuberalis. We offer a range of markers that can be utilized for in vivo validation of these cells. Overall, the characterization of the murine and human pituitary SCs strongly suggests the co-existence of subpopulations with different lineage commitments in addition to a single uncommitted SC population. This sn atlas of the human pituitary is a valuable resource that will be made web-accessible.
Doppler Assessment of the Uterine Arteries for the Definition of Pubertal Onset in Girls
Authors: Cheuiche Amanda Veiga , da Silveira Letícia Guimarães , Lucena Iara Regina Siqueira , Puñales Marcia , Costenaro Fabiola , Kopacek Cristiane , Silveiro Sandra Pinho , de Paula Leila Pedroso 
Published: May 2021

Abstract
Pelvic ultrasonography (US) is a quick, non-invasive and low-cost method, and doppler analysis facilitates assessment of flow impedance measurement in the uterine vascular tree. The pulsatility index (PI) reflects blood flow impedance in the vessel distal to the sampling point and has been suggested as a parameter to define pubertal development. In order to evaluate the PI and US measurements of uterus and ovaries in girls with different pubertal stages, we performed a cross-sectional study in girls with normal pubertal development. US and Doppler assessed PI of the uterine arteries (defined as the difference between the peak systolic flow and end-diastolic flow divided by the mean maximum flow velocity), endometrial thickness, uterine and ovarian volumes were evaluated. All the US exams were performed with the same equipment by the same radiologist. Clinical data such as the age of menarche, pubarche and thelarche were recorded. Statistical analyses were performed in SPSS, with ANOVA test, Spearman correlation and ROC curve with Youden. One hundred and sixty-nine girls aged 5-16 years (mean 11.3 ± 1.8) who performed two hundred and two pelvic US were included (Tanner 1=20%, Tanner 2=22%, Tanner 3=23%, Tanner 4=17%, Tanner 5=17%). Mean age of thelarche, pubarche and menarche were 11.1 ± 1.8, 10.2 ± 1.2 and 12.2 ± 1.1 years respectively. Prepubertal girls (Tanner 1) had mean PI significantly higher than girls in initial puberty (Tanner 2 and 3 grouped) and in late puberty (Tanner 4 and 5 grouped), respectively 6.5 ± 2.27 vs. 4.15 ± 1.55 vs. 2.82 ± 1.06, p&lt;0.001 for all the comparisons. ROC curve analysis demonstrated that the PI is able to identify the onset of puberty with an area under the curve of 0.80 ± 0.04, P&lt;0.001, and a cutoff point of IP=5.05 presented a sensitivity of 0.77 and a specificity of 0.80 to identify the onset of puberty. When we combined the cutoffs of IP &lt;5.05 plus uterine volume &gt;3.75 cm³, we found a sensitivity of 0.72 and specificity of 0.90 to detect puberty. We identified a strong negative correlation between PI and uterine volume (rs=-0.72, p&lt;0.001) and a moderate negative correlation with endometrial thickness (rs=-0.68, p&lt;0.001) and right (rs=-0.60, p&lt;0.001) and left (rs=-0.59, p&lt;0.001) ovarian volumes. In conclusion, we found a significant reduction of the PI during pubertal development, reflecting a progressive increase in blood flow to the uterus, which can be a valuable non-invasive and highly specific tool to confirm the onset of puberty.
Effects of L-3,4-dihydroxyphenylalanine (L-Dopa) Treatment in the Neuroendocrine Response to Stress
Authors: Orrillo Santiago Jordi , Imsen Mercedes , Lizarraga Alfonsina , Romero Ana Clara , De Fino Fernanda , Zanoni Milagros Peña , Machado Alejandra Inés Abeledo , Zarate Sandra Cristina , Ferraris Jimena , Diaz Graciela Susana , Pisera Daniel 
Published: May 2021

Abstract
Stressful stimuli evoke a complex response mediated by two systems: the Sympathetic-Adreno-Medullar (SAM) axis and the Hypothalamus-Pituitary-Adrenal (HPA) axis. Among the factors involved in stress, glucocorticoids and catecholamines secreted from the adrenal glands and sympathetic nerves are the main effectors of the physiological adaptations to stressors. Besides these, prolactin (PRL) is another hormone secreted under stress conditions. Catecholamines are synthesized from the hydroxylated precursor L-Dopa. This agent is commonly used for the treatment of Parkinson’s disease and it would act as a neurotransmitter per se. On the other hand, it has been suggested that HPA axis dysregulation is a potential risk factor for the development of depression. In line with this, several studies reported that L-Dopa treatment may alter the serum levels of ACTH, PRL, and glucocorticoids in parkinsonian patients and Parkinson’s disease animal models. In the present study, we determined whether the chronic treatment with L-Dopa altered the stress response inducing depressive-like behaviours. Adult male Wistar rats were treated orally during 24 days with LEBOCAR® - commercial formulation of L-Dopa (75 mg/day) and Carbidopa (7.5 mg/day) - in drinking water. Animals were stressed by immobilization during the last 9 days of treatment and depressive-like behaviours were assessed by the sucrose intake and forced swimming tests. Behavioural tests showed no signs of depressive-like behaviours in the LEBOCAR®-treated and/or stressed rats. We next explored the SAM axis reactivity. Circulating noradrenaline and adrenaline increased in rats treated with LEBOCAR® (p&lt;0.05; HPLC). Also, the adrenals from stressed animals showed higher content of adrenaline (p&lt;0.05). Then, we studied the HPA axis activity. Chronically stressed rats displayed a lower ACTH secretion (ELISA) and a downregulation of POMC expression (qPCR) in the anterior pituitary (p&lt;0.05). In addition, LEBOCAR® treatment induced a reduction in serum ACTH and POMC levels (p&lt;0.05). As expected, serum corticosterone (ELISA) enhanced under chronic stress, an effect that was inhibited by treatment with LEBOCAR® (p&lt;0.05). Finally, pituitary PRL gene expression (qPCR) was downregulated by LEBOCAR® treatment with a more pronounced effect when rats were also stressed (p&lt;0.05). Our results suggest that L-Dopa alters the neuroendocrine stress response enhancing SAM axis reactivity and reducing HPA axis activity and PRL expression.
Infigratinib in Children With Achondroplasia (ACH): Design of PROPEL2 - A Phase 2, Open-Label, Dose-Escalation and Dose-Expansion Study
Authors: Savarirayan Ravi , Kannu Peter , Dambkowski Carl L , Rogoff Daniela , Irving Melita 
Published: May 2021

Abstract
Background: ACH, the most common non-lethal form of skeletal dysplasia, is characterized by defective endochondral ossification resulting from gain-of-function mutations in the fibroblast growth factor receptor 3 (FGFR3) gene, a negative regulator of endochondral bone formation. Current treatment options are non-targeted, ineffective, or painful interventions aimed at preventing or treating complications. Infigratinib is an orally bioavailable and selective FGFR1-3 tyrosine kinase inhibitor in development for FGFR-related conditions. In vitro data with infigratinib showed inhibition of FGFR1-3 activity with reversal of established growth arrest in chondrocytes. In vivo studies revealed dose-dependent improvements in foramen magnum and long bone length in Fgfr3Y367C/+ mice following treatment with infigratinib. Methods: PROPEL2 is a prospective, phase 2, open-label study of infigratinib in children with ACH. Children 3-11 years of age with ACH who have completed at least 6 months of observation in the observational PROPEL study are eligible to participate. PROPEL2 consists of dose escalation with an extended treatment phase, designed as dose finding, followed by a dose-expansion phase to confirm the selected dose and to provide evidence of efficacy. The primary endpoints of the dose-escalation/extended treatment phase are treatment-emergent adverse events and change from baseline in annualized growth velocity. Subjects (n=40) will be enrolled in ascending dose cohorts of approximately 10 subjects/cohort (4 cohorts planned) and treated for 6 months at their assigned dose, continuing for an additional 12 months with dose modifications as required. Up to 20 new subjects will be enrolled in the dose-expansion phase and receive infigratinib (mini-tablets, administered orally once daily) for 12 months. Secondary objectives include: safety/tolerability of infigratinib; changes from baseline in anthropometric parameters, including body proportions; and the pharmacokinetic/pharmacodynamic profile of infigratinib. An exploratory objective is evaluation of changes in ACH disease burden. Current Status: PROPEL2 is currently enrolling - the first subject was entered in July 2020. The planned total enrollment is 60 children with ACH (n=40 in the dose-escalation/extended treatment phase; n=20 in the dose-expansion phase). Following completion of PROPEL2, subjects have the opportunity to enroll in an open-label long-term extension study to assess the safety and efficacy of long-term administration of infigratinib in children with ACH.
Role of Genetic Analysis for the Differential Diagnosis of Delayed Puberty from the UK Puberty Cohort
Authors: Saengkaew Tansit , Dunkel Leo , Howard Sasha R 
Published: May 2021

Abstract
Distinction between self-limited delayed puberty (DP) and idiopathic hypogonadotropic hypogonadism (IHH) is challenging. Several biochemical tests have been applied to differentiate these conditions; however, all have limitations resulting in some patients undergoing inappropriate investigations and treatment. This study investigated whether the identification of a genetic defect in patients presenting with pubertal delay in adolescence through whole exome sequencing (WES) can help in distinguishing these two conditions. The WES data of 47 pubertal delay patients from Paediatric Endocrinology and Paediatric services around the UK were analysed. The variants were filtered for rare, predicted deleterious variants in genes from a virtual panel reported in IHH (n= 36), both IHH and self-limited (n=6), and self-limited DP (n= 4) that segregated with trait. Pathogenic variants were verified by Sanger sequencing, computational bioinformatics, and review of inheritance pattern as compared to previous reported mutations in this gene, and assigned a genotypic diagnosis as self-limited DP, IHH or overlapping. Full clinical history, physical examination and basic laboratory and hormonal investigations were used to determine the clinical diagnosis. Clinically, 38% of patients had self-limited DP and 26% had IHH and the rest of uncertain diagnosis as they were still undergoing a period of follow up under the age of 18 years. Twenty-four variants in 17 genes, accounting for 43% of the tested cohort, were identified to be potentially pathogenic variants, with most inherited in a heterozygous manner. We found a good correlation between genotypic diagnosis and phenotype diagnosis with Cohen’s kappa (k) = 0.235 (95% IC, 0.031 to 0.429). Thus, patients who carry variants reported only in self-limited DP with heterozygous inheritance were likely to have a clinical diagnosis of definite self-limited DP. On the other hand, patients carrying homozygous or loss of function variants in genes reported with heterozygous mutations in IHH will be very likely to have IHH. In addition, we identified variants in patients with clinical self-limited DP to have variants, which had been previously reported only in IHH, including DMXL2, CHD7, OTUD4, and SEMA3E. In summary, this study shows that there is a good genotype-phenotype correlation in patients presenting with pubertal delay. Therefore, genetic investigation may help to distinguish these two conditions, resulting in more accurate diagnosis and appropriate treatment modalities. In addition, new genes, previously implicated only in IHH, have been identified in self-limited DP, which indicate that these two conditions share similar disease mechanisms.