bluuebunny/crossref_metadata_2025 huggingface dataset:
MON-010 Effects of Delta-9-Tetrahydrocannabinol (THC) on Oocyte Competence and Early Embryonic Development
Authors: Misner Megan , Favetta Laura A
Published: May 2020

Cannabis is the highest used recreational drug amongst individuals of reproductive age. Fertility clinics advise against cannabis use when undergoing fertility treatments, but the literature backing this statement is weak. This rise in cannabis use has occurred simultaneously with the increase in the percentage of the main psychoactive component of cannabis, delta-9 tetrahydrocannabinol (THC) (1). Current literature suggests that THC mimics the effects of endogenous cannabinoids, binding to cannabinoid receptor 1 (CB1), which has been identified in reproductive tissues (2).

Our research aims to study the impact of THC on oocyte maturation and pre-implantation embryonic development. An in vitro bovine system was used as it is the most appropriate translational model to humans for in vitro reproductive toxicity studies. Bovine oocytes were collected and matured under five treatment groups: control, vehicle (1:1:18 ethanol: TWEEN: saline), low THC (0.032uM), mid THC (0.32uM) and high THC (3.2uM). These doses mimic plasma concentrations reached after therapeutic (0.032uM) or low/high recreational (0.32uM and 3.2uM) cannabis use (3). We hypothesise that THC affects oocyte competence and proper early embryonic development in vitro.

A negative THC dose-dependent response in cleavage rate was observed, with the highest THC group cleaving at 70.2% rate compared to 86.8% and 85.5% of control and vehicle groups, respectively (p<0.0001, n=7). There was no significant difference in blastocyst rate, suggesting that oocyte THC exposure affects the numbers of oocytes capable of development, but those able to cleave will properly reach blastocyst stage. We analyzed changes in gene expression, i) by a full RNA transcriptome analysis (24,128 transcripts screened) and ii) by quantification of Connexin 37 (CX37) and 43 (CX43) mRNA levels. Connexin expression is correlated to oocyte competence (4). RNA transcriptome analysis showed 62 genes that were significantly downregulated only in the low THC group. CX mRNA levels were measured via droplet digital PCR in both cumulus-oocyte complexes (COCs) and blastocysts. No significant differences were detected in blastocysts, however, a significant decrease in both CX37 and CX43 levels was measured in the low THC group in COCs (p<0.05, n= 9). Differences seen exclusively at the low THC dose suggest a role of THC as partial agonist of CB1.

This research aims to understand the effects of cannabis on fertility, as current knowledge during pre-implantation development is limited, making it difficult for physicians to properly advise patients undergoing IVF.

Reference: (1) ElSohly et al., Biol Psychiatry. 2016 Apr 1;79(7):613–9. (2) El-Talatini et al., PLoS ONE. 2009 Feb 24;4(2):e4579. (3) Whan et al., Fertil Steril. 2006 Mar;85(3):653–60. (4) Wang et al., Am J Physiol Endocrinol Metab. 2012 Jun 15;302(12):1511–8
SAT-240 Hypothalamic Lipoma and Growth Hormone Deficiency
Authors: Rochtus Anne Maria , Vinckx Joseph , de Zegher Francis Edouard
Published: May 2020

Introduction: Intracranial lipomas are rare, congenital lesions, most often located at the midline. Most hypothalamic lipomas are asymptomatic, but some cases have been associated with precocious puberty, hypothermia, headache and/or obesity.

Case: A 7-year-old boy was referred for short stature, and proved to be partially growth-hormone deficient. Magnetic resonance imaging (MRI) revealed a lipoma in the paramedian hypothalamus. Growth hormone treatment resulted in swift and uncomplicated catch-up growth.

Discussion: The present case appears to be the first to link hypothalamic lipoma to GH deficiency. The neuro-endocrine pathophysiology underpinning this link remains to be explored.
SAT-252 A Rare Case of Crooke’s Adenoma
Authors: Taiwo Adeyinka , Correia Marcelo , Peyton Nicole
Published: May 2020

BACKGROUND: Crooke’s cell adenoma is a rare variant of ACTH-secreting corticotroph adenomas (<1%). These are aggressive macroadenomas and usually invade the cavernous sinus producing visual symptoms.

Clinical Case: A 48-year old male presented with acute right eye and right temple pain, and diplopia that started 6 days prior. Past medical history included hypertension, type 2 diabetes, hyperlipidemia, traumatic brain injury, CSF leak with bacterial meningitis s/p sphenoid repair and palsy of the right III,

V and VI cranial nerves. Head CT showed a 1.9 x 2.1 x 1.5 cm soft tissue-density mass medial to the right cavernous carotid artery extending into the sphenoid sinus. There was expansile bony change to the floor and right dorsum of the sella. The mass showed mild contrast enhancement and abutted the superior orbital fissure. A brain MRI was not obtained due to history of a left cochlear implant. He did not have any cushingoid features on exam. Neuro-ophthalmology exam revealed, severe sixth cranial nerve palsy, bilateral optic disc pallor and a mild right superotemporal visual field defect. Pre-operative pituitary function tests revealed ACTH 103 pg/ml (7–63), cortisol 14 ug/dl (6.0–18.4), FSH 2.7 mIU/ml (1.5–12.4), LH 3.8 mIU/ml (1.7–8.6), prolactin 17.6 ng/ml (4–15.2), total testosterone 175 ng/dl (249–836), TSH 1.10 uIU/ml (0.27–4.20), free T4 0.67ng/dl (0.8–1.8) and IGF-1 208 ng/ml (69–224). He underwent successful endoscopic trans-sphenoidal resection of a 4.2 x 2.5 x 0.5 cm mass. Pathology revealed hyaline deposits in the cytoplasm and perinuclear cytokeratin in more than 50% of the adrenocorticotroph cells consistent with Crooke’s cell adenoma. Post-operatively, his right eye pain and abduction deficit improved, and diplopia returned to baseline. He was placed on a short hydrocortisone taper. Due to high risk of recurrence, he received 54Gy in 30 fractions to the sella. Six and 12-month follow-up head CTs revealed stable residual enhancing tissue. Pituitary function tests done 9 months post-surgery were normal and no hormone replacement was needed. He was clinically stable 12 months after surgery.

Conclusion: We described a rare case of invasive Crooke’s cell pituitary adenoma with visual disturbances that was managed with resection and radiotherapy. The patient remained free of recurrence 1 year later. However, lifetime surveillance is necessary due to high incidence of recurrence.

Reference:

1.

Crooke’s Cell Tumors of the Pituitary. Neurosurgery, Volume 76, Issue 5, May 2015, Pages 616–22
MON-031 A Gene Expression Profile of the Adolescent Breast and the Impact of Obesity
Authors: Akrobetu Dennis Y , Burkholder Adam B , Ton Thai-Vu T , Kim Susan , Pandiri Arun , Labow Brian I , Nuzzi Laura C , Firriolo Joseph M , Schneider Sallie S , Fenton Suzanne E , Shaw Natalie D
Published: May 2020

Environmental exposures that occur early in life affect breast development and breast cancer (BC) risk in adulthood. Puberty is one such developmental ‘window of susceptibility’ when estrogen (E) stimulates breast adipocytes and stromal and epithelial cells to proliferate at an exponential rate, making them vulnerable to carcinogens. Excess adiposity during adulthood may increase BC risk through obesity-associated inflammation and/or aromatase activity, which increases local E levels. While obesity during puberty might be expected to also increase future BC risk, epidemiological studies suggest that pediatric obesity may actually be protective. The current studies investigated the gene expression profile of the normal adolescent breast and how early life factors such as obesity may influence these profiles.

We performed RNA-seq in 62 histologically-normal breast tissue samples from adolescent girls and young women (mean age 17.8 yrs) who underwent breast reduction surgery. Twenty-nine patients were normal weight (NW; mean BMI 23.2 kg/m2) and 33 were overweight/obese (OB; BMI 31.7). Comparison of our adolescent dataset with published mammary RNAseq datasets from pubertal mice, rats, macaques, and adult women (mean age 38 yrs) revealed relatively poor (~ 30%) overlap with other species, but 88% overlap with adults for the 500 most highly expressed genes in each dataset. The small gene set (n=43) common to all groups was enriched for extracellular matrix components.

We used DESeq2 to identify differentially-expressed (DE) genes in NW vs OB samples. To avoid confounding due to differences in the cellular composition of NW and OB samples, we first used CIBERSORT to computationally estimate the adipocyte fraction of each sample and included this estimate as a covariate. We identified 74 up-regulated and 73 down-regulated genes in NW vs. OB (padj < 0.05). We used Ingenuity Pathway Analysis (IPA) to determine whether the DE genes might reflect activation or inhibition of upstream transcriptional regulators in OB samples. IPA identified the cytokines CSF1 and CSF2 and the chemokine receptor CCR2 as the most highly activated upstream regulators, suggesting a signature of increased inflammation in OB samples. While classical E receptor (ER) targets (e.g., PR, AREG) were not DE’d, IPA identified ESR1, 17-α-ethinyl estradiol, genistein, and PR, as well as growth factors/receptors (EGF, IGF-1, HGF, HER3) and kinases (AKT1, ERK) involved in hormone-independent ER activation, as activated upstream regulators in OB samples.

These studies represent the first investigation of the human breast transcriptome during late puberty and demonstrate that in adolescents, as in adults, OB is associated with increased inflammation which may augment E action in the breast microenvironment.
MON-115 Inappropriate Ordering of Parathyroid Scintigraphy in an Academic Medical Center
Authors: Kimura Brad , Nagelberg Jodi , Koo Sonya , McCowen Karen Clare
Published: May 2020

Introduction: The diagnosis of primary hyperparathyroidism is a biochemical, not radiologic one. Unfortunately, many practitioners even in academic centers order parathyroid scintigraphy to “confirm a diagnosis of adenoma” or distinguish primary from secondary hyperparathyroidism. Knowing the location of single or multiple parathyroid adenomas is unnecessary unless parathyroidectomy is planned. The financial burden of nuclear imaging is substantial. The goals of this study were to determine the proportion of inappropriately ordered parathyroid scans and the cost to the health care system.

Methods: We generated a database of patients who had consulted with at least one physician at our institution and underwent parathyroid scan between December 2012 and December 2017. We focused on the subset that did not undergo parathyroidectomy. “Slicer dicer” software in our EMR was used to generate the database. Chart review extracted data on diagnoses and reasons for parathyroid scintigraphy.

Results: Over 5 years, a total of 325 parathyroid scans were performed. 171 of these did not have parathyroidectomy in our system. However, 18 underwent surgery elsewhere leaving 153 that received parathyroid scans but no surgery (47% of the total). Of the 91 cases so far analyzed of the 153 in our database, average age is 64, with 28 males and 63 females. 61 of the 91 scans (67%) were performed to confirm the diagnosis of parathyroid adenoma; 3 performed because of possible parathyroid adenoma seen on other imaging; and 24 (26%) were done supposedly to localize the adenoma for surgery. Ordering physicians were from primary care (41%), endocrinology (26%), nephrology (18%), and surgery (10%). Final diagnoses for these 91 patients were true primary hyperparathyroidism in 37 (41%), secondary hyperparathyroidism in 38 (42%), unclear in 10 and FHH in 4. In the primary hyperparathyroidism group, 19/37 met criteria for consideration of parathyroidectomy, but only 5/19 received surgical consultation. These 5 patients either refused surgery or surgeon decided against, usually because of high surgical risk.

Conclusion: 47% of parathyroid scans at an academic institution were performed in patients who did not undergo parathyroidectomy. Many parathyroid scans were ordered inappropriately to “confirm” a diagnosis of primary hyperparathyroidism, leading to unnecessary charges and resource waste. Physician charges for sestamibi scans range from $237-$1942, depending on whether planar imaging, SPECT, or SPECT-CT is used; hospital charges are $1165-$3211. We propose to change the ordering system for parathyroid imaging to clarify that this is not a method to diagnose parathyroid adenoma, rather a tool to optimize surgical planning when the diagnosis is secure.
MON-040 11-Oxygenated C19 Steroids in Polycystic Ovarian Syndrome
Authors: Mody Armaiti , Lodish Maya Beth , Auchus Richard Joseph , Huddleston Heather Gibson
Published: May 2020

BACKGROUND: Polycystic ovarian syndrome (PCOS), an endocrine and reproductive disorder consisting of hyperandrogenism, menstrual dysfunction and ovarian changes, affects 6–20% of reproductive aged women worldwide. While hyperandrogenemia is traditionally determined by evidence of elevated testosterone (T), this hormone can be difficult to accurately measure in women with relatively lower circulating levels compared to men. Recent studies have suggested that four adrenal androgens known as 11-oxygenated C19 steroids (11OxyAs), specifically 11-ketotestosterone (11KT), may be good alternative markers for hyperandrogenism in PCOS. Using a multiethnic population seeking evaluation for PCOS symptomatology, we sought (1) to investigate the utility of 11OxyAs to differentiate women with and without NIH PCOS relative to classical androgens such as T, androstenedione (A4) and DHEAS levels, and (2) to evaluate the relationship of 11OxyAs to clinical findings of androgen excess.

Methods: Using the University of California, San Francisco PCOS Tissue Bank, serum samples from 131 women seen for a PCOS evaluation were selected sequentially and identified as PCOS or non-PCOS (controls) based on meeting NIH criteria at the time of evaluation. In addition to obtaining gonadotropin and metabolic profiles, classical androgens and 11OxyAs were measured using mass spectrometry. The relationship of these androgens to modified Ferriman-Gallwey (mFG) scores and ovarian morphology were also assessed.

Results: Out of 131 women selected, 83 met NIH PCOS criteria at the time of evaluation and 48 did not (controls). Age and BMI did not differ among the two groups. As expected, total T, A4 and LH were all significantly higher in NIH PCOS. A trend towards higher HOMA-IR levels was also seen in NIH PCOS, but this did not reach statistical significance (3±3.9 mg/dL vs. 1.9±1.7 mg/dL, p = 0.12). No difference was seen in all four 11OxyAs between NIH PCOS and controls. Unlike previous studies, we also did not find mean 11KT levels to exceed that of T in both controls (T 393±143 pg/mL vs. 11KT 389±206 pg/mL) and PCOS (T 530±245 pg/mL vs. 11KT 388±201 pg/mL). In addition, no relationship was seen between HOMA-IR and 11β-hydroxyandrostenedione (11OHA4) or 11-ketoandrostenedione (11KA4) levels. Within PCOS, DHEAS and A4 were noted to have a weak but inverse relationship to BMI (r2 0.05 p = 0.05; r2 0.08 p = 0.007), whereas no correlation was seen between any of the four 11OxyAs or T and BMI. Lastly, 11OxyAs, T, and A4 levels did not predict mFG scores or polycystic ovarian morphology.

Conclusions: 11OxyAs levels were not statistically higher among women with NIH PCOS compared to at risk women who did not meet NIH criteria. There was no significant relationship between these androgens and mFG scores or ovarian morphology. Further studies are necessary to show the utility of 11OxyAs levels as a marker for hyperandrogenism or metabolic risk.
MON-613 The Expression of TBC1 Domain Family, Member 4 (TBC1D4) in Skeletal Muscles of Insulin-Resistant Mice in Response to Sulforaphane
Authors: Rizk Nasser M , Saleh Amina , ElGamal Abdelrahman , Elsayegh Dina , Cakir Isin , Ghamari-Langroudi Masoud
Published: May 2020

The Expression of TBC1 Domain Family, member 4 (TBC1D4) in Skeletal Muscles of Insulin-Resistant Mice in Response to Sulforaphane.

Background: Obesity is commonly accompanied by impaired glucose homeostasis. Decreased glucose transport to the peripheral tissues, mainly skeletal muscle, leads to reduced total glucose disposal and hyperglycemia. TBC1D4 gene is involved in the trafficking of GLUT4 to the outer cell membrane in skeletal muscle. Sulforaphane (SFN) has been suggested as a new potential anti-diabetic compound acting by reducing blood glucose levels through mechanisms not fully understood (1). The aim of this study is to investigate the effects SFN on TBC1D4 and GLUT4 gene expression in skeletal muscles of DIO mice, in order to elucidate the mechanism(s) through which SFN improves glucose homeostasis.

Methodology: C57BL/6 mice (n=20) were fed with a high fat diet (60%) for 16 weeks to generate diet induced obese (DIO) mice with body weights between 45–50 gm. Thereafter, DIO mice received either SFN (5mg/kg BW) (n=10) or vehicle (n=10) as controls daily by intraperitoneal injections for four weeks. Glucose tolerance test (1g/kg BW, IP) and insulin sensitivity test (ITT) were conducted (1 IU insulin/ g BW, IP route) at the beginning and end of the third week of the injection.

At the end of 4 weeks of the injection, samples of blood and skeletal muscles of both hindlimbs were collected. The expression levels of GLUT4 and TBC1D4 genes were analyzed by qRT-PCR. Blood was also used for glucose, adiponectin and insulin measurements.

Results: SFN-treated DIO mice had significantly lower non-fasting blood glucose levels than vehicle-treated mice (194.16 ± 14.12 vs. 147.44 ± 20.31 mg/dL, vehicle vs. SFN, p value=0.0003). Furthermore, GTT results indicate that the blood glucose levels at 120 minutes after glucose infusion in was (199.83±34.53 mg/dl vs. 138.55±221.78 mg/dl) for vehicle vs. SFN with p=0.0011 respectively. ITT showed that SFN treatment did not enhance insulin sensitivity in DIO mice. Additionally, SFN treatment did not significantly change the expression of TBC1D4, and GLUT4 genes in skeletal muscles compared to vehicle treatment (p values >0.05).

Furthermore, SFN treatment did not significantly affect the systemic insulin (1.84±0.74 vs 1.54±0.55 ng/ml, p=0.436), or adiponectin (11.96 ±2.29 vs 14.4±3.33 ug/ml, p=0.551) levels in SFN vs. vehicle-treated DIO mice, respectively.

Conclusion: SFN treatment improves glucose disposal in DIO mice, which is not linked to the gene expression of GLUT4 and TBC1D4 and its mechanism of glucose disposal in skeletal muscles. Furthermore, SFN treatment did not improve insulin level, and the insulin sensitizer hormone adiponectin as potential players for enhancing insulin sensitivity.

1.

Axelsson AS, Tubbs E, Mecham B, Chacko S, Nenonen HA, Tang Y, et al. Sci Transl Med. 2017;9(394).
SUN-908 Mediastinal Paraganglioma: A Rare Presentation of a Rare Tumor
Authors: Deepika Fnu , Andre Karla Bermudez Saint , Kansara Abhishek
Published: May 2020

BACKGROUND

Paragangliomas (PPGL’s) are extra-adrenal chromaffin-cell originating tumors that arise from the para-aortic ganglia and are exceptionally rare making up only 1% of the mediastinal tumors. They are slowly growing and highly vascular tumors associated with high morbidity and mortality due to proximity and invasion into the heart, great vessels. The majority of them are asymptomatic and hence discovered incidentally. Surgical resection remains the standard of care with good long term survival. We describe a case of a young male with middle mediastinal PPGL who remained asymptomatic despite large functional tumor.

CLINICAL CASE

A 38-year old Arab male with no previous smoking history was referred to our hospital for evaluation of mediastinal mass that was incidentally noted on an echocardiogram done 1 year ago in his home country when he presented with intermittent exertional shortness of breath for 10 years. The workup done there with CT chest revealed a large mediastinal mass measuring 10.4×8.8×8.6 cm extending into ascending aorta, right pulmonary artery, right superior pulmonary vein, right and left atria as well as the main bronchi bilaterally invading the superior vena cava. However, he had no further treatment until he was seen 1 year later in the US at another institution where biochemical testing showed elevated plasma norepinephrine of 2697 (70-750 pg/ml, supine), dopamine 7667 (<30 pg/ml). 24hr-urine studies showed elevated normetanephrine of 3418 (111-419 mcg/24hr), metanephrine 3488 (200-614 mcg/24hr), and Homovanillic acid (HVA) 51.5 (<0.8 mg/24hr). He underwent endobronchial ultrasound with mediastinal tissue biopsy of the mass that revealed pheochromocytoma. The patient was subsequently referred for cardiovascular intervention at our hospital and endocrinology was consulted for pre-operative preparation. One week before surgery, he was started on doxazosin 2mg daily followed 3 days later by propranolol 10 mg twice daily and recommend high salt and fluid intake. The patient underwent surgical resection of the mass and pathology showed mediastinal PPGL. The postoperative course was complicated by bradycardia with the placement of a permanent pacemaker. He recovered well after surgery with the resolution of symptoms and a significant decline in catecholamines to urine normetanephrine of 385 mcg/24hr and metanephrine 497 mcg/24hrs. There was no evidence of metastatic or residual disease on follow-up CT chest 5 months later.

CONCLUSION

There are limited case reports of PPGL in the middle mediastinum. This case highlights that mediastinal PPGL’s can remain clinically silent for many years prior to the presentation which can lead to delayed diagnosis, yet a strong collaborative team approach between oncology, cardiovascular surgery and endocrinology can confer favorable clinical outcome.
SUN-489 MetastaticThyroidCancer
Authors: Vásquez Romyna La Rosa
Published: May 2020

Background In many cases, thyroid cancer leads to metastasis; however, isolated metastasis to the liver from thyroid cancer is a rare event. When this occurs, patients have a very poor prognosis. Case Study A clinical case of a 33-year-old woman, born in Piura and coming from Lima, with no major history, diagnosed with a 4-month-old cervical tumor is presented. An ultrasound scan and a further biopsy were performed before an eventual diagnosis of papillary thyroid carcinoma. Operation was performed, and a classical papillary carcinoma of 0.90 inches was detected, along with macro-metastasis in a parathyroid ganglion. Post-surgical thyroglobulin was 1071 ng/ml (n< 50 ng/ml). The tomography in her lung showed three nodules in the middle lobe. A further dose of 150 millicuries of radioactive iodine (I-131) was given, with whole-body scanning, post positive in both lung fields and right lank pain. The tomography in the abdomen revealed a hepatic pedicle injury, compatible with teratoma. A liver resection surgery was scheduled, and metastatic papillary carcinoma was identified. A V600 mutation in BRAF gene was present in thyroid gland and not detectable in the liver. Conclusions This case shows an example of thyroid cancer with uncommon metastasis in the liver, which occurs in 0.5% of all thyroid metastases. It is even rarer that positive iodine was found. Liver metastasis represents a poor prognosis, however it has been reported that resective surgery offers patients a better chance of survival. Multiple factors influence its pathogenesis, including BRAF mutations. In this case, mutation was detected in thyroid gland, but not in liver metastasis, which could represent diverse BRAF mutations.
SAT-471 A Rare Case of Pretibial Myxedema Preceding Graves’ Hyperthyroidism
Authors: Bao Ginny W , Kirkeby Kjersti Meyer , Kwan Christina
Published: May 2020

Introduction:

Pretibial myxedema, also known as thyroid dermopathy, is typically seen as a manifestation of long-standing Graves’ disease. We report a rare case of pretibial myxedema as the initial presentation of Graves’ disease in a patient without ocular or overt hyperthyroid symptoms.

Case:

The patient is a 68 year old male with a past medical history of type 2 diabetes and hypertrophic cardiomyopathy, presenting with a one year history of a mildly pruritic, violaceous plaque of his left shin. He was referred to Dermatology Clinic, and biopsy came back as pretibial myxedema. Of note, his thyroid function studies were normal 6 months prior, and he had no symptoms of active thyroid disease at presentation. Additionally, he did not have any ocular abnormalities on exam. Upon repeat laboratory studies, he had low but detectable TSH, normal free T4, mildly elevated free T3, and elevated TSI/TRAb, overall consistent with early Graves’ hyperthyroidism. His pretibial myxedema was treated with high-dose topical steroids with improvement of his dermopathy. For his hyperthyroidism, he was started on low dose methimazole and never developed Graves’ ophthalmopathy.

Discussion:

Although the exact pathophysiology of pretibial myxedema is unknown, it is thought to be mediated by TSH receptor expression on fibroblasts, similar to Graves’ ophthalmopathy. It is usually seen in patients with active or longstanding Graves’ disease, often with high levels of thyroid receptor antibodies and concomitant Graves’ ophthalmopathy. However, there are a few cases of pretibial myxedema occurring in euthyroid patients with or without ocular symptoms, usually in the setting of elevated TSI, TRAb, or TPO antibodies. Our patient is a rare case of pretibial myxedema preceding active thyroid disease without ophthalmologic manifestations. Patients with hyper-pigmented, non-pitting plaques or nodules on pretibial regions should prompt providers to consider pretibial myxedema and test for thyroid studies, even in those without symptoms of hyperthyroidism or Graves’ ophthalmopathy.
SAT-049 Testosterone Therapy Reduces Inflammatory Activation of Human Monocytes in Hypogonadal Type-2 Diabetic Men as a Potential Mechanism to Improve Atherosclerosis
Authors: Bateman Lauren , Rao Preethi , Jones Thomas Hugh , Kelly Daniel Marcus
Published: May 2020

Testosterone deficiency is prevalent in men with type 2 diabetes (T2D) and is associated with greatly elevated risk of cardiovascular mortality. Testosterone replacement has beneficial effects on surrogate markers and risk factors of atherosclerosis including inflammation, cholesterol and insulin resistance improving survival in men with T2D. The underlying mechanisms of this action remain poorly understood. Inflammation is a central feature to both T2D and atherosclerosis and is driven by monocyte/macrophages, placing these immune cells at the crossroads of disease pathology. The recruitment of immune cells to atherosclerosis-prone areas of the vasculature is influenced by many factors including the inflammatory status of the circulating monocytes. The present study investigates the influence of testosterone replacement on monocyte inflammatory markers in a randomised double blinded placebo controlled clinical trial. 65 men with poorly controlled diabetes (HbA1c between 53 and 80 mmol/mol) and confirmed hypogonadism via early morning [0800−1200h] total testosterone ≤12 nmol/L or calculated free testosterone ≤255 pmol/L on two occasions ≥1 week apart, with at least two symptoms of hypogonadism were included in the study. Patients were randomly assigned to either placebo or treatment (depot testosterone undecanoate, 6 weekly followed by 3 monthly intramuscular injections) for 6 months. Monocytes were isolated from whole blood collected at baseline, 3 and 6 month visits followed by gene expression of key inflammatory targets IL-1β, IL-6, IL-10, TNFα, ICAM1, TLR2, TLR4, SCARB1 and MCP1 assessed via qPCR. Pro-inflammatory targets TNFα and MCP1 were significantly reduced over time in monocytes from patients treated with testosterone between 3 and 6 months (1.39±0.39 Vs 0.68±0.09, P<0.01; 15.36±7.79 Vs 1.88±0.93, P<0.01 respectively) and TNFα at 6 months compared to the start of the study (1.00±0.00 Vs 0.68±0.09, P<0.001) when normalised to baseline. TNFα expression was also significantly reduced compared to placebo treated patient monocytes at 6 months (0.68±0.09 Vs 3.45±1.50, P<0.01). Other targets were not significantly altered over time or between treatment groups. These findings importantly indicate for the first time that testosterone influences monocyte inflammatory activation in type 2 diabetic men by altering expression of the most potent atherogenic chemokine MCP1 and potent pro-inflammatory cytokine TNFα, as a potential mechanism to protect against atherosclerotic plaque development in hypogonadism.
MON-727 The Chromatin Landscape of Glucocorticoid Regulated Genes in Mouse Embryonic Neural Stem / Progenitor Cells
Authors: Berry Kimberly , Chandran Uma , Mu Fangping , DeFranco Donald
Published: May 2020

Stem/Progenitor Cells

Antenatal administration of Dexamethasone (Dex), a synthetic glucocorticoid (GC), is a common clinical intervention for women at risk for preterm birth or in preterm labor that effectively reduces fetal risk of mortality and bronchopulmonary-related comorbidities. Despite the therapeutic potential of Dex, excess GC act adversely in the developing central nervous system to reprogram distinct neural circuits in the brain by acting through the glucocorticoid receptor (GR). For example, prenatal exposure to excess GCs can impact neural stem and progenitor cell (NSPC) proliferation leading to long-term alterations in prefrontal cortical neuronal complexity, which could contribute to behavioral and cognitive impairments later in life. The GR is a member of the nuclear receptor superfamily that, when bound by a ligand, translocates from the cytoplasm to the nucleus and associates indirectly or directly with DNA elements (e.g. glucocorticoid responsive elements or GREs) resulting in the activation and/or repression of target genes. While GR-regulated transcriptomes have been identified in many NSPC models, the mechanisms responsible for programming these cells for GC-responsiveness remain largely unknown. We therefore used transposase accessible chromatin followed by genome-wide sequencing (Omni ATAC-seq) to characterize the chromatin landscape of primary embryonic mouse NSPCs in response to an acute in vitro treatment with Dex. We identified a small, yet distinct fraction (0.002%, p<0.05) of open chromatin sites that were Dex-inducible. 95% of these Dex-induced changes in chromatin accessibility occur within intronic or intergenic regions, suggesting the presence of long-range enhancer-promoter contacts that mediate NSPC transcriptional responses to Dex. Motif enrichment analysis revealed putative GRE sites located in Dex-inducible open chromatin within -5kb/+2kb of a Dex-induced gene, providing possible DNA targets of GR for further validation. A number of other transcription factors implicated in neurodevelopmental processes were found to underlie both Dex-inducible and constitutively open chromatin regions. Characterization of the precise epigenetic and transcriptional response to excess GC in-utero, and its influence on acute and chronic neurological outcomes, will encourage the development of alternative GC treatment regimens that could protect the developing brain from insult while providing optimal health outcomes in neonates.
SUN-679 Early Onset Diabetes Mellitus in a Young Woman with Ring Chromosome 13 Syndrome
Authors: Buehler Lauren Anne , Mikhael Alexandra , Zimmerman Robert S
Published: May 2020

Background: Ring chromosome 13 is a rare genetic disorder in which the two ends of chromosome 13 are fused together to form a ring shape. Ring formation subsequently leads to gene deletions on both the long and short arms of the chromosome. Observed phenotypes among these patients are variable due to differences in the number of genetic deletions that occur with fusion. Common clinical features seen in ring chromosome 13 include micro- or anencephaly, severe mental retardation (MR), ambiguous genitalia, growth retardation, and facial dysmorphism. To our knowledge, diabetes mellitus (DM) has only been reported in two published case reports of patients with ring chromosome 13. It has been proposed that development of DM in this syndrome may be due to deletion of the IRS2 gene on the long arm of chromosome 13. This hypothesis is based on evidence from knockout mice studies and genetic comparisons of ring chromosome 13 patients with and without DM.

Clinical Case: We present a case of a 23-year-old woman with a known history of mosaic ring chromosome 13 abnormality who was diagnosed with DM at age 21. Diagnosis was made incidentally based on laboratory data obtained at a routine outpatient visit. Hemoglobin A1c at that time was 13.1%. Glutamic acid decarboxylase and insulin antibodies were negative. She has been on a basal-bolus insulin regimen since diagnosis with overall good glycemic control (average hemoglobin A1c 6.5%). She has no known micro- or macrovascular complications of DM. She has no family history of chromosome 13 syndrome or DM. Other notable clinical features associated with her ring chromosome 13 disorder include MR, spastic encephalopathy, delayed puberty (menarche at age 18), scoliosis, facial dysmorphism, and deficiencies in clotting factors 7 and 10.

Conclusion: DM is a common disorder that is typically multifactorial in etiology. In rare cases, DM can be the result of monogenic mutations or deletions. We present a case of a young woman with early-onset antibody-negative DM thought to be related to gene deletions resulting from her underlying ring chromosome 13 abnormality. There is some evidence from prior case reports and rodent studies to suggest that DM in patients with this disorder may be the result of IRS2 deletion from the long arm of chromosome 13. Although DM is a rare complication of ring chromosome 13 disorder, early screening should be considered in these patients to prevent development of downstream complications.

References:

1. Babaya N, Noso S, Hiromine Y, Ito H, Taketomo Y, Yamamoto T, Kawabata Y, Ikegami H. Early-Onset Diabetes Mellitus in a Patient With a Chromosome 13q34qter Microdeletion Including IRS2. J Endocr Soc. 2018 Sep 11;2(10):1207-1213.

2. Lagergren M, Börjeson M, Mitelman F. Prophase analysis of ring chromosome 13--an attempt at phenotype-karyotype correlation. Hereditas. 1980;93(2):231-3.
MON-625 Utility of Continuous Glucose Monitoring in Children with Type 1 Diabetes: Is HbA1c Enough?
Authors: Owens Lindsey L , Chalise Sweta , Vyas Neha , Gurnurkar Shilpa
Published: May 2020

Introduction: Type 1 diabetes is an autoimmune condition resulting in insulin deficiency that requires daily insulin therapy and self-monitoring of blood glucose. Continuous glucose monitoring (CGM) systems allow for measurement of interstitial fluid glucose levels in a continuous fashion to identify variations and trends that are not feasible with conventional self-monitoring. Hemoglobin A1C (HbA1C) is the method used to assess adequate glycemic control and relates to future risk of developing complications. Current evidence has shown improvement in HbA1C with concomitant use of CGM in adults over 25 years of age with Type 1 diabetes, whereas studies in children and adolescents have failed to show this. However, it is important to note the limitations in HbA1C use as it is a marker of average blood glucose over 3 months but does not reflect glycemic variability. More recent data has suggested that factors such as time in range (TIR), which can be determined with CGM use, are also associated with decrease risk of diabetes complications.

Methods: The goal of our study was to analyze the change in HbA1C levels after using a CGM (DEXCOM G4, G5, G6) over a 6-month period in pediatric patients with Type I diabetes. Two HBA1c levels 3 months apart from 92 patients were collected before using a CGM and two while using a CGM. Results were compared by using a dependent samples t-test. IBM SPSS 25.0 was used for data analysis.

Results: Preliminary analysis indicates the average change in HBA1C among the patients (N=92) before (-0.08 ± 1.16) and while using the CGM (0.12 ± 1.00) was not significantly different (t (79) = -1.27, p = 0.21). The average change in HBA1C was also not significantly different (p>0.05) among the patients before and while using the CGM for gender (males and females), age groups (0-7 years, 8-14 years, and 15-24 years), and generations of DEXCOM used (G4, G5, and G6).

Conclusion: As has been shown in other studies, we did not find a significant change in HbA1c after CGM use for 6 months in our patients. While HbA1C is a reflection of blood sugars over a 3-month period, it does not provide information about glycemic excursions. Metrics derived from CGM use, such as TIR, can provide actionable information which we did not address in our study. There have been reports of the association between TIR and long-term complications of diabetes. Most data comes from studies in adults and pediatric data is lacking. We propose that future studies must look into CGM metrics such as TIR to better define glycemic control in pediatric patients with diabetes mellitus.
OR05-05 Lethal ERα-Dependent Hyperactivation of the Unfolded Protein Response Induces Complete Regression Without Recurrence of Primary and Metastatic Breast Cancer
Authors: Duraki Darjan , Boudreau Matthew W , Wang Lawrence , Mao Chengjian , Tang Bingtao , Ma Liqian , Roy Edward J , Fan Timothy M , Park Ben Ho , Nelson Erik R , Hergenrother Paul J , Shapiro David J
Published: May 2020

Metastatic estrogen receptor α (ERα) positive breast cancer is presently incurable and most patients die within 7 years. From a medicinal chemistry program, we identified a novel small molecule that acts through ERα to kill breast cancer cells and often induces complete regression without recurrence of large, therapy-resistant primary breast tumors and of lung, bone, and liver metastases. To target metastatic ERα positive breast cancer, we exploited our finding that estrogen-ERα activates an extranuclear tumor-protective, signaling pathway, the anticipatory unfolded protein response (UPR). We repurposed this tumor protective pathway by targeting it with the small molecule, ErSO. ErSO kills cancer cells by acting non-competitively through ERα to induce lethal hyperactivation of the anticipatory UPR, triggering rapid necrotic cell death. Using luciferase to image primary tumors and metastases containing lethal ERαD538G and ERαY537S mutations seen in metastatic breast cancer, oral and injected ErSO exhibited unprecedented antitumor activity. In mouse xenografts bearing large breast tumors, oral and injected ErSO induced complete regression (>115,000 fold mean regression) in about 45% of mice (18/39). Although durable response for 4-6 months without additional treatment was common, tumors that did recur remained fully sensitive to ErSO re-treatment. Consistent with the essential nature of the UPR pathway targeted by ErSO, in more than 100 tumor-bearing mice, we have never seen an ErSO-resistant tumor. In just 7 days, oral ErSO induced complete regression of most lung, bone, and liver metastases. ErSO is well-tolerated in mice and blood-brain-barrier penetrant. Injected ErSO induced profound regression of challenging brain tumors. On average, ErSO-treated tumors were >180-fold smaller than vehicle-treated tumors. These xenograft studies used human cancer cells in mice that lack a functional immune system and therefore did not exploit the known ability of inducers of necrotic cell death to activate immune cells and induce immunogenic cell death. Notably, medium from breast cancer cells killed by ErSO contained high levels of immune cell activators, robustly activated mouse and human macrophages and increased macrophage migration. Moreover, use of ErSO is not limited to breast cancer. ErSO rapidly kills ERα positive ovarian and endometrial cancer cells that do not require estrogen for growth. ErSO’s potent activity against advanced primary and metastatic ERα-positive breast cancers represents a paradigm shift in leveraging ERα for anticancer efficacy.
SAT-013 Segesterone Acetate (SA) Serum Levels with a Statistical Model of Continued Use of the SA/Ethinyl Estradiol (EE) Contraceptive Vaginal System
Authors: Liu James H , Archer David F , Simon James A , Kaunitz Andrew M , Plagianos Marlena , Bernick Brian , Mirkin Sebastian
Published: May 2020

The novel, ring-shaped, contraceptive vaginal system (CVS; Annovera™), contains 103 mg of SA and 17.4 mg of EE, and delivers a mean dose of SA 0.15/EE 0.013 mg/day. The CVS was designed to last thirteen cycles in a 21 day-in/7 day-out regimen. Objectives of these analyses were to determine the amount of SA/EE remaining in the CVS after 13 cycles of use and to estimate the SA level in serum after 1 year of continuous use.

Residual SA/EE levels from CVS used by women (n=18) for 13 cycles of 21/7 regimen in a phase 3 clinical trial were further analyzed in vitro. Hypothetical SA level in serum after 1 year of continuous use was estimated using data from 39 women who participated in a 13-cycle pharmacokinetic study of the 21/7 CVS regimen. The serum data were used to construct a statistical model for the change in serum SA concentrations from cycle 1 to cycle 13. The data from each individual subject was modeled to estimate the serum SA level based on days of use. Each subject’s model was then used to estimate the serum SA level after 364 days of continuous CVS use. The average of all 364-day values from valid models was then calculated.

After 13 cycles of a 21/7 regimen, the amount of SA/EE remaining in the CVS was 61.7 mg/14.0 mg (60%/80% of the original SA/EE load), which established the release of 0.15 mg SA and 0.013 mg EE per day for a total of 273 days. The model predicted that the mean serum SA level after 364 days of continuous use was 71 pmol/L with a lower limit of the 90% CI of 52 pmol/L.

A high percentage of SA/EE remained in the CVS after 13 cycles of use. Serum SA levels predicted by the model following 1 year of continuous use were similar to those previously observed (mean 73 pmol/L) at 1 year in a SA silicone implant trial in which there were no reported pregnancies at 1 year (Sivin et al, Contraception. 2004;69:137-144). No pregnancy or PK data with continuous use of the CVS is available at this time. Further study on the continuous use of the CVS is warranted.
SUN-124 Are Venous Thromboembolic Events Increased in MEN1 Patients?
Authors: Lee Maya E , Sustache Yashira M Ortega , Agarwal Sunita Kishore , Tepede Aisha , Mandl Adel , Bansal Rashika , Tirosh Amit , Piaggi Paolo , Weinstein Lee Scott , Simonds William F , Blau Jenny
Published: May 2020

Background: Multiple endocrine neoplasia type 1 (MEN1) is a rare inherited disorder in which patients develop multiple simultaneous hormone-secreting tumors. Most common tumors include: anterior pituitary adenomas (50%), multi-gland parathyroid adenomas (95%), and gastroenteropancreatic neuroendocrine tumors (40-80%). Only rare MEN1 associated glucagonomas (<1%), and ACTH-producing neuroendocrine tumors (<5%) are known to increase risk of hypercoagulability. It is unknown if patients with MEN1 syndrome have increased risk of venous thrombolytic events (VTE), defined as a deep-vein thrombosis and/or pulmonary embolism.

Methods: We queried a prospective natural history study of MEN1 patients who tested positive for germline MEN1 mutations (n=287) between 1991-2019 (54 patients on our current protocol were followed before 1991; the earliest was 1971). All lifetime events of VTE were included. Search terms included: DVT, thromb, embol, PE, pulmonary embolism, clot, hematology consult, anticoagulant, coumadin, lovenox, xarelto, warfarin, aspirin, rivaroxaban and apixaban. After initial screening, 10 patients were removed due to insufficient clinical data. Kaplan-Meier analysis was performed to compare age of death between the two cohorts. Results were expressed as mean ± standard deviation.

Results: Thirty-four subjects (mean 57.5 years-old, 17 women) were identified with any VTE, yielding a prevalence rate of 13.4%. The incidence of VTE corresponded to 264 events per 100,000 patient-years, which was ~2-fold higher than the estimated annual incidence rate in the general population (104-183/100,000 patient years).1 Kaplan-Meier analysis revealed no significant difference in survival between the two groups (non-VTE cohort mean 81.1 years ± 2.23; VTE cohort mean 77.4 years ± 3.45; p = 0.96). Thirty-two events occurred during the surveillance period at our institution; 9 individuals had more than one VTE. At the time of VTE, 80% had hyperparathyroidism (mean PTH ± SD; 97.56 pg/mL ± 90.76), 21% had hyperprolactinemia (prolactin 25.7μg/L ± 43.41), 62.5% had hypergastrinemia (mean gastrin 1100.9 pg/mL ± 3127.8), and 84.6% had non-functional pancreatic neuroendocrine tumors. One patient was identified to have a Factor V Leiden mutation, 3 patients had lupus anti-coagulant. Eleven patients experienced events within a post-surgical period of 3 months.

Conclusions: Hypercoagulability in MEN1 has been previously unidentified. Our cohort data suggests a two-fold increase in the incidence of VTE as compared to the general population, with a high risk occurring within the perioperative period. Further mechanistic investigation and validation from other cohorts are needed to confirm the increased prevalence of VTE in this population.

1Heit, John A., et al. Epidemiology of venous thromboembolism. Nat Rev Cardiol 2015 Aug;12(8): 464-474.
SAT-382 Pyridoxal 5’-Phosphate Cerebrospinal Fluid Abnormalities in Hyopophosphatasia Before and After Enzyme Replacement Therapy
Authors: Wright Jordan J , Hu Jiun-Ruey , Yi Zahra Shajani , Dahir Kathryn McCrystal
Published: May 2020

Introduction: Hypophosphatasia (HPP) is an inborn error of metabolism due to deficiency of tissue non-specific alkaline phosphatase (TNSALP), characterized by a wide range of skeletal and neurological symptoms. Little is known about the mechanism of neurologic involvement, but it may result from inefficient hydrolysis of vitamin B6 (pyridoxal 5’-phosphate, PLP), which is involved in neurotransmitter synthesis and is markedly elevated in HPP. However, it remains unknown what PLP levels are in the cerebrospinal fluid (CSF) of patients with HPP. We report two cases: (1) one with pre- and post-treatment CSF PLP levels and (2) one with pre-treatment CSF PLP and neurotransmitter levels.

Case 1: A 30-year-old woman with extensive fracture history presented with seizure-like activity. Laboratory evaluation while on no B6 supplementation was notable for low ALP (14 U/L, ref: 40-150), high plasma PLP (362 mcg/L, ref: 5-50), and high CSF PLP (16 mcg/L), with a low CSF/plasma PLP ratio (0.04, ref: 0.1-0.7[1]). Genetic testing showed a pathogenic variant of TNSALP (Het c.1133A>T, p.Asp378Val, rs 121918008). After initiation of enzyme replacement therapy (ERT), her plasma and CSF PLP levels normalized to 6.3 and 1.4 mcg/L, respectively, with a ratio of 0.23. She reported improvement in energy level and seizure-like activity after several months of ERT.

Case 2: A 34-year-old man with no history of skeletal pathology presented with diffuse musculoskeletal pain, weakness, and loss of sensory function attributed to generalized peripheral neuropathy. Laboratory evaluation while on no B6 supplementation was notable for low ALP (23 U/L), high plasma PLP (496 mcg/L), and high CSF PLP (17 mcg/L) with a low CSF/plasma ratio (0.03). CSF neurotransmitter levels were also low, including 5-hydroxyindoleacetic acid (40 nmol/L, ref: 67-140) and homovanillic acid (115 nmol/L, ref: 145-324). Genetic testing showed heterozygous expression of a catastrophic frameshift mutation of TNSALP (c.662del, p.Gly221Valfs*56). ERT is being considered at this time.

Conclusions: We report the novel findings that in these patients with HPP:

1. CSF PLP levels are markedly elevated, though to a much lesser extent than plasma levels. Elevated CSF/plasma ratios suggest that transport of PLP and its metabolites across the blood brain barrier may be impaired in HPP.

2. Decreased CSF neurotransmitter levels, as seen in our patient, may contribute to neurologic symptoms of HPP.

3. Enzyme replacement therapy normalized both CSF and plasma PLP in case 1. Further studies are needed to understand how these changes affect patient symptoms and prognosis.

Citation:

1. Albersen, M., et al., Vitamin B-6 vitamers in human plasma and cerebrospinal fluid. Am J Clin Nutr, 2014. 100(2): p. 587-92