bluuebunny/crossref_metadata_2025 huggingface dataset:
SAT148 A Rare Case Of Proinsulinoma In A Patient With Type 2 Diabetes Mellitus
Authors: Tirthani Ekta , Erenler Feyza
Published: October 2023

Disclosure: E. Tirthani: None. F. Erenler: None.

Background: A proinsulinoma is a very rare neuroendocrine tumor of the pancreas, which predominantly secretes proinsulin, the precursor hormone of insulin. Even though not as potent as insulin, proinsulin in higher amounts can bind to insulin receptors and result in hypoglycemia. Most patients present with initial hypoglycemia symptoms at the time of diagnosis of the tumor. The diagnosis becomes challenging in patients with underlying type 2 diabetes mellitus (T2DM) who may initially have a spontaneous remission of diabetes, followed by hypoglycemia episodes later in the disease course. Clinical Case: A 66-year-old man presented to our clinic in October 2020 with frequent episodes of dizziness and diaphoresis, which were relieved by eating food. The patient's symptoms began in 2018, occurred during the daytime, and were exacerbated by exercise. He had a past medical history of T2DM diagnosed in 2009 (HbA1c=10%) and treated with glipizide and metformin; however, over the years, his HbA1c improved, and glipizide was discontinued. By 2018 his HbA1c was down to 5.9%, and metformin was also stopped. The patient denied any change in his weight, diet, or exercise frequency during this time course. A fingerstick blood glucose (BG) checked during one of these episodes was 51 mg/dL. An abdominal CT scan in August 2020 revealed a 12x13 mm enhancing mass within the neck of the pancreas concerning for a neuroendocrine tumor. The patient was further evaluated with a 14-day continuous glucose monitor (CGM), which showed that the patient's BG was <70 mg/dL 31% of the time and <55 mg/dL 6% of the time. Additional fasting blood work showed a proinsulin level of 191.3 pmol/L (normal ≤18 pmol/L), insulin 15.5 uIU/mL (2-25 uIU/mL), BG 103mg/dL, C-peptide 2.44 ng/mL (0.80-3.85 ng/mL) and chromogranin A 124 ng/mL (25-140 ng/mL) which suggested the diagnosis of proinsulinoma. A Gallium-DOTATATE scan showed a localized tumor within the pancreas with no metastasis. The patient underwent enucleation of the tumor in November 2020. Pathology showed a well-differentiated neuroendocrine tumor staged T1 pN0 with immunohistochemistry staining diffusely positive for chromogranin, synaptophysin, and insulin with a Ki67 index <3%. In January 2021, at a post-surgery follow-up visit, blood work showed HbA1c 7.1%, proinsulin 31 pmol/L, insulin 13.4 uIU/mL, C-peptide 1.6 ng/mL, BG 193 mg/dl, and chromogranin A 118 ng/mL. HbA1c continued to trend up to 8.0% in January 2022, confirming the resurgence of diabetes, and metformin and pioglitazone were started. Conclusion: Per our literature review, this is the first reported case of a proinsulinoma in a patient with T2DM. In a patient with uncontrolled T2DM, an unexpected amelioration of the disease should raise concern for insulin-secreting tumors, and further workup should be considered. A CGM can be a valuable tool to assess the severity of hypoglycemia in patients with these tumors.

Presentation: Saturday, June 17, 2023
SAT157 Management Of Spontaneous Hypoglycemia In Dialysis-dependent Chronic Kidney Disease Using Diazoxide
Authors: Schledwitz Alyssa , Habbsa Samima Sultana , Villanueva Fiorella Sotomayor , Kim Yoon Kook
Published: October 2023

Disclosure: A. Schledwitz: None. S.S. Habbsa: None. F. Sotomayor Villanueva: None. Y. Kim: None.

Background: Patients with end-stage renal disease (ESRD) are at increased risk of spontaneous hypoglycemia due to decreased renal insulin clearance. Due to its inhibitory effect on pancreatic beta cell insulin secretion, oral diazoxide has been used to treat hypoglycemia due to insulinoma or congenital hyperinsulinemic hypoglycemia, but few cases detail the use of diazoxide in the ESRD population. Here, we present 4 cases of spontaneous hyperinsulinemic hypoglycemia in ESRD successfully treated with diazoxide. Clinical Cases: A 66-year-old man with ESRD presented with altered mental status secondary to severe hypoglycemia despite continuous IV 20% dextrose. Labs revealed glucose 50 mg/dL, insulin 7 (0-3 mcIU/mL), C-peptide 7.8 (0-0.6 ng/mL), proinsulin 35.7 (0-5 pmol/L), beta-hydroxybutyrate [bOH] 0.4 (0-2.7 mg/dL), negative sulfonylurea panel, normal cosyntropin stimulation test and undetectable insulin antibodies, without radiographic evidence of insulinoma. He was started on diazoxide 50 mg TID and remained euglycemic for the remainder of his course. On similar readmission 3 months later, he was restarted on diazoxide with resolution of hypoglycemia within 1 day. A 66-year-old male with ESRD and T1DM treated with pancreas transplant presented with toe gangrene and persistent hypoglycemia. Fasting labs revealed insulin 4, C-peptide 4.5, proinsulin 4.5, and bOH 4.5, without radiographic evidence of insulinoma. He maintained euglycemia 4 days after diazoxide 70 mg BID was started. A 67-year-old male with T2DM and ESRD was admitted for osteomyelitis and 2 years of episodic symptomatic hypoglycemia despite dietary adjustments. Labs showed glucose 59, insulin 7, C-peptide 5.2, proinsulin 7.5, bOH 0.04, and unremarkable imaging. Within 1 day of starting diazoxide 100 mg daily, his blood glucose stabilized, and he was discharged. Finally, a 42-year-old male with ESRD presented with 4 years of persistent episodic hypoglycemia, now refractory to continuous IV dextrose. He was found to have glucose 64, insulin 15, C-peptide 11.5, proinsulin 29, and bOH 0.1, with imaging revealing bulky pancreas without a discrete mass. His hypoglycemic episodes resolved within 1 day of starting diazoxide 100 mg daily. He was readmitted 3 months later for catheter site infection and had no episodes of hypoglycemia with continuation of diazoxide. Conclusion: Diazoxide may be a viable option for the treatment of refractory spontaneous hypoglycemia in patients with dialysis dependent ESRD.

Presentation: Saturday, June 17, 2023
SAT160 Alcoholic Ketoacidosis : An Endocrine Emergency
Authors: Tiu Leo Jr Estabillo , Crisostomo Thelma D
Published: October 2023

Disclosure: L.E. Tiu: None. T.D. Crisostomo: None.

Background: Alcoholic Ketoacidosis is a complex metabolic condition frequently confused with Diabetic Ketoacidosis. Currently, there remains an undefined consensus guideline in the diagnosis and management of this disorder that leads to unreported cases. Clinical Case: A 47-year-old male presented with 2-day history of poor dietary intake due to abdominal pain and vomiting after an alcoholic binge drinking. He is a chronic alcoholic beverage drinker (2 liters of beer per day for 10 years). He has family history of Type 2 Diabetes Mellitus but has no known co-morbidities. Upon admission, vital signs were normal, but he was drowsy and incoherent with signs of dehydration and epigastric tenderness. Initial capillary blood glucose was 52 mg/dL. D50W 50ml intravenously was given with repeat CBG of 222 mg/dL. Arterial blood gas (ABG) showed high anion gap metabolic acidosis (pH 7.0 and anion gap of 42) with undetectable bicarbonate and elevated lactate at 17mmol/L. Serum ketones were elevated at 6mmol/L but Hba1c was normal at 4.6 %. Thus, Alcoholic Ketoacidosis was highly considered. Intravenous hydration with saline solution was instituted then shifted to D10W 125mL/hour to allow insulin infusion therapy at initial rate of 2units/hour. Sodium bicarbonate 100 mEqs was given intravenously with recovery of sensorium thereafter. Oral Vitamin B supplement was started, and two doses of Thiamine 100 mg/IV were given. Other tests showed hyperkalemia (5.28meq/L), hypochloridemia (89.7meq/L), hyperuricemia (12.74mg/dL), and hyperphosphatemia (5.8mg/dL). Serial monitoring and correction of electrolytes were done. Moreover, management integrated simultaneous referrals to other subspecialties to rule out other causes of encephalopathy, to evaluate and manage Hypertriglyceridemic Pancreatitis and Alcoholic Liver Disease, and to facilitate nutritional, psychosocial support and rehabilitation. On 2nd hour, repeat ABG showed improvement of pH to 7.3 and HCO3 to 9.80mmol/L. Lactate decreased to 12.70mmol/L. On the 17th hour, ABG showed metabolic alkalosis with resolution of ketosis hence insulin was discontinued. CBG levels for the next 24-48 hours were within normal limits. Oral feeding was supplemented with parenteral nutrition due to decreased appetite. On 48th hour, he displayed combative behavior with visual hallucinations, but there was no consent for psychiatric referral. On 72nd hour, there was spontaneous marked improvement in his behavior. He was discharged clinically stable. Conclusion: Early recognition of Alcoholic Ketoacidosis is imperative. This is an unrecognized endocrine emergency that portends good outcome with timely diagnosis and management. Treatment includes 1. adequate fluid resuscitation 2. thiamine 3. judicious use of insulin and bicarbonate 4. correction of electrolytes 5. multidisciplinary team management.

Presentation: Saturday, June 17, 2023
SAT162 Management Of Hypoglycemia In Von Gierke's Disease
Authors: Makda Aamir , Sood Aayushi , Syed Omar , Nihan Khuld
Published: October 2023

Disclosure: A. Makda: None. A. Sood: None. O. Syed: None. K. Nihan: None.

Introduction: Glycogen Storage Disease Type-1 (GSD-1)is due to deficiency in Glucose-6-Phosphatase (G6P) results in excessive accumulation of glycogen and fat in the liver, kidney. GSD-1 have a wide spectrum of symptoms including hepatomegaly, hypoglycemia, lactic acidemia, hyperlipidemia, hyperuricemia, and growth retardation. Manifestations of the disease vary in age of onset, rate of disease progression, and severity. Case:18-year-old male with GSD-1a, Von Gierke's disease, hyperuricemia and hypertension presented to the hospital with nausea and vomiting. Patient followed an hourly cornstarch regimen during the day and overnight through infusion via PEG tube. The complaints started at work where he was unable to tolerate oral cornstarch. ABG showed metabolic acidosis, labs showed Leukocytosis, hyperkalemia, acute kidney injury, elevated liver enzymes, hypertriglyceridemia, lactic acidosis. CT abdomen showed hepatomegaly and fatty infiltration with PEG tube intact.

He was admitted to the ICU and started on D5NS for hypoglycemia and lactic acidosis. Per request by the patient's pediatrician, he was transitioned to IV D10- 0.45NS at 110mL/Hr to maintain blood glucose above 75 mg/L. Frequent accuchecks were done till he could tolerate his dietary regimen with cornstarch. Lactic Acid downtrended to 2.9 and accuchecks ranged between 100-110. Cr improved to 1.3, and his home medications (Allopurinol and Lisinopril) were resumed. He was discharged in stable condition with plans for further genetic therapy work up. Discussion: Mainstay therapy for Von Gierke's Disease is prevention of metabolic derangements for which dietary and lifestyle changes are recommended. A low fructose and sucrose diet is recommended with limiting the intake of galactose and lactose to one serving per day. Hypoglycemia treatment in such patients is two fold; utilizing both quick and stable release sources. Cornstarch has been once such therapy since the 1980, its slow digestion provides a steady release of glucose over a longer period of time as compared with other sources of carbohydrates. Dosing guidelines vary from age to age and person to person but it is highly recommended to check BG levels frequently to maintain a BG > 70 mg/dL. Associated high levels of triglycerides and cholesterol can be treated with statins, fibrates, etc. Conclusion: The management of hypoglycemia in GSD 1 disease presents various obstacles which could prove to be fatal. Due to the deficiency of G6P, treatment with a specialized hypoglycemic regimen is warranted. A D10 ½ NS infusion can be used to maintain blood sugar levels as well as correct metabolic or lactate imbalances. Infusion should be gradually weaned off after the patient can tolerate oral feeds as this can help prevent the risk of hypoglycemia and other derangements. Further research is needed in regards to these patients for more sustainable regimens.

Presentation: Saturday, June 17, 2023
OR06-06 Association Between Mixed Dioxin Exposure And Thyroid Functions In U.S. Adults: A Comparison Of Three Statistical Models
Authors: Han Cheng , Lee Sun , Pearce Elizabeth N
Published: October 2023

Disclosure: C. Han: None. S. Lee: None. E.N. Pearce: None.

Background: Previous studies of the relationship between dioxin exposure and thyroid function have usually been based on multi-chemical linear models and results have been inconsistent. However, complex nonlinear relationships and interactions between mixed dioxin exposures and thyroid function have not been studied. We applied three different statistical models to assess the joint effect of 20 dioxins on thyroid function. Methods: A total of 6 polychlorinated dibenzo-p-dioxins (PCDDs), 10 polychlorinated dibenzofurans (PCDFs), 4 dioxin-like polychlorinated biphenyls (DL-PCBs) and serum TSH were measured in the serum of U.S. adults by the National Health and Nutrition Examination Survey (NHANES) from 2007 to 2010. We fitted multivariable logistic regression, weighted quantile sum (WQS) regression, and Bayesian kernel machine regression (BKMR) models to estimate the association of individual and mixed dioxin exposures with high TSH. Results: Among the 2818 individuals included in our analysis, 1.99% had TSH >5.6 uIU/mL. In the multivariable logistic regression established for each single dioxin, 1,2,3,7,8-pncdd, 1,2,3,4,7,8-hxcdd, 1,2,3,6,7,8-hxcdd, 1,2,3,7,8,9-hxcdd, 1,2,3,4,7,8-Hexachlorofuran, 1,2,3,6,7,8-Hexachlorofuran, 3,3',4,4',5,5'-hexachlorobiphenyl, 3,3',4,4',5-Pentachlorobiphenyl, and 2,3,7,8-Tetrachloro-p-dioxin were positively associated with high TSH, and 3,4,4',5-Tetrachlorobiphenyl was negatively associated with high TSH. In the WQS regression model, the WQS index was significantly associated (OR (95% CI): 2.03 (1.06, 3.88)) with high TSH, and 1,2,3,7,8-pncdd (weighted at 0.24) made the largest contribution. In the BKMR analysis, a significant positive association was found between mixed dioxin exposure and high TSH when all dioxins were at their 55 to 70th percentile, compared to their 50th percentile. TSH was lower when all dioxins were at their 30th to 45th percentile, compared to their 50th percentile. Conclusion: By comparing the three statistical models, we concluded that the whole-body burden of 20 dioxins was significantly positively associated with high TSH. 1,2,3,7,8-pncdd played the most important roles in hypothyroidism.

Presentation: Thursday, June 15, 2023
SAT424 Neuroendocrine And Developmental Impacts Of Early Exposure To Estrogenic Endocrine Disruptors
Authors: Streifer Madeline , Mendez Skylar , Raval Raj , Thompson Lindsay , Gore Andrea C
Published: October 2023

Disclosure: M. Streifer: None. S. Mendez: None. R. Raval: None. L. Thompson: None. A.C. Gore: None.

Polychlorinated biphenyls (PCBs) are a class of endocrine disrupting chemical (EDC), exposure to which during early development can result in a variety of neuroendocrine and reproductive abnormalities. Hypothalamic kisspeptin neurons in the anteroventral periventricular nucleus (AVPV) and KNDy (Kisspeptin-Neurokinin B-Dynorphin expressing) neurons in the arcuate nucleus (ARC) are involved in reproductive development, maturation, and success; dysfunctions in kisspeptin signaling can lead to hypogonadotropic hypogonadism and sub- or infertility. KNDy and kisspeptin neurons develop under the influence of gonadal hormones, notably estradiol, and are vulnerable to early exposure to estrogenic EDCs. In this experiment male and female rats were perinatally exposed to the weakly estrogenic PCB mixture Aroclor 1221 (A1221) to determine if and how altered kisspeptin signaling in the ARC and AVPV affects reproductive development. Using RNAscope, a novel in situ hybridization technique, we are exploring if changes in the number of KNDy and kisspeptin neurons or RNA expression of kisspeptin and regulatory peptides neurokinin B and dynorphin are potential mechanisms of the increased incidence of reproductive disorders associated with estrogenic PCB exposure. A1221 exposure did not alter the number of kisspeptin-expressing neurons in the AVPV, though quantification of RNA expression and analysis in the ARC are still ongoing. Serum LH, FSH, estradiol, and testosterone were measured as they are involved in the kisspeptin signaling pathway. In A1221 exposed females, serum LH and FSH were decreased, and estradiol was increased in early life (postnatal day 8). A1221 exposed males had decreased serum LH at a prepubertal time point (postnatal day 30), but FSH and testosterone were unaffected by treatment. Developmental markers that could be indicative of dysfunctions in kisspeptin and KNDy neuron function were monitored. A1221 exposure did not cause any obvious abnormal developmental delays in males or females. Moving forward, we plan on further exploring the effects of developmental A1221 exposure in the AVPV and ARC through gene expression profiling.

Presentation: Saturday, June 17, 2023
SAT427 Effects Of Polychlorinated Biphenyls (PCBs) On Transcription Factor NFκB Activation In Neonatal Rat Microglia Primary Culture
Authors: Dressel Carissa E , Valdez Gia M , Bell Margaret R
Published: October 2023

Disclosure: C.E. Dressel: None. G.M. Valdez: None. M.R. Bell: None.

Polychlorinated biphenyls (PCBs) are a class of persistent organic pollutants that are lipophilic and accumulate in the food chain. Despite bans on PCB production, human exposure continues and can cause disruptions in endocrine, nervous, and immune system functions. As such, PCBs may interfere with the actions of microglia, the innate immune cells of the central nervous system. Healthy microglia are critical for neural development, maintenance of neural functions, and responses to stress, injury, or infection. Initial studies of primary neonatal rat microglial cultures indicate that PCBs alter gene expression of proinflammatory cytokine TNF-α and IL-1β in response to a secondary inflammatory lipopolysaccharide (LPS) challenge. Production of these cytokines is, in part, regulated by transcription factor NFκB. This experiment tests the hypothesis that PCBs alter NFκB activation in neonatal microglia. The forebrains of male and female neonatal Sprague-Dawley rats were dissected to remove meninges, homogenized with collagenase and gentle trituration, and plated on coverslips in DMEM FBS+ media to establish a primary mixed glial culture. After one week, microglia were subcultured using mild trypsin treatment and maintained in conditioned media for 24 hours. The microglia were then exposed to either PCB 153 (0.1 µM) or DMSO vehicle control (0.036%) for 18 hours. After 18 hours, the cells were treated with either LPS (0.5 µg/mL) or PBS vehicle control for 4 hours and then fixed. Immunocytochemistry was used to visualize NFκB and identify microglia, with anti-p65 and Ox42 antibodies, respectively. Coverslips were mounted with DAPI-containing media and the cells were imaged via confocal microscopy. In response to immune activation, NFκB typically translocates from the cytosol to the nucleus. Therefore, we assessed NFκB activation by quantifying p65 fluorescent pixel intensity and percentage of area covered in nuclear or cytoplasmic regions according to DAPI and Ox42 channel masks using ImageJ. A two-way ANOVA was used to identify the effects of PCB vs. DMSO exposure and LPS vs. PBS challenge. Preliminary data indicated that LPS increased the ratio of nuclear to cytosolic NFκB, as expected. PCB exposed cells tended to have lower NFκB presence in the nucleus and cell as a whole, but a greater proportion of NFκB staining localized within the nucleus relative to the rest of the cell. With the current sample (n=4-6 per group, 2-3 males and females each), no interactions between PCBs and LPS were observed. Continued work will increase sample size to explore sex differences and the time course of NFκB response to PCBs, which may help broaden our understanding of the mechanisms by which PCBs affect normal brain activity and disrupt healthy endocrine, immune, and neural development.

Presentation: Saturday, June 17, 2023
SAT431 In-vitro Effects Of Benzophenones 2 And 3 On Cell Proliferation And Autophagy Factors Gene Expression In Lβt2 Cells
Authors: Riaño Gómez Juan Manuel , Sorianello Eleonora , Lux-Lantos Victoria Adela , Webster Nicholas Julian , Fernandez Marina Olga
Published: October 2023

Disclosure: J.M. Riaño Gómez: None. E. Sorianello: None. V.A. Lux-Lantos: None. N.J. Webster: None. M.O. Fernandez: None.

Introduction: Humans and wildlife are exposed to Endocrine Disrupting Chemicals (EDC). Previously, we showed that the in-vitro exposure to Benzophenones 2 and 3 (BP2 and BP3, UV filters) increase cell proliferation in GN11 cells (Susan Wray, USA), and that BP2 and BP3 increase p62 gene expression after 12-h exposure in GT1-7 cells (Pam Mellon, UCSD, USA). In this study, we evaluated the in-vitro effects of BP2 and BP3 on cell proliferation and autophagy factors expression in LβT2 cells (pituitary gonadotropes, Pam Mellon). Materials and Methods: LβT2 cells were plated in 96-well plates (Proliferation Assay) or in 12-well plates (gene expression studies) in DMEM high glucose, with 10% FBS, sodium pyruvate and Penicillin/Streptomicin (complete medium). Before adding the stimuli, media were changed to DMEM without phenol red, with charcoal-stripped FBS (2% for proliferation and 10% for gene expression studies). Proliferation was evaluated using MTS (Promega, WI, USA), in response to BP2 and BP3 (1x10-7 y 1x10-9 M) after 12 or 24-h exposure. Ulk1, Lamp2 and p62 expression was evaluated after 6 or 24-h exposure to BP2 or BP3. RNA was extracted, reverse transcribed and gene expression analyzed by qPCR, using Ppib as control gene. Results were presented as Media±SE and analyzed by ANOVA (Statistica, StatSoft Inc, USA). Results: After 12 or 24-h exposure, BP2 as well as BP3 increased cell proliferation in LβT2 cells [Control-12h:100.0±23.0; BP2-7-12h:201.6±64.0; BP2-9-12h:308.2±80.7; BP3-7-12h:325.2±98.1, BP3-9-12h:339.5±90.4; Control-24h:100.0±24.5; BP2-7-24h: 201.4±45.9; BP2-9-24h:318.2±97.1; BP3-7-24h:289.6±59.9, BP3-9-24h:272.3±99.0; ANOVA: Interaction ns, Main Effect Treatment: BP2-9, BP3-7, BP3-9 different from Control p<0.05, n=5]. After 6 or 24-h exposure, BP3 1x10-9 M increased p62 gene expression, whereas neither BP2 nor BP3 1x10-7 M had an effect on p62 expression [ANOVA: Interaction ns, Main Effect Treatment: BP3-9 different from Control, p<0.05, n=3]. There was no difference in Ulk1 and Lamp2 expression. Conclusions: BP2 and BP3, the EDC evaluated, increased cell proliferation in pituitary gonadotropes, LβT2 cells. Furthermore, BP3 increased expression of p62 at the lower dose used, but not at the higher dose. These results indicate that exposure to these chemicals alter normal physiology in the pituitary, what could promote pituitary pathologies. More results are underway to understand the mechanisms involved. Supported by CONICET, ANPCYT, VA SD Healthcare System, NIH, Fund. Williams, Fund. R. Barón.

Presentation: Saturday, June 17, 2023
SAT432 Interactions Of Endocrine-disrupting Chemical (EDC) Exposure And Fear Recall On Small Noncoding RNA (sncRNA) Contents Of Rat Caput Extracellular Vesicles
Authors: Gillette Ross , Aguirre Meagan , Bell Kiersten , Nadeem Yusra , Thompson Lindsay , Gore Andrea C
Published: October 2023

Disclosure: R. Gillette: None. M. Aguirre: None. K. Bell: None. Y. Nadeem: None. L. Thompson: None. A.C. Gore: None.

Extracellular vesicles (EVs) transfer a complex repertoire of small non-coding RNAs (sncRNA) to sperm as they transit the caput of the epididymis. These molecules are essential for the functional competence of sperm and are directly influenced by environmental factors such as stress, diet, and drug use. We hypothesized that prenatal exposure to EDCs would alter the sncRNA contents of caput extracellular vesicles, and that an exposure to a previously established contextual fear cue would exacerbate this effect. Pregnant dams were fed Nilla Wafers with either 3% DMSO (vehicle control) or Aroclor 1221 (1 mg/kg; a weakly estrogenic EDC) from embryonic day (E)8 until E18 and post-natal day (PND)1 to P21. In adulthood male rat offspring were subjected to a contextual fear conditioning paradigm in which each rat was exposed to an odorant paired with a shock (four 500 ms shocks at 0.7 mA) for three consecutive days. Two weeks later, all rats were returned to the conditioning chambers and exposed to the odorant for 5 minutes (no shocks). The next day, male rats were euthanized and the epididymis was dissected. EVs and total RNA isolated from the caput portion were sequenced on a NovaSeq 6000 SP in a single-end 75 bp format. Reads were checked for quality, trimmed for adapter sequences, aligned to the rat genome, and assigned an annotation designation across 10 different categories of sncRNA. We found that the majority of caput EV sncRNA were either tRNA fragments or piRNA and that unlike mice, miRNA accounted for a small portion of the total reads. The third largest category of aligned reads was in intergenic space and not associated with canonical small non-coding (snc) RNA loci. In-depth investigation determined these reads (∼19 nt) aligned strictly within the boundaries of known CpG islands (CpGi), which have not previously been reported to express any form of sncRNA. Re-exposure to a fear-associated scent 24 hours prior to sample collection substantially reduced the proportion tRNAs in EVs, which were supplanted by an increase in piRNA, CpGi sRNA, and lncRNA. This effect was exacerbated by prenatal EDC exposure and was particularly prevalent in the lncRNA and CpGi sRNA categories. Taken together our data show that the small RNA contents of caput epididymosomes of male rats is substantially different from previous observations reported in mice. We demonstrate that exposure to a contextual fear cue is a powerful influence on sncRNA composition of caput extracellular vesicles and that it is uniquely altered by prenatal EDC exposure. Finally, we provide evidence for what we believe may be a novel category of sncRNA.

Presentation: Saturday, June 17, 2023
SAT433 Improving The Risk Assessment Procedures For Endocrine Disrupting Chemicals Affecting Thyroid Homeostasis: A Scoping Study
Authors: La Motta Concettina
Published: October 2023

Disclosure: C. La Motta: None.

This project aims at improving the risk assessment procedure currently adopted for the endocrine disrupting chemicals (EDCs), focusing in particular on those contained in food contact materials and articles and affecting thyroid homeostasis. EDCs generally display a complex toxicity profile, thus making toxicological evaluation and health risk assessment difficult to achieve. The current standardized methods are rather insufficient to identify and test their complex toxicity, and the application of traditional risk assessment principles may be problematic for this kind of substances. This issue is even more crucial when ECDs are part of food contact materials and articles, which are commonly used by the whole population. The increasingly wider distribution of these products on the global market raised the exposure of consumers to chemical agents on constant and cumulative basis, even for persistent compounds, highlighting the urgency of governing the matter. A number of lists including chemicals exploited for the manufacture of food contact materials and articles have been drawn up by international regulators and public and private bodies. However, besides far from being harmonized, they do not provide systematic information regarding thyroid disrupting properties. Actually, the awareness of science on the thyroid safety profile of compounds intentionally used in the food planet is still limited and fragmented, thus requiring an intensive research activity to fill the existing knowledge gap. We carried out a thoroughly survey of the available databases of chemicals for food contact materials and articles, selecting sixteen EDCs characterized by a high level of uncertainty concerning the toxicological profile and/or dearth of information on human exposure data. The selected compounds should be now investigated by making use of both computational tools and in vitro and in vivo studies, to ascertain their adverse thyroid effects. The resulting outcomes will allow to achieve a reliable risk assessment for the benefit of entire population.

Presentation: Saturday, June 17, 2023
OR01-04 The Growth Hormone Receptor Interacts With Transcriptional Regulator, HMGN1, Upon GH-induced Nuclear Translocation
Authors: Jain Lekha , Vickers Mark H , Jacob Bincy , Middleditch Martin J , Chudakova Daria A , Ganley Austen R D , O'Sullivan Justin M , Perry Jo K
Published: October 2023

Disclosure: L. Jain: None. M.H. Vickers: None. B. Jacob: None. M.J. Middleditch: None. D.A. Chudakova: None. A.R. Ganley: None. J.M. O’Sullivan: None. J.K. Perry: None.

Numerous classical cell-surface receptors that were previously thought to signal exclusively at the cell-surface can also translocate to the nucleus upon ligand binding, with increasing evidence supporting a transcriptional role for examples such as the epidermal growth factor receptor. In addition to classic cell-surface growth hormone receptor (GHR)-mediated signaling, the GHR is rapidly imported into the nucleus of cells upon stimulation with growth hormone (GH). Nuclear GHR localization has been associated with increased cancer cell proliferation. However, whether there are transcriptional changes associated with nuclear GHR import remains to be determined. To investigate whether the GHR interacts with transcription regulators in the nucleus following nuclear translocation we used immunoprecipitation combined with mass spectrometry. GH-dependent phosphorylation of STAT5 was confirmed by western blotting in the human cell-lines RL95-2 (endometrial cancer cell-line), and MCF-10A (mammary epithelial cell-line). The GHR was found to rapidly translocate to the nucleus in both cell lines following GH treatment by immunofluorescence staining, with maximal localisation at 5-10 minutes (p<0.001; RL95-2). Combined immunoprecipitation-mass spectrometry analysis of RL95-2 whole cell lysates following GH stimulation (500 ng/mL, 10 mins) identified 40 novel GHR binding partners, including 3 transcriptional regulators (HMGN1, SUMO1 and DDX21). Gene expression analysis identified 416 genes that were differentially expressed following GH treatment (500 ng/mL, 90 mins). Intersection of differentially expressed genes with known gene targets of HMGN1 (ChIP-Atlas) identified an enrichment of HMGN1 target genes (p<0.001). Co-immunoprecipitation combined with western blot analysis confirmed that HMGN1 associated with the GHR in nuclear extracts. Our results suggest that GHR nuclear translocation might mediate GH actions via interaction with chromatin factors that then drive changes in specific downstream transcriptional programs.

Presentation: Thursday, June 15, 2023
SAT662 Bone Morphogenetic Protein 7 Induces Transdifferentiation Of Human White Preadipocytes Into Thermogenic Beige Adipocytes
Authors: Ali Sahara Leena , Cero Cheryl , Katz Jacqueline , Zhu Alexander , Cypess Aaron Martin
Published: October 2023

Disclosure: S.L. Ali: None. J. Katz: None. A.M. Cypess: None.

Chronic treatment of women with mirabegron, which activates the β3-adrenergic receptor, increases the mass and energy expenditure of thermogenic adipocytes and leads to metabolic benefits including improvement of glucose and lipid homeostasis. In humans, thermogenic adipose tissue is composed of brown and beige/brite (from “brown-in-white”) cells that have different lineages and potentially different functions. Beige cells are of therapeutic interest because they are derived from white adipose tissue, which is present at a considerably higher volume compared to brown adipose tissue. Identifying processes that can transdifferentiate white into beige cells could lead to treatments for obesity-related metabolic disease. We hypothesized that short-term treatment with bone morphogenetic protein 7 (BMP7), a member of the transforming growth factor-β (TGF-β) family of cell regulatory proteins, prior to differentiation could be sufficient to induce white to beige transition in human white preadipocytes. We established paired clonal immortalized human white and brown preadipocyte lines and differentiated the white adipocytes ± BMP7 (3.3 nM) for 7 days prior to differentiation. Cells were collected at d0 (undifferentiated) and d30 (fully mature adipocytes) for gene expression analyses. Short-term exposure to BMP7 in white preadipocytes significantly decreased the expression of white-specific markers TCF21 and HOXC8. The beige-specific markers TBX1 and TMEM26, and the thermogenic marker UCP1 were significantly higher in BMP7-treated white adipocytes. However, long term, 30-day exposure to BMP7 treatment reversed the thermogenic transdifferentiation both human white and brown adipocytes. Basal and drug-induced maximal respiration measured via Seahorse XFe96 Analyzer were significantly higher in short-term BMP7-treated white adipocytes. Our data indicate that BMP7 is capable of transdifferentiating human white preadipocytes into thermogenic beige adipocytes. Given that extended treatment reduced the impact of BMP7 indicates that precise timing of interventions will be needed to utilize white-to-beige transdifferentiation to treat obesity-associated metabolic disease.

Presentation: Saturday, June 17, 2023
THU058 Impact Of A Smartphone App On Follow-up Of Adult Growth Hormone Deficiency (AGHD) Patients: Results From The Management Of AGHD (MAGHD) Study
Authors: De Vincentis Sara , Bellelli Alessio , Golinelli Caterina , Beltrami Elisa , Monzani Maria Laura , Magnani Elisa , Pederzoli Simone , Pacchioni Marco , Orsini Mirko , Diazzi Chiara , Rochira Vincenzo
Published: October 2023

Disclosure: S. De Vincentis: None. A. Bellelli: None. C. Golinelli: None. E. Beltrami: None. M. Monzani: None. E. Magnani: None. S. Pederzoli: None. M. Pacchioni: None. M. Orsini: None. C. Diazzi: None. V. Rochira: Grant Recipient; Self; Pfizer, Inc.

BACKGROUND: AGHD significantly impacts on metabolism, body composition and quality of life (QoL), including sexuality. AIM: To investigate the impact of a smartphone App on AGHD monitoring, especially QoL and comorbidities as well as on both body composition and biochemical parameters. METHODOLOGY: 83 AGHD patients were enrolled in this prospective, open-label, monocentric study (MAGHD Study) of 24-month duration. Patients were evaluated (clinical, biochemical and multidimensional investigations) every 6 months, for a total of 5 visits (from V0, V1, V2, V3 e V4). Multidimensional evaluation included the following questionnaires: QoL-AGHDA, QLS-H, WEMWBS, IPAQ, PSQI e IIEF-15 e FSFI for QoL and sexual function. Body composition was evaluated at V0, V2 and V4 by DEXA. After the first year (Phase 1), patients were grouped according to the acceptance to use (Group 1) or not (Group 2) the smartphone App (MAGHD App) for answering questionnaires. Thus, during the second year (Phase 2), 58 patients provided questionnaires answers through MAGHD App (Group 1) and the remaining 25 patients continued with standard compilation of questionnaires only at visits every 6 months without using MAGHD App (Group 2). RESULTS: The use of the MAGHD App resulted in a higher number of completely filled questionnaires in Group 1 (15.68±8.10) compared to Group 2 (10.33±2.09) (p<0.001). QoL-AGHDA (p=0.001), QLS-H (p=0.030), IPAQ (p<0.001), IIEF-15 (p=0.005) and FSFI (p=0.002) scores were significantly better in Group 1 than Group 2. Compliance to r-hGH therapy increased in Group 1 comparing Phase 1 to Phase 2, but without statistical significance (p>0.05). Comorbidities resulted increased in number in Group 2 compared to Group 1 (p<0,05). No differences were found between Group 1 and 2 in biochemical and hormonal outcomes. CONCLUSIONS: The use of an ad hoc designed smartphone App is well-accepted by patients and associated with a good adherence in its use, suggesting positive effects on QoL in AGHD patients. The integration of a smartphone App in the clinical practice is feasible and may help empowering patients with chronic diseases such as AGHD on daily self-monitoring of their disease trend, beyond helping physicians in tailoring the clinical management. This study belongs to the Pfizer Inc. ‘Independent Grant for Learning and Change, Dissemination & Implementation’ (Grant ID 34515061).

Presentation: Thursday, June 15, 2023
THU059 Increased Intracellular And Extracellular Myocardial Masses On Cardiac Magnetic Resonance Imaging In Patents With Acromegaly
Authors: Wolf Peter , Bouazizi Khaoula , Kachenoura Nadjia , Piedvache Celine , Gallo Antonio , Salenave Sylvie , Maione Luigi , Young Jacques , Prigent Mikaël , Lecoq Anne-Lise , Kuhn Emmanuelle , Agostini Helene , Trabado Severin , Redheuil Alban , Chanson Philippe , Kamenicky Peter
Published: October 2023

Disclosure: P. Wolf: None. K. Bouazizi: None. N. Kachenoura: None. C. Piedvache: None. A. Gallo: None. S. Salenave: None. L. Maione: None. J. Young: None. M. Prigent: None. A. Lecoq: None. E. Kuhn: None. H. Agostini: None. S. Trabado: None. A. Redheuil: None. P. Chanson: Research Investigator; Self; Pfizer. P. Kamenicky: None.

Background: Acromegaly is associated with an increased left ventricular mass, as reported in echo-based and more recently in few cardiac MRI studies. One possible explanation of this increased ventricular mass could be water retention and consequently edema of the ventricular wall. Methods: In this prospective, cross-sectional study 26 patients with active acromegaly and 31 control subjects of comparable age and sex were investigated by cardiac MRI. Patients were explored before and after GH/IGF-I lowering treatment. Cardiac morphology, function and myocardial tissue characteristics were assessed. T2 times were used as a reflect of intramyocardial water content. Results: Ventricular mass (58.08 (54.71; 68.6) vs 46.02 (41.27; 49.82) g/m2; p<0.001) and volume (97.25 (88; 101.21) vs 81.64 (78.08; 96.23) mL/m2; p= 0.0069) were higher in patients compared to controls, without affecting cardiac function. T2 times were not increased in active acromegaly. Both, intracellular (87.9 (71.2; 103.6) vs 67.2 (51.6; 76.9) g/m2; p<0.001) and extracellular (31.9 (26.1; 36.6) vs 21.8 (19.2;24.7) g/m2; p<0.001) myocardial mass were higher in patients compared to controls. GH, but not IGF-I strongly correlated with myocardial mass (r=0.756; p<0.001). In multiple regression analysis, in addition to male sex and HDL cholesterol, the presence of acromegaly was an independent predictor of total myocardial mass and extracellular mass, whereas systolic arterial blood pressure predicted intracellular mass. GH/IGF-I lowering treatment reduced intracellular mass and ventricular volume, without affecting other myocardial tissue characteristics. Discussion: Acromegaly results in a disease specific form of myocardial hypertrophy, characterized by an increase in intra- and extracellular mass, which is reversed after GH/IGF-I lowering treatment. This increase in ventricular mass is thus different to previous observations in essential hypertension. No differences in T2 times suggest against myocardial water retention in active acromegaly as explication of increased extracellular mass.

Presentation: Thursday, June 15, 2023
THU060 Time To First Remission, Medical Therapy And Survival In Acromegaly: UK Acromegaly Register (UKAR) Study
Authors: Deshmukh Harshal , Ssemmondo Emmanuel , Adeleke Kazeem , Batten Lucy , Aye Mo , Sathyapalan Thozhukat
Published: October 2023

Disclosure: H. Deshmukh: None. E. Ssemmondo: None. K. Adeleke: None. L. Batten: None. M. Aye: None. T. Sathyapalan: None.

Introduction: There are limited data on the effect of time to remission of acromegaly and survival in people living with acromegaly Methods: The UK Acromegaly Register collected routine biochemical and clinical data on patients with acromegaly. We defined remission of acromegaly as Growth hormone (GH) controlled at ≤2 μg/l following the diagnosis of acromegaly. Time to first remission was defined as the difference between the date of diagnosis and the first GH date ≤2 μg/l in the longitudinal follow-up data (following surgery and/or medical therapy). We used a t-test to compare the mean survival and used the cox-proportional hazard model to assess the effect of time to remission on survival in acromegaly. Results: The study consisted of 3750 participants in the UK acromegaly register with complete data, of which 2472 (65%) participants achieved remission of acromegaly. The mean time to remission with somatostatin analogs (SA) and dopamine agonist (DA) therapy was 4.5 yrs and 7.3 years, respectively (P<0.001). The mean survival was significantly higher in those who achieved remission of acromegaly as compared to those who did not (25.5 yrs vs 21.9yrs P<0.001) achieve remission. In those who achieved remission, (median follow-up period of 23.9 years), there was 15% mortality in those who achieved remission within 1 year and 23% mortality in those who needed more than 1 year to achieve remission. In the cox-proportional hazard model adjusted for, baseline GH, age at diagnosis, and gender, a shorter time to regression of acromegaly was associated with improved survival (HR=0.92(0.91-0.93) P<0.001). Conclusion: This large population-based study of acromegaly showed that a shorter time to remission is associated with improved survival.

Presentation: Thursday, June 15, 2023
THU089 Natural Course Of Non-Functioning Pituitary Adenoma According To Age And Sex: A Longitudinal Observational Study
Authors: Park Seung Shin , Kang Ho , Ahn Chang Ho , Kim Suhong , Kim Yong Hwy , Kim Jung Hee
Published: October 2023

Disclosure: S. Park: None. H. Kang: None. C. Ahn: None. S. Kim: None. Y. Kim: None. J. Kim: None.

Backgrounds: Non-functional pituitary tumors (NFPTs) are the most common tumor among pituitary adenomas. Surgical treatment is recommended when patients present a mass effect such as a headache, visual field defect, or hormonal dysfunction. However, there is a lack of data regarding the natural course of NFPTs according to sex and age. Herein, we aimed to characterize age and sex-dependent tumor growth in patients with NFPTs. Methods: We enrolled 435 patients (males, n= 162) who were diagnosed with NFPTs and followed up at least one time and more than 12 months from 1997 to 2021 at a tertiary center. Study subjects were divided into three groups according to their age (ages≤50, 51≤age<65, age≥65 years). The characteristics and tumor growth of NFPTs were compared according to sex and age groups. Results: Male patients were older than female patients (median [range], 58.0 [47.0;65.0] vs. 50.0 [39.0;60.0], P<0.001). Annual tumor volume and percent change were higher in male patients compared to female patients (P<0.001, P=0.009, respectively). Despite the shorter follow-up duration in males (61 months in males vs. 71 months in females), male patients underwent surgery more often than female patients (25.9% vs. 11.4%, P<0.001). In male patients, there was no significant difference in the rate of tumor growth between different age groups. However, in female patients, a more rapid increase in annual tumor volume percent change was observed in the older age group compared to the younger age group (2.2 [-3.5;15.2] in females ≤50 years, 6.7 [-2.1;23.7] in 51≤ female <65 years, 10.1 [2.9;25.7] in female≥65 years, P=0.018). The relative risk of tumor growth of more than 20 percent, as compared with the reference group in females younger than 51 years, was 1.81 in the female group aged between 51 to 64 years (95% CI 1.06-3.08, P=0.029), and 2.45 in the female group older than 65 years (95% CI 1.16-5.18, P=0.019) compared to reference group of females younger than 51 years. Conclusion: The NFPT grows more rapidly in males than in females. The growth of NFPT was not significantly different according to age in males, however, in females, the growth of NFPT was more rapid in the older age group. Therefore, males and old females among patients with NFPTs should be more closely followed-up.

Presentation: Thursday, June 15, 2023
SAT669 Development And Biophysical Characterization Of A Humanized FSH-blocking Monoclonal Antibody Therapeutic Formulated At An Ultra-high Concentration
Authors: Rojekar Satish , Pallapati Anusha Rani , Gimenez–Roig Judit , Sant Damini , Gera Sakshi , Korkmaz Funda , Sultana Farhath , Barak Orly , Moldavski Ofer , Cheliadinova Uliana , Gumerova Anisa Azatovna , Miyashita Sari , Kannangara Hasni Suhasha , Frolinger Tal , Witztum Ronit , Macdonald Anne , Georgii Pevnev , Sims Steven Lee , Caminis John N , Meseck Marcia , Ryu Vitaly , Kim Se-Min , Lizneva Daria , Yuen Tony , Zaidi Mone
Published: October 2023

Disclosure: S. Rojekar: None. A.R. Pallapati: None. J. Gimenez–Roig: None. D. Sant: None. S. Gera: None. F. Korkmaz: None. F. Sultana: None. O. Barak: None. O. Moldavski: None. U. Cheliadinova: None. A. Gumerova: None. S. Miyashita: None. H.S. Kannangara: None. T. Frolinger: None. R. Witztum: None. A. Macdonald: None. P. Georgii: None. S.L. Sims: None. J.N. Caminis: None. M. Meseck: None. V. Ryu: None. S. Kim: None. D. Lizneva: None. T. Yuen: None. M. Zaidi: None.

Highly concentrated antibody formulations are oftentimes required for subcutaneous, self-administered biologics. Here, we report the creation of a unique formulation for our first-in-class FSH-blocking humanized antibody, MS-Hu6, which we propose to move to the clinic for osteoporosis, obesity, and Alzheimer’s disease. The studies were carried out using our Good Laboratory Practice (GLP) platform, compliant with the Code of Federal Regulations (Title 21, Part 58). We first used protein thermal shift, size exclusion chromatography, and dynamic light scattering to examine MS-Hu6 concentrations between 1 and 100 mg/mL. We found that thermal, monomeric, and colloidal stability of formulated MS-Hu6 was maintained at a concentration of 100 mg/mL. The addition of the antioxidant L-methionine and chelating agent disodium EDTA improved the formulation's long-term colloidal and thermal stability. Thermal stability was further confirmed by Nano differential scanning calorimetry (DSC). Physiochemical properties of formulated MS-Hu6, including viscosity, turbidity, and clarity, conformed with acceptable industry standards. That the structural integrity of MS-Hu6 in formulation was maintained was proven through Circular Dichroism (CD) and Fourier Transform Infrared (FTIR) spectroscopy. Three rapid freeze-thaw cycles at −80°C/25°C or −80°C/37°C further revealed excellent thermal and colloidal stability. Furthermore, formulated MS-Hu6, particularly its Fab domain, displayed thermal and monomeric storage stability for more than 90 days at 4°C and 25°C. Finally, the unfolding temperature (Tm) for formulated MS-Hu6 increased by >4.80°C upon binding to recombinant FSH, indicating highly specific ligand binding. Overall, we document the feasibility of developing a stable, manufacturable and transportable MS-Hu6 formulation at a ultra-high concentration at industry standards. The study should become a resource for developing biologic formulations in academic medical centers.

Presentation: Saturday, June 17, 2023