bluuebunny/crossref_metadata_2025 huggingface dataset:
Type 2 Diabetes Mellitus Remission in Obese Patients Under Bariatric Surgery: The Role of Preoperative Triglycerides Levels
Authors: Gonçalves Vanessa Guerreiro , Maia Isabel , Mendonça Fernando , Silva Maria Manuel , Neves João Sérgio , Pedro Jorge Pires , Salazar Cristina Daniela , Ferreira Maria João Silva , Costa Cláudia Fernandes , Canha Marta Borges , Lau Eva , Freitas Paula , Carvalho Davide M
Published: May 2021

Background: There is abundant evidence reporting the relationship between triglycerides levels and type 2 diabetes mellitus, however few studies confirmed the influence of triglycerides levels on the incidence of diabetes. Bariatric surgery may lead to the remission of type 2 diabetes mellitus, but the effect of basal serum triglycerides levels on this reversal is unknown. This study aimed to assess the association between preoperative triglycerides levels and pre- and type 2 diabetes mellitus remission in obese patients one year after bariatric surgery.

Methods: A retrospective study was conducted among 1959 obese patients who underwent bariatric surgery in our entre. Data on socio-demographic and clinical characteristics were used. Pre-diabetes and type 2 diabetes mellitus remission was defined as normal glycaemic measures at least one year’s duration without pharmacological therapy. Logistic regression models, crude and adjusted for sex, age, preoperative serum triglycerides levels and type of bariatric surgery, were used.

Results: The median of preoperative serum triglycerides level was 121.00 (SD=75.00) mg/dL, and 34.7% and 30.0% of patients presented pre- and type 2 diabetes mellitus, respectively, at baseline. Preoperative serum triglycerides levels were higher in type 2 diabetes mellitus patients (139.00 vs. 106.00 mg/dL, in normal glycaemic patients; p<0.001) and showed to be significantly correlated with fasting glycaemia, glycated

haemoglobin, homeostasis model assessment of insulin resistance and homeostasis model assessment of β-cell function. One year after bariatric surgery, 62.5% of patients with pre- or type 2 diabetes at baseline showed remission. Preoperative serum triglycerides levels were negatively associated with this remission (OR: 0.997; 95%CI=0.995–0.998), independently of, sex, age and type of bariatric surgery. Conclusions: One year after bariatric surgery, 62.5% of patients showed pre- or type 2 diabetes mellitus remission, being the preoperative serum triglycerides levels an important clinical parameter for remission.

Keywords: triglycerides, diabetes, remission, obesity, bariatric surgery
Activin A Plays a Critical Role in Adipose Tissue Wasting in the Progression of Cancer Cachexia
Authors: Luan Yi , You Mikyoung , Xu Pauline C , Thompson Tom , Kim So-Youn
Published: May 2021

Background: Nearly 50% of cancer patients suffer from cancer cachexia, a wasting syndrome with atrophy of white adipose tissue (WAT) and skeletal muscle. Cachexia leads to negative energy balance, limits cancer therapies, and reduces survival rate. It is characterized by body weight loss due to negative nutrients and energy balance from involuntary reduced food intake and abnormal metabolic conditions such as insulin resistance and hypertriglyceridemia. Cancer-driven factors such as activin A and IL-6 (interlukein-6) contribute to the occurrence of cachexia symptoms during cancer progression. While the importance of muscle atrophy has been emphasized in cachexia research, the underlying mechanism of adipose tissue wasting remains unclear. One proposed theory is that WAT switches to brown adipose tissue (BAT), characterized by the high expression level of UCP1 (uncoupling protein 1). Hypothesis: We hypothesize that activin A plays a critical role in adipose tissue wasting during cancer cachexia progression. Experiment: GDF9-iCre+; PIK3CA* female mice which shows cachexia symptoms in cancer progression were sacrificed before and after cachexia development. In addition, we injected FST288, an antagonist to activin A, for two weeks during cancer cachexia development. We harvested and analyzed multi-sites adipose tissues (gonadal, subcutaneous, interscapular and perirenal), muscle and liver. Serum activin A and IL-6 were measured using ELISA kits. DEXA and calorimetry analyses were performed, as well as immunohistochemistry, qPCR and western blotting assay. Results:GDF9-iCre+; PIK3CA* female mice started to display bilateral ovarian tumors around postnatal day (PD) 60, lose body weight around PD70 and became cachexia condition around PD80 with an increased level of serum activin A. Along with that, other body organs including liver, pancreas, muscle, and adipose tissues became dramatically small in mass. Our data proved that cachexia progression is correlated with the level of activin A rather than IL-6 in serum of GDF9-iCre+; PIK3CA* female mice. As serum activin A increased, adipocytes lost lipids and had distinct browning phenotypes in some adipocytes within WAT. Interestingly, calorimetry analysis did not display an increase in energy expenditure in cachectic mice although browning was evident in WAT. However, treatment with FST288 during cancer progression kept body weight and WAT in GDF9-iCre+; PIK3CA* female mice. Most of all, FST288 protected the size and lipid droplets of adipose tissues against WAT wasting during cachexia development. Conclusion: The progression of cancer cachexia impacts adipose tissues. Injection of FST288 supports the key role of activin A in the progress of cachexia. FST288 prevented adipose tissue wasting and cachexia development, revealing another evidence of the efficacy of activin A antagonist in preventing cancer cachexia development.
Exposure to the Widely Used Pyrethroid Pesticide Deltamethrin, Does Not Exacerbate High Fat Diet Induced Obesity or Insulin Resistance in C57BL/6J Mice
Authors: Tsakiridis Evangelia Evelyn , Morrow Marisa , Llanos Andrea , Wang Bo , Holloway Alison , Morrison Katherine Mary , Steinberg Gregory R
Published: May 2021

Deltamethrin is a commonly used pesticide for the control of mosquito populations. Despite widespread use, the effects of deltamethrin on adiposity and glucose homeostasis have been equivocal with some studies showing increased, decreased and no effect on adiposity and glycemic control. However, no study to date has investigated the effect of deltamethrin in mice housed at thermoneutral temperatures, which is important for modelling metabolic diseases in rodents due to reduced thermal stress and constitutive activation of brown adipose tissue. In the current study we demonstrate for the first time that deltamethrin reduces uncoupling protein-1 expression in brown adipocytes cultured in vitro at concentrations as low as 1pm. Meanwhile, in-vivo deltamethrin does not appear to alter glycemic control or promote adiposity at exposures equivalent to 0.01, 0.1 or 1.0 mg/kg/day. Together, our study demonstrates environmentally relevant exposure to deltamethrin does not exacerbate diet induced obesity or insulin resistance.
Inhibition of Angiopoietin-Like 3 (ANGPTL3) Reduces Adipose Tissue Insulin Resistance in Patients With Familial Partial Lipodystrophy
Authors: de Freitas Maria Cristina Foss , Akinci Baris , Neidert Adam , Hench Rita , Oral Elif A
Published: May 2021

Familial partial lipodystrophy (FPLD) is a rare disease characterized by selective loss of peripheral subcutaneous fat, usually affecting the trunk and limbs, but preservation in other areas, such as the face and neck. It is usually associated with dyslipidemia and diabetes mellitus, and currently, there are no approved specific therapies for this disease in the US. Reductions in circulating levels of ANGPTL3 either by homologous loss-of-function mutations in humans or by pharmacological inhibition in rodents are associated with reductions in triglyceride (and other atherogenic lipid) levels and protect from atherosclerosis, making it an attractive target for patients with FPLD and metabolic dyslipidemia. We performed a proof-of-concept study to assess the early efficacy and safety of targeting ANGPTL3 via antisense oligonucleotide ISIS-703802 (vupanorsen) in a small number of patients with FPLD. Four patients with FPLD (3F/1M; age range: 39–48; 1 with LMNA R482Q, 1 with LMNA R584H, and 2 with no causative genetic variant), diabetes (HbA1c>6.5%) and hypertriglyceridemia (>250 mg/dL at screening) were included. Patients received the study drug at a subcutaneous dose of 20 mg weekly for 26 weeks. The primary endpoint was the change in triglycerides at week 27. Other end-points of interest measured at the same time points included insulin secretion, sensitivity, lipid and hormonal changes in response to a 5 hour long mixed meal test and body composition measured by dual energy absorptiometry (DEXA). Treatment resulted in a 59.9±26.3 (mean±SD) % of reduction in triglycerides, 54.7±9.8% of reduction in serum ANGPTL3 levels and 50.8±27.4% of reduction in ApoCIII. Treatment with vupanorsen led to a reduction of 209.3±120.4 in adipose tissue insulin resistance (ADIPO-IR) from a baseline of 470.3±114.3 and the area under the curve (AUC) for circulating free fatty acid levels were decreased by 32.1±21.4 mmol/L/min from a baseline of 215.8±55.2 mmol/L/min. Glucose AUC and triglyceride AUC also decreased after treatment (-14.0±5.2 and -60.1±26.5 mg/dL/min, respectively). Analyzing body fat distribution using DEXA, we observed that the fat mass index (FMI) and trunk mass index (TMI) did not change from baseline, but the ratio of total fat mass/ fat mass from limbs decreased by 10.7±12.2. These data show a tendency for redistribution of central body fat to limbs. There were numerous adverse events observed that were related to common serious complications associated with diabetes and FPLD. Although limited, these results suggest that targeting ANGPTL3 with vupanorsen in patients with FPLD may have a therapeutic role by addressing multiple problems.
Protocol Based Standardized Endocrinological Evaluation of Children With Traumatic Brain Injury: A QI Initiative
Authors: Bhat Jayalakshmi Narayan , Gomez Ricardo , Schultz Scott
Published: May 2021

Introduction: Traumatic brain injury (TBI) can disrupt the hypothalamo-pituitary axis, causing central neuroendocrine dysfunction and hormone abnormalities (HPAD). As one-third of children can develop post-traumatic HPAD, a longitudinal and thorough follow up may be required to exclude the transient or late-onset HPAD.

Objective: To implement a protocol for standardized evaluation of children with TBI for the diagnosis of HPAD and to effectively establish a regular inpatient endocrine consultation and outpatient longitudinal follow up. Method: The study was divided into pre-QI (baseline phase) and QI phase (post-intervention phase). During the pre-QI phase, retrospective data were collected on children admitted with TBI at our institution for 1 year. The prevalence of HPAD and the percentage of children longitudinally followed in endocrine clinic were estimated. A consensus-based guidance protocol, detailing clinical and hormonal assay-based evaluation at presentation and during the follow up were formulated and implemented. Prospective data collection will be performed to estimate outcome measures (prevalence of HPAD, rate of initial endocrine consultation and endocrine outpatient follow up) and process measure (protocol adherence rate). Result: During the baseline phase (pre-QI), a total of 27 children, aged ≤19 years were admitted in the year for TBI management. The median (IQR) age at TBI diagnosis was 9 (3, 15) years. Motor vehicle accident was the predominant cause, accounting for 60%. In 85% of patients, the TBI was classified as severe based on GCS. Overall, only 8 children (30%) underwent limited (non-consultation based) endocrine evaluation (7 for central DI and 1 for central hypothyroidism) and 1 patient had complete evaluation (endocrinologist consulted). During the baseline period, the prevalence rate of transient central DI was diagnosed in 1 patient (4%). Implementation of protocol and post-intervention data collection are pending. Conclusion: The lower prevalence rate of HPAD in the current cohort of TBI patients may be due to under evaluation for endocrine dysfunction. QI initiative incorporating standardized evaluation using guiadance protocol will improve identification follow up rates of patients with endocrine dysfunction following TBI.
The Effects of Naphthenic Acids on Tryptophan Metabolism and Peripheral Serotonin Signalling
Authors: Jamshed Laiba , Perono Genevieve A , Jamshed Shanza , Cheung Kim Ann , Thomas Philippe J , Holloway Alison
Published: May 2021

Introduction: Serotonin produced in the periphery has been shown to affect glucose and lipid homeostasis. The availability of the amino acid tryptophan, the precursor of serotonin, affects serotonin availability. In addition, the metabolism of tryptophan via the kynurenine pathway produces physiologically active metabolites which have been shown to be altered under conditions of increased adiposity and dysglycemia. There is now evidence demonstrating some environmental xenobiotics, known to affect glucose and lipid homeostasis, can also alter serotonin production and key components of the kynurenine pathway. Recent evidence suggests that exposure to compounds present in petroleum and wastewaters from oil and gas extraction sites can impact endocrine signaling and result in aberrant lipid accumulation and altered glycemic control. However, whether any of these changes can be causally ascribed to altered serotonin synthesis/signaling or tryptophan metabolism remains unknown. The goal of this study was to determine the effects of exposure to naphthenic acid (NA), a key toxicant found in wastewater from bitumen (thick crude oil present in oil sands deposits) extraction on the enzymes involved in tryptophan metabolism and serotonin production.

Methods: McA-RH7777 rat hepatoma cells, were exposed to a technical NA mixture for 48 hours at concentrations within the reported range of NA found in wastewaters from oil extraction. We assessed mRNA expression for key rate-limiting enzymes involved in tryptophan metabolism that lead to either serotonin [Tph1] and/or kynurenine [Ido2 and Tdo2] production, as well as downstream enzymes in the kynurenine pathway [Afmid, Kyat1, Aadat, Kyat3, Kmo, Haao, Acmsd, Qprt]. We also examined the effects of NA on prostaglandin synthesis [Ptgs1, Ptgs2, Ptges] and signalling [Ptger2, Ptger4] as prostaglandins have been shown to be induced by serotonin and are linked to hepatic fat accumulation.

Results: NA treatment significantly increased Tph1 and Ido2 expression; this occurred in association with a significant increase in the expression of the inducible prostaglandin synthase Ptgs2 (COX-2), prostaglandin E synthase Ptges, and prostaglandin receptors Ptger2 and Ptger4. Acmsd was the only downstream enzyme in the kynurenine pathway that was significantly altered by NA treatment.

Conclusion: These results provide proof-of-concept that compounds associated with oil sands extraction have the potential to perturb key components of serotonin synthesis (Tph1) and tryptophan metabolism (Ido2, Acmsd). Furthermore, we found that the increase in Tph1 expression paralleled expression of Ptgs2. As increased prostaglandin production has been reported in association with nonalcoholic steatohepatitis, these data provide a potential mechanism by which exposure to NA and other petroleum-based compounds may increase the risk of metabolic disease.
Inhibition of CXCR2 by Glucocorticoids in Adipose Tissue
Authors: Amatya Shripa , Dhaibar Hemangini , Petrillo Maria Grazia , Cidlowski John A , Topete Diana Cruz
Published: May 2021

Obesity-induced chronic adipose tissue inflammation is a significant risk factor for metabolic and cardiovascular disease (CVD), which affects 30.3 million adults in the United States. Interaction of adipocytes with hormonal, metabolic and immune systems play an integral role in the underlying pathophysiological mechanisms that leads to development of obesity-related complications. Despite this association, the mechanisms that coordinate the inflammatory mediators in causing adipose tissue inflammation are not well understood. Glucocorticoids (GC) are well known for their potent anti-inflammatory actions; however, the mechanism by which GC coordinate the inflammatory response of adipocytes are unknown. From our genome-wide microarray data derived from adipocyte-specific glucocorticoid receptor (GR) knockout (AdipoGRKO) mice, we found that GR inactivation leads to a significant increase in pro-inflammatory gene in white adipose tissue (WAT). Additionally, WAT isolated from AdipoGRKO mice showed significant increase in immune cell infiltration, which correlates with our gene expression data. Among the top up-regulated genes, we found the C-X-C Motif Chemokine Receptor 2 (Cxcr2), which is a powerful mediator of chemotaxis to the sites of inflammation. Although studies have shown the presence of Cxcr2 in adipocytes and suggested the contribution of Cxcr2 signaling in adipocyte development, its role in integrating adipose tissue inflammatory response is unknown. This led us to hypothesize that GR is critical to repress Cxcr2 gene expression and its pro inflammatory effects in adipocytes. Our in vitro studies using 3T3-L1 cells derived adipocytes showed that treatment with the synthetic glucocorticoid, Dexamethasone (Dex) led to a significant repression of Cxcr2 mRNA and protein levels. Furthermore, these effects are mediated by GR acting directly to repress Cxcr2 gene expression. Systemic administration of corticosterone significantly altered Cxcr2 expression in adipose tissue compared to untreated mice further supporting our results. Together our findings suggest that administration of glucocorticoids could inhibit adipose tissue inflammation and alleviate the comorbidities that arise from inflamed adipose tissue.
A Novel Algorithm for Rare Disease Gene Prediction Based on Phenotypic Similarity
Authors: Fan Yibo , Flannick Jason
Published: May 2021

Genetic studies have yielded only a limited number of genes clearly implicated in endocrine disorders, in large part due to two current knowledge gaps. First, genome wide association studies (GWAS) of common diseases have yielded many associations that are hard to translate to causal genes and pathways. Second, whole exome sequencing (WES) studies have transformed diagnosis of rare diseases but often yield many variants of unknown significance that cannot yet be reliably prioritized for disease causality. We hypothesized that phenotypically similar diseases are more likely to share causal genes and pathways. Thus, genes implicated in a (rare or common) disease should be strong candidates to also contribute to a phenotypically similar disease. To test this hypothesis, we aggregated genes (a) for 3,209 rare diseases from OMIM and (b) nearby GWAS signals for 2,316 common diseases from the NHGRI/EBI GWAS catalog. We measured phenotypic similarity based on proximity in the Experimental Factor Ontology (EFO). Across ~2.7M common disease pairs, the number of genes shared increased with phenotypic similarity (Spearman p < 0.1). Similarly, across ~7.4M common and rare disease pairs and ~5.1M rare disease pairs, phenotypic similarity was significantly higher for disease pairs with at least one shared gene compared to those with no shared genes (T-test p < 0.05). We next developed an algorithm to predict genes for a rare disease based on its phenotypic similarity to other diseases and their known genes. Given a rare disease, the algorithm (a) identifies nearby diseases in the EFO; (b) collates their known genes and groups them into gene ontology (GO) terms; and (c) predicts the genes that occur in the most frequently observed GO term as potentially novel disease genes. We evaluated algorithm performance via cross-validation on rare diseases in OMIM. Across 140 rare endocrine diseases, the algorithm predicted on average 4.84 candidate genes with the correct (known but hidden by cross-validation) disease gene within the candidates 23.6% of the time; performance (5.11 candidates, 13.1% success rate) was similar for the other 3,069 rare diseases in OMIM. Examples include Leprechaunism (known gene INSR), for which genes INSR and TWIST2 were predicted based on phenotypic similarity to diseases Barber-Say syndrome, Rabson-Mendenhall syndrome and Gingival fibromatosis-hypertrichosis syndrome. Lubinsky syndrome (no known genes), for which genes ABCD1, LMNA, CNBP were predicted based on phenotypic similarity to diseases Ricker syndrome, X-ALD, DM1, Malouf syndrome, and Noonan syndrome. These data suggest that known phenotypic relationships and disease-gene databases can increase our ability to predict novel genes for less well-studied diseases, potentially speeding the biological translation of GWAS associations for common diseases and increasing the diagnostic yield of WES for rare diseases.
CAPA Neuropeptides Inhibit VA Activity in Malpighian Tubules of the Mosquito, Aedes Aegypti
Authors: Sajadi Farwa , Paluzzi Jean-Paul
Published: May 2021

Haematophagus insects, such as the female Aedes aegypti mosquito, face the challenge of excess ion and water intake after engorgement on a blood meal. To cope with this, adult female A. aegypti have a specialized excretory system that includes the Malpighian tubules (MTs), which are under rigorous control by several neuroendocrine factors to regulate transepithelial movement of ions/water. Produced in the CNS, the mosquito anti-diuretic hormone is a member of the CAPA peptide family, which share homology to the vertebrate neuromedin U peptide. CAPA peptides inhibit fluid secretion of MTs stimulated by select diuretic factors, 5HT and DH31 through the NOS/cGMP/PKG pathway. However, the anti-diuretic signalling mechanism and downstream cellular targets, such as ion channels and transporters, remain unclear. To study whether the V-type H+-ATPase (VA) plays a role in CAPA inhibition, we performed fluid secretion assays in MTs treated with diuretics and bafilomycin, a known VA inhibitor. Bafilomycin significantly inhibited DH31-stimulated fluid secretion 30 min post treatment compared to diuretic controls (p<0.05, n=22). Similar, however, delayed responses were seen in 5HT-stimulated MTs, while no affect was observed in DH44-stimulated secretion. An indirect way to measure whether CAPA inhibits VA activity was to measure the pH of the secreted fluid from diuretic-stimulated MTs treated with CAPA. In DH31-stimulated MTs supplemented with CAPA, there was an immediate significantly higher pH at 40 min, increasing up to 7.73±0.038 compared to control, 7.56±0.038 (p=0.0007, n=20). The pH of 5HT-stimulated MTs treated with CAPA was seen to significantly increase up to 7.75±0.061 (p=0.03, n=10) at the 60 min mark, in agreement with the delayed response previously seen. Unlike the effects observed with DH31 and 5HT, CAPA did not alter the pH of the secreted fluid in DH44-stimulated MTs. Alkalization of the secreted fluid in response to CAPA suggests inhibition of the proton pump, which may lead to constrained cation entry across the apical membrane of the MTs. To understand Na+/K+-ATPase (NKA) and VA activity in response to CAPA, we performed activity assays in diuretic-stimulated MTs. Adult female MTs treated with DH31 resulted in an increase of both NKA and VA activity compared saline controls. As expected, MTs incubated with both DH31 and AedaeCAPA-1 had a lower NKA and VA activity (p<0.05) resulting in activity levels comparable to saline control levels. The results thus far could suggest a novel mechanism for CAPA inhibition, blocking the VA to hinder fluid secretion. Investigating the pathway of CAPA inhibition and its role in countering diuresis will help provide a deeper understanding of the critical process of diuresis and its signaling mechanism.
Effects of Maternal Sucrose Consumption on Inflammation and Steroids in the Placenta and Fetal Brain
Authors: Seib Desiree R , Kachkovski George V , Bodnar Tamara S , Collier Abby C , Soma Kiran K
Published: May 2021

Consumption of sucrose (table sugar) is high in much of the world. The effects of a maternal diet high in sucrose on the placenta and fetal brain remain unknown. In rats, maternal consumption of sucrose at a human-relevant level has effects on the mother’s physiology and steroids, as well as long-lasting and sex-specific effects on the adult offspring’s brain and behavior. In the mothers, there are metabolic effects of sucrose intake, such as impaired glucose tolerance, increased liver lipids, and increased adipose inflammation. In rat dams, sucrose intake also decreases corticosterone levels in the blood but not in the brain. In the adult male offspring, preference for a high-sucrose diet and a high-fat diet increases due to maternal sucrose intake. In addition, maternal sucrose intake increases motivation for sugar rewards in a progressive ratio schedule of reinforcement in adult male offspring. In adult female offspring, corticosterone levels increase in the blood and brain as a result of maternal sucrose intake. In this study, we investigated the underlying mechanisms of the observed behavioral and endocrine effects in the adult offspring. Here, we examined cytokines and anti-inflammatory steroids in the placenta, amniotic fluid, and fetal blood and brain. In our model, we feed rat dams either a high-sucrose diet (26% of kCal) or an isocaloric, matched, control diet (1% sucrose) 10 weeks prior to and during gestation. At embryonic day 19 (E19), we collected maternal blood, placenta, amniotic fluid, fetal blood, and fetal brain. We use Palkovits punch to microdissect the placenta and fetal brain. Next, we use a liquid chromatography-tandem mass spectrometry (LC-MS/MS) assay, which is highly precise and specific, to measure multiple steroids (e.g. corticosterone, progesterone, estradiol, allopregnanolone). The method is highly sensitive, and we can measure neurosteroids in multiple regions of the fetal brain (e.g. prefrontal cortex, nucleus accumbens, hypothalamus, hippocampus). Moreover, we will examine steroidogenic enzymes and cytokines in the fetal brain and placenta. Preliminary data show distinct steroid patterns in amniotic fluid and fetal blood, as well as in different parts of the placenta.
Effects of Pre and Post-Natal Androgen Exposure on Psychosexual Aspects and Gender Change in Congenital Adrenal Hyperplasia Due to 21-hydroxylase Deficiency
Authors: Batista Rafael Loch , Inacio Marlene , de Miranda Mirela Costa , Madureira Guiomar , Gomes Larissa Garcia , Rodrigues Andresa De Santi , Bachega Tania A , Mendonca Berenice Bilharinho
Published: May 2021

Introduction: Congenital Adrenal Hyperplasia (CAH) is defined as a group of autosomal recessive disorders characterized by a deficiency of the enzyme required to synthesize cortisol by the adrenal cortex. Defects in the 21-hydroxylase enzyme make up 90% of CAH. These defects impaired cortisol synthesis leading to an ACTH increase resulting in androgen excess in both salt-wasting (SW) or simple virilizing (SV) forms. As androgens play a role in the human psychosexual development favoring male psychosexuality, this study was designed to evaluate the impact of androgen exposure on the psychosexuality of individuals with CAH due 21-hydroxylase deficiency. Methods: This retrospective cohort includes 46,XX individuals (115 female-assigned; 8 male-assigned) with a molecular diagnosis of CAH due to CYP21A2 pathogenic variants in homozygous or compounds heterozygous state. External genitalia virilization was scored using Prader scale. Phenotype, time at diagnosis, sex assignment, and gender change were assessed. The gender role at childhood was assessed through the playmates and toys profile at childhood. Gender identity was assessed by a projective psychological test (HTP). Sexual orientation was assessed by self-report sexual identity. Compliance of glucocorticoid replacement was assessed by adequate testosterone and androstenedione serum levels for age. Results: CAH was diagnosed at the neonatal time in 73% (n=78). Fifth-nine (51%) had the SW form and 49% (n=56) had the SV form. While all cases of SW were diagnosed at the neonatal time (0.12 ± 0.14 months), the mean age at diagnosis among SV was 6.03 ± 8.45 years (p=<.001). The median of Prader score was 3 in both forms. Male sex assignment was associated with more virilized external genitalia (p=.002). Gender change occurred in 6 cases (female to male), all with SV form. The prader score was higher among those who changed gender (p=.01). All of those who changed their gender had poor treatment compliance. A total of 13% (n=15) of all groups defined themselves as homosexual. There was a strong association between male toys and preference for male playmates in childhood with homosexuality and male gender identity in adulthood with both gender change from female to male and homosexuality. Conclusion: Prenatal androgen exposure favors male psychosexuality in 46,XX CAH individuals as observed by the association between highest Prader scores and all assessed psychosexual outcomes. This influence is also substantiated by post-natal androgen exposure as observed by compliance issues and late diagnosis among those who changed from female to male gender.
Interrupted IGF-1 Feedback in GHRH Neurons and Somatotrophs Results in Impaired Weight Gain and Increased Energy Expenditure
Authors: Al-Samerria Sarmed , Aloqaily Bahaa , Negron Ariel , Wondisford Fredric Edward , Radovick Sally
Published: May 2021

Growth hormone (GH) expression and release are thought to be primarily regulated by the counter-regulatory effects of growth hormone-releasing hormone (GHRH) and somatostatin (SST). Several reports generated in our lab and others suggest that there are other factors that regulate GH production, such as insulin-like growth factor 1 (IGF-I). Using GH-Cre recombinase targeting of the somatotroph-specific IGF-1R knockout (SIGFRKO) mouse model, we have previously demonstrated the role of IGF-1 signaling in negative feedback regulation of GH production. This model, however, presented with an incomplete phenotype, suggesting additional regulatory pathways in the hypothalamus. To provide insight into this mechanism, we have developed new transgenic mouse models that maintain the integrity of the hypothalamic-pituitary GH axis, with the single exception of IGF-IR deficiency in both hypothalamic GHRH neurons and somatotroph cells, termed GHRH-somatotroph IGF-1R knockout (G-SIFGRKO). Axiological assessments showed normal linear growth until week 14 of age, both male and female G-SIFGRKO mice presented with a significant reduction in growth velocity compared to control animals. Indirect calorimetry assessment performed at 12–14 weeks of age demonstrated that G-SIFGRKO mice had higher volume O2 consumption and lower volume CO2 production associated with increased energy expenditure than controls. The calculated respiratory exchange ratio was significantly reduced in G-SIFGRKO mice with no changes observed in either ambulatory or total activity. Furthermore, glucose and insulin tolerance tests showed no differences in glucose metabolism between G-SIFGRKO and controls. Collectively, these data provide further confirmation of the combinatorial role of IGF-1 signaling in regulating GH production and, for the first time, highlight a new GHRH-IGF-1R mediated pathway to regulate body growth and energy balance. Targeting this pathway has the potential to lead to a better understanding of the intersection between growth and metabolism and therapeutic approaches for obesity.
Identification of a Long Non-Coding RNA Derived From the Antisense Strand of the Thyroid Hormone Receptor Beta Gene
Authors: Ishii Sumiyasu , Horiguchi Kazuhiko , Amano Izuki , Yamada Masanobu , Koibuchi Noriyuki
Published: May 2021

Background: Long non-coding RNAs (lncRNAs) have various functions. Here, we describe a novel lncRNA which is induced from the antisense strand of the thyroid hormone receptor beta (THRB) gene. Methods: We sequenced RNA from leukocytes and identified a novel lncRNA derived from the antisense strand of the THRB gene. The lncRNA was named THRB antisense RNA 2 (THRB-AS2). Transcriptional regulation by THRB-AS2 was assessed by reporter assays. The putative translation initiation codons of THRB-AS2 were mutated to assess potential THRB-AS2 protein function. Results: THRB-AS2 stimulated the activities of multiple promoters irrespective of thyroid hormone response elements. Thyroid hormone treatment did not affect the function of THRB-AS2. These data suggest that THRB-AS2 is a potential general transcriptional activator and has broad specificity on target genes. Mutations in putative translation initiation codons of THRB-AS2 did not affect function, suggesting that THRB-AS2 acts as an RNA molecule. Conclusions: Although a small number of lncRNAs derived from antisense strand of the THRB gene, such as THRB-AS1, has been already reported, their functions remain largely unknown. To our knowledge, this is the first report of the functions of a lncRNA derived from the antisense strand of the THRB gene.
Universal Multi Gene Panel Testing For Individuals With Pheochromocytomas And Paragangliomas
Authors: Horton Carolyn , Richardson Marcy , Durda Kate , Yussuf Amal , Jackson Michelle , Jasperson Kory , Tian Yuan , LaDuca Holly , Else Tobias
Published: May 2021

Background: Pheochromocytomas (PCCs) and paragangliomas (PGLs) (PPGLs) are a genetically heterogeneous entity, with roughly 25-40% of cases found to harbor a pathogenic or likely pathogenic germline alteration. Existing practice guidelines advocating for the use of a sequential gene testing strategy to identify individuals with hereditary PPGL are driven by the presence of specific clinical features and predate the routine use of multigene panel testing (MGPT). Here we describe results of MGPT for hereditary PPGL in a clinically and ancestrally diverse cohort from a diagnostic laboratory. Methods: Demographic and clinical information of individuals undergoing targeted MGPT for hereditary PPGL were collected from test requisition forms and supporting clinical documents provided by the ordering clinician and retrospectively reviewed. Individuals underwent MGPT of 10-12 genes depending on test order date. From August 2013 through May 2015, 560 individuals had targeted MGPT that included 10 genes (NF1, MAX, SDHA/B/C/D/AF2, RET, TMEM127, and VHL), and from May 2015 through December 2019, 1167 individuals had panel testing of 12 genes due to the addition of MEN1 and FH. Results: Overall, 27.5% of individuals had a pathogenic or likely pathogenic variant (PV), 9.0% had a variant of uncertain significance, and 63.1% had a negative result. Out of all PVs, most were identified in SDHB (40.4%), followed by SDHD (21.1%), SDHA (10.1%), VHL (7.8%), SDHC (6.7%), RET (3.8%), and MAX (3.6%). PVs in FH, MEN1, NF1, SDHAF2, and TMEM127 collectively accounted for 6.5% of PVs. Clinical predictors of a PV included extra-adrenal location, diagnosis before the age of 45 years, multiple tumors, and positive family history (fhx) of PPGL. Affected individuals with a fhx of PPGL were the most likely to have a PV (70.6% of individuals with PCC + fhx; 85.9% of individuals with PGL + fhx). The positive rate in nearly all clinical subgroups even without predictors of a PV remained over 10%, including individuals with a single tumor (PCC = 16.7%; PGL = 46.7%) and those without a fhx (PCC and negative fhx = 15.8%; PGL and negative fhx = 43.7%). Restricting genetic testing of hereditary PPGL to only SDHB/C/D genes misses a third (31.8%) of individuals with PVs. Among individuals with PVs in syndromic genes, over half (41.5%) did not have any additional syndromic features beyond PPGL reported by the ordering clinician. Conclusion: Our data demonstrate a high diagnostic yield in individuals with and without established risk factors, a low inconclusive result rate, numerous individuals with syndromic PVs presenting with isolated PPGL, and a substantial contribution to diagnostic yield from rare genes when included in testing. These findings support updating practice guidelines to incorporate universal testing of all individuals with PPGL and the use of concurrent MGPT as the ideal platform.
Young Adult LEW.1WR1 Rats, a Model of Liver FAT10 Overexpression, Develop Insulin Resistance and Fatty Liver With Age
Authors: Wimalarathne Madushika M , Mercado Luis D , Vidal Quiana C Wilkerson , Wolfsberger James , McConnell Victoria J , Vogler Bernhard , Love-Rutledge Sharifa Tahirah
Published: May 2021

As human lifespan increases, comorbid conditions that impact quality of life have become a serious problem. FAT10 has been identified as a gene that when knocked out, improves age associated metabolic dysfunctions and increased longevity in mice (1). There is increased Fat10 expression in the liver in obesity (2,5). Providing evidence that fat10 expression may be important for triggering the transition to metabolic dysfunction in aging. Adult LEW.1WR1(1WR1) rats have increased body mass without excess abdominal fat mass compared to control rats (3). Yet, it was unclear where the excess mass was stored. We hypothesized that the 1WR1 rat would develop increased liver fat mass as a product of increased insulin resistance in response to increased liver fat10 expression over time. To test this hypothesis, we did insulin tolerance tests(7 weeks & 15 weeks), triglyceride assays, and histological analysis of the liver(23 weeks), in 1WR1 rats(n=7) and Wistar Furth (WF) rats(n=7) on control diets. We analyzed images using histological scoring for nonalcoholic fatty liver disease from the literature (4). We also assessed the slides for Mallory Denk bodies (MBs). The body mass of 1WR1 rats were increased compared to WF rat groups starting from the age of 7 weeks (391.4∓8.572g vs. 271.8∓11.62g; p <0.0006). 1WR1 rats became more insulin resistant with age, the 1WR1 rat group has increased AUC of 7 and 15 week Insulin Tolerance Tests (401.5∓23.54 vs. 245.3∓10.20 7w ITT1; p= 0.0728, 15w ITT2 328.2∓14.86 vs. 217.8∓9.; p <0.0003) compared to WF rats. 1WR1 rats have increased liver mass (11.85g∓0.7699g vs. 7.235g∓0.3864g; p=0.0006) liver triglyceride levels compared to WF rats (192.8∓21.8 mg/mL vs. 130.1∓13.075 mg/mL; p=0.0728). 1WR1 rats have increased steatosis scores(1.857∓0.2608 vs. 1.143∓0.1429;p= 0.0862) yet significantly reduced inflammatory foci level (2∓0.8165 vs. 3∓0;p= 0.007), most 1WR1 hepatocytes are enlarged (ballooned) and contained MBs compared to WF rats suggesting 1WR1 rats have already passed the early inflammation stage. Adult 1WR1 rats developed reduced insulin sensitivity and lipid accumulation in the liver. These data support our hypothesis that 1WR1 rats would develop increased liver fat and impaired insulin resistance in response to aging and show that this process may be inflammation driven.

(1) Canaan et al.,PNAS. April 2014; 111 (14): 5313-5318.(2)Vidal et al., FASEB.2020, 34: 1-1,(3) Collins et al., J Endocr Soc. 2019;3(1),(4).Kleiner et al., Hepatology, 2005; 41 (6): 1313–1321,(5).Dali-Youcef et al., Hepatol Commun. 2019;3(9):1205-1220.
Growth Hormone Secretion Does Not Increase After Interleukin-2 Administration in Healthy Men
Authors: Roelfsema Ferdinand , Yang Rebecca , Veldhuis Johannes D
Published: May 2021

Context: Interleukin-2 (IL2), a proinflammatory cytokine, is used for treatment of malignancies. Increased cortisol and ACTH were noted, but GH secretion was not investigated in detail. This is the first study in healthy men which uses moderate high IL2 doses as used in cancer treatment. Objective: The goal of this study was to quantify GH secretion after a single sc injection of IL2 in young and older healthy men in relation to dose, age and body composition. Design: This was a placebo-controlled, blinded, prospectively randomized cross-over study in 17 young subjects (mean age 24.1 yr) and 18 older subjects (mean age 63.9 yr). The subjects underwent 24 h of blood sampling at 10-min intervals, starting at 1800 h. At 2000 h IL2 (3 or 6 million units per m2 body surface) or saline was injected sc. Lights were off between 2300 and 0700 h. Outcome Measures: Deconvolution analysis of GH. Abdominal visceral fat (AVF) was calculated from single slice CT. Results: GH secretion (pulsatile and total) was negatively related to AVF (R=0.67, P<0.0001) and to age (R=0.56, P=0.001). These relations were maintained after IL2 administration. GH profiles were pulsatile under both experimental conditions. Since the effect of IL2 might be limited in time, GH deconvolution analyses were performed on the complete 24 h data series, the period with lights off and 6 h after IL2 administration. Total 24 h GH secretion decreased non-significantly from 74.6 ±10.2 to 67.5±7.4 µg/L (P=0.25) and pulsatile secretion from 69.4±9.8 to 62.2±7.4 µg/L (P=0.23). In the GLM procedure the effect of IL2 treatment was borderline significant (P=0.08), no interaction with age (P=0.28), but borderline with IL2 dose (P=0.07), and caused by decreased GH secretion (88 to 67 µg/L) at a low dose IL2. In a sub-analysis of the two age groups no effect on GH secretion in the older group was noted, in contrast to young subjects, especially on low dose IL2 (P=0.07). The analysis was refined further by calculating cumulative pulse masses in 2-h bins. By this approach it could be demonstrated that the cumulative GH pulse mass in older subjects across the 24 h was unchanged, while that in young subjects was temporarily inhibited in the bins between midnight and 0400 h (P=0.019 and 0.038). Conclusion: This study demonstrated that IL2 temporarily diminished GH secretion in young but not elderly volunteers. Pituitary hormone secretion by IL2 has not been studied as extensively as other cytokines, including IL1, IL6 and TNF. IL2 stimulates ACTH and TSH, inhibits LH and has no effect on GH secretion by rat pituitary (PNAS 58:185,1993). In patients with renal cancer IL2 infusion has no effect on GH levels (Anti-Cancer Res 10:753,1990. The present findings extend these observations to a large group of healthy volunteers, in whom we demonstrated previously suppression of the gonadal axis and activation of the HPA-axis (JCEM 101:539,2016, Endocr Connect 9:637,2010).
Pancreatic Ductal Adenocarcinoma Highly Expresses Activin A: Implications in Adipose Tissue and Cancer Cachexia
Authors: Xu Pauline , You Mikyoung , Luan Yi , Eldani Maya , Chung Soonkyu , Kim So-Youn
Published: May 2021

Background: Pancreatic ductal adenocarcinoma (PDAC) is currently the third leading cause of cancer death in the United States and is projected to become the second leading cause by the year 2030. Prognosis for patients with metastatic disease remains dismal. Cancer cachexia is seen in over 85% of PDAC patients who often have the most severe degrees of cachexia and experience adipose tissue loss prior to skeletal muscle loss early in the disease process. Several factors have been proposed to induce cachectic symptoms in human patients, including inhibin subunit βA, or activin A. Hypothesis: While muscle wasting has been the most frequently studied mechanism in cachexia research, changes in adipose tissue are increasingly understood as important components of body weight loss in cachectic patients. We hypothesized that there is a correlation between the marked increase in activin A production in PDAC patients and the remodeling of adipose tissue and consequent cancer-associated cachectic state. Experimental Design: We measured serum activin A levels of a cohort of PDAC patients and analyzed the expression of activin A in tumor-derived cell lines and biopsies of both humans and mice. We further investigated the effect of activin A on remodeling of adipose tissue secondary to tumor progression in PDAC patients and an orthotopic murine model. Results: We observed that PDAC cell lines express and secrete activin A. We recognized a loss of adipose tissue mass and adipocyte diameter in PDAC patients and our orthotopic PDAC mouse model in relation to increased circulating activin A. We also noted that both exogenous activin A and conditioned medium from pancreatic tumor-derived cell lines dampened adipocyte differentiation and lipid droplet formation via reduction of PPARγ expression in mouse mesenchymal stem cells. These treatment conditions also reduced lipid droplet size without upregulating traditional markers of adipose tissue browning and lipolysis such as UCP-1 and ATGL in mature mouse adipocytes. PPARγ, UCP-1, and ATGL expression are also heavily downregulated in adipose tissue of PDAC patients. Furthermore, our studies revealed that the expression of extracellular matrix proteins such as collagen I and fibronectin is dramatically upregulated in adipose tissue of PDAC patients and our orthotopic PDAC mouse model. Thus, we found that there is a clear correlation between elevated levels of activin A and the progression of cancer-associated cachexia in PDAC. Discussion: Our results reveal an imperative role of activin A in relation to the loss and remodeling of adipose tissue in the progression of cachexia in PDAC patients.
Safety Results From MPOWERED, a Phase 3 Trial of Oral Octreotide Capsules in Adults With Acromegaly
Authors: Freda Pamela , Fleseriu Maria , Dreval Akexander V , Pokramovich Yulia , Bondar Irina , Isaeva Elena , Huang Wenyu , Molitch Mark E , Macut Djuro P , Leonova Nina , Raverot Gerald , Gilgun-Sherki Yossi , Ludlam William H , Patou Gary , Haviv Asi , Gordon Murray B , Biermasz Nienke , Melmed Shlomo , Strasburger Christian J
Published: May 2021

Background: Injectable somatostatin receptor ligands (iSRLs) have been a mainstay in acromegaly treatment. Oral octreotide capsules (OOC; MYCAPSSA®) were recently approved in the United States. Results from the placebo-controlled CHIASMA OPTIMAL and open-label CH-ACM-01 studies showed an OOC safety profile consistent with that of iSRLs with no new or unexpected safety signals. Results of the MPOWERED trial have enabled a comparison of OOC safety and efficacy with iSRLs.

Methods: To enter MPOWERED, patients must have the following: acromegaly diagnosis, biochemical control of acromegaly (insulin-like growth factor I <1.3 × upper limit of normal; mean integrated growth hormone <2.5 ng/mL), and ≥6 months’ iSRLs treatment (octreotide or lanreotide). Eligible patients entered a 26-week Run-in phase to determine the effective OOC dose; responders at week 24 then entered a 36-week randomized controlled treatment (RCT) phase receiving OOC or iSRLs. Safety was monitored as adverse events (AEs) in both arms throughout the trial, including the RCT.

Results: In the RCT, incidence of treatment-emergent adverse events (TEAEs) was similar between groups; 39 patients (70.9%) in the OOC group and 26 (70.3%) in the iSRL group had ≥1 TEAE. 19 patients (34.5%) in the OOC and 15 (40.5%) in the iSRL group had treatment-related TEAEs. Occurrence was similar for serious AEs (OOC, 5.5%; iSRL, 8.1%) as well as TEAEs classified as severe (OOC, 9.1%; iSRL, 10.8%). One patient in the OOC group discontinued due to a TEAE. The most common gastrointestinal TEAEs were flatulence (OOC, 25.5%; iSRL, 21.6%), nausea (OOC, 20.0%; iSRL, 8.1%), diarrhea (OOC, 10.9%; iSRL, 13.5%), abdominal pain (OOC, 9.1%; iSRL, 8.1%), and constipation (OOC, 5.5%; iSRL, 13.5%). AEs of interest were infrequent, including cholelithiasis (OOC, n=0; iSRL, n=1 [2.7%]) and secondary hypothyroidism (OOC, n=1 [1.8%]; iSRL, n=0). In the iSRL group, 32.4% of patients reported injection site reactions (ISRs) during the RCT, and 47% of patients reported ISRs as part of the Acromegaly Treatment Satisfaction Questionnaire, a newly validated patient-reported outcome tool.1

Conclusion: Safety results from MPOWERED align with prior trials, showing that the OOC safety profile is consistent with that of iSRLs as well as the acromegaly disease burden. No new or unexpected safety signals were identified during the trial. Safety results were mostly similar between OOC and iSRLs, although patients in the OOC group did not experience any ISRs.

1Fleseriu M, et al. Pituitary. 2020 Aug;23(4):347-358.