bluuebunny/crossref_metadata_2025 huggingface dataset:
OR05-4 Case Series of Transmasculine Adolescents Who Developed Polycythemia on Gender Affirming Hormone Therapy
Authors: Latysheva Natalya
Published: November 2022

Study Objective

This case series was designed to describe cases of polycythemia among transgender male adolescent patients undergoing treatment with testosterone.

Methods

Retrospective data was collected for our Pediatric Gender Clinic between Jan 2014 and June 2021. Variables assessed included demographic and laboratory data, presence of comorbidities, testosterone dose and level at the time of polycythemia, concurrent treatment with other hormonal treatments, and duration of therapy. Polycythemia in children was defined as hematocrit (HCT) +2 SD above mean for age and gender. Male reference range was used for HCT after 3 months of treatment.

Results

Seven of 131 patients were identified with polycythemia (5.3%). The mean age was 17.9 ± 3.7 (SD) years at the time of polycythemia and 6 of 7 patients were overweight or obese. All patients were receiving testosterone by injection. The mean testosterone dose was 51.4 ± 13.3 (SD) mg/m2/2wks or 92.9 mg/2wk ±22.1 (SD). All patients developed polycythemia developed at doses below 75 mg/m2/2wks. The mean testosterone level at the time of polycythemia was 588 ± 315.3. (SD) ng/dL and the mean length of treatment before polycythemia occurred was 18.6 ± 6.2 (SD) months; the shortest duration of treatment was 10 months. Two patients have received concurrent treatment with progestin therapy. Obstructive sleep apnea (OSA) was identified in 1 patient.

Conclusion

Polycythemia occurred in 5.3% of our transgender male patients receiving testosterone therapy. This is a known complication of testosterone therapy that may appear with a relatively low dose and after a relatively short duration of treatment. Patient 1Patient 2Patient 3Patient 4*Patient 5Patient 6Patient 7Age at time of polycythemia, years19.116.616.721.816.917.616.6BMI at time of polycythemia, kg/m225.239.634.130.949.619.426.7Smoking/vaping NNNVapingNNNComorbidities: Asthma, OSANAsthma, mild OSAAsthmaNNNNAdditional hormonal therapyNMedroxyprogesterone acetate injectionEtonogestrel implantNNNNHCT level at baseline, %46.544.143.050.444.245.841.1HCT level at the time of polycythemia, %

55.050.150.053.050.550.650.8Testosterone level at the time of polycythemia, ng/dL3997033455254821253416Length of Tx with testosterone, months10141426182523Dose of testosterone, mg/2wk801008060130100100Dose of testosterone, mg/m2/2wk47.369.741.231.249.664.556.5Table 1. Characteristics of patients with polycythemia.* Patient was taking an injectable dose of testosterone for 6-8 months prior to the first visit to our clinic

Presentation: Saturday, June 11, 2022 12:15 p.m. - 12:30 p.m.
PMON193 Reactive oxygen species in the development of gonadal failure in late-onset transaldolase deficiency
Authors: Zangen David , Florsheim Natan , Lavi Eran , Kerem Liya , Renbaum Paul , Cahan Ranit , Libdeh Abdulsalam Abu , Levi-Lahad Efrat , Mendelsohn Espen Eliyahu
Published: November 2022

Background

Deficiency in Transaldolase, an essential enzyme in regulating NADPH and ribose 5-phosphate production, has been reported in only 39 patients to date. Most patients present already prenatally/neonatally with intra uterine growth retardation, hepato(spleno)megaly and consequent liver failure, anemia, thrombocytopenia, dysmorphic facial features, cardiac and skin abnormalities and hypergonadotropic hypogonadism in cases of late-onset presentation. The mechanism underlying gonadal dysfunction is not fully understood.

Clinical case, methods and results

A 15y old male from an Indian-Jewish background, with atrial-septal-defect, renal tubulopathy, mild facial dysmorphism and transient liver enzymes abnormalities presented with absence of pubertal development. His gonadotropins levels were elevated (LH – 22 IU/L, FSH 103 IU/L) while testosterone was low. Whole exome sequencing revealed homozygous variant in a novel T167M missense mutation in TALDO1 (transaldolase) gene whereas both parents were heterozygote carriers. Proteine structure analysis indicate that Threonine 167 is part of transaldolase catalytic site and critical for its function. As transaldolase activity is required for nucleic acids production, NADPH synthesis and reduction of cellular Reactive Oxygen Species (ROS) we measured the ROS accumulation using 2′,7′-dichlorodihydrofluorescein diacetate, in fibroblasts derived from the patient and normal controls. The affected patient's fibroblasts showed a significantly increased accumulation of ROS (207%, P=0.007), compared to controls indicating an increased susceptibility to oxidative stress.

Conclusion

The novel T167M missense mutation in the transaldolase (TALDO1) gene causes a unique clinical presentation including a relative mild liver involvement and hypergonadotrophic hypogonadism. The compromised TALDO1 activity in preventing cellular ROS accumulation in actual patient's fibroblasts is shown here for the first time and may indicate the therapeutic use of antioxidants in all patients with TALDO1 dysfunction. The high expression of TALDO1 in the testis and the gonadal failure suggests its tissue specific requirement in preventing the accumulation of ROS during steroidogenesis.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON307 Adipokines and Bone Health in Hispanic Children
Authors: Shawar Reem , Ayash Heba E , Puyua Maurice , Bacha Fida
Published: November 2022

Introduction

Leptin and adiponectin are adipocytokines with important metabolic functions. Leptin to adiponectin ratio is a marker of adipose tissue dysfunction and is associated with insulin resistance and cardiometabolic risk. We previously reported a negative effect of adiposity on bone health. We examined the relationship of adipokines to bone mass in a large cohort of Hispanic youth. We hypothesized that adipocytokine dysregulation in childhood may contribute to adverse bone health in children with obesity.

Methods

We studied 501 (237 males and 264 females) children and adolescents from the Viva la Familia Study. They were all pubertal; mean age (SD) 14.1 (2.5) years; 27% were normal weight (NW), 20% overweight (OW) and 53% obese (OB). They underwent measurement of body composition, total body less head bone mineral content (BMC) and density (BMD) by DXA scan; fasting leptin and adiponectin. The leptin to adiponectin ratio was calculated.

Results

Leptin concentration and leptin to adiponectin ratio were higher in OW and OB compared to NW (p<.001) whereas adiponectin was lower in OW and OB compared to NW (p<.001). After adjusting for sex, age, Tanner stage and lean mass, BMC was inversely related to leptin (r = -0.15, p =.001) and leptin to adiponectin ratio (r = -0.20, p <.001) but positively related to adiponectin (r = 0.12, p =.007). Similar relationships were found for BMD.

In a regression model with BMC as the dependent variable, together and independently of sex, age, Tanner stage, lean mass and fat mass, leptin to adiponectin ratio negatively contributed (β = -0.060, p=.002) to the variance of BMC (R2 = 0.877, p <.001). Similar findings were seen with BMD as the dependent variable.

Conclusion

Elevated leptin to adiponectin ratio, a manifestation of adipokine dysregulation, may contribute negatively to bone health (BMC and BMD) in children with overweight/obesity.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON325 Rare Presentation of Precocious Puberty Secondary to LH-Secreting Adenoma
Authors: Uhing Anna , Salamat Shahriar , Chen Melinda
Published: November 2022

Background

Normal male puberty begins after 9 years of age with a concurrent rise in luteinizing hormone (LH) and follicle-stimulating hormone (FSH). These play distinct roles, with LH acting on Leydig cells to stimulate testosterone and secondary sex characteristics, while FSH promotes testicular growth by stimulating the maturation of seminiferous tubules.

Clinical Case

An 8-year 9-month-old male TV presented with a 1.5 year history of facial hair and 9 months of phallic growth, body odor and acne. Physical exam revealed phallic enlargement, Tanner stage I pubic hair and firm, borderline pubertal testes (4 cc bilaterally). Bone age was 9 years at a chronological age of 8 years 6 months. Laboratory evaluation was inconsistent with typical puberty – LH (9 mIU/mL, RR 0.0-0.3 mIU/mL) and testosterone (519 ng/dL, RR 2-8 ng/dL) were elevated while FSH was pre-pubertal (<0.1 mIU/mL, RR 0.0-2.8 mIU/mL) and androgens were only mildly elevated (androstenedione 0.363 ng/mL, RR 0.03-0.3; 17-OHP 155 ng/dL, RR<63 ng/dL). Head MRI revealed an anterior pituitary adenoma measuring 8×12×10 mm. After failing to respond to leuprolide, TV was initiated on spironolactone and anastrozole to minimize pubertal progression prior to transsphenoidal adenomectomy. Surgical pathology was positive for steroidogenic factor 1 but negative for LH, FSH and thyroid-stimulating hormone immunoreactivity. However, he had post-operative reduction of LH (0.4 mIU/mL, RR 0.0-0.3 mIU/mL) and testosterone (16 ng/dL, RR 2-8 ng/dL).

Conclusions

Normal male puberty follows a predictable sequence, beginning with testicular enlargement followed by body odor, pubic hair growth, phallic enlargement, voice change, and increased growth velocity. In TV, hypersecretion of LH led to testosterone production, causing secondary sex characteristics. However, his pre-pubertal FSH did not stimulate the testicular growth that typically heralds the onset of puberty. Digression from typical pubertal sequence and isolated high LH raised concern for a pathologic cause. Central precocious puberty (CPP) is most often idiopathic in females, although more commonly associated with CNS lesions in males. Rarely, precocious puberty results from a functioning gonadotroph adenoma. Less than 1% of these are hormonally active. If active, these more often secrete FSH or co-secrete FSH and LH, and typically require surgical resection given limited success of medical therapies. This case illustrates the distinct roles of FSH and LH in pubertal development. Departures from the typical sequence of development should expand one's differential to include etiologies resulting in nonconcurrent secretion of gonadotropins.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PSAT118 Elevated Serum IgA at Onset of Type 1 Diabetes in Children
Authors: Thakkar Amruta , Redondo Maria , Hajjar Joud , Minard Charles , Huang Xiaofan , Hwu Kathy
Published: November 2022

IgA is the most produced antibody in the body. There are two major forms of IgA, monomeric serum IgA and dimeric mucosal (secretory) IgA. During infection or inflammation, IgA binding to its antigen induces crosslinking of FcaRI, which activates immune effector cells to carry out their functions and can thus induce pro-inflammatory responses, which is hypothesized to occur when IgA titers are high. (1,2) Elevated serum IgA has been associated with autoimmune diseases (3) including type 1 diabetes (T1D) but its role in disease pathogenesis is not well known. To address this gap in knowledge, we aimed to test the hypothesis that at diagnosis of T1D, children and adolescents with elevated serum IgA have different demographic, clinical and laboratory characteristics than those with normal serum IgA.

We analyzed 612 children 9.7 [4.2] (mean [SD]) years old (59% Non-Hispanic White, 20% Hispanic, 16% Black, 5% Other) with new onset T1D diagnosed between 1/2008-2/2012 at a large, academic pediatric hospital. Serum IgA values above age-adjusted normal ranges were defined as elevated serum IgA. Patients with low IgA were excluded because of known association between IgA deficiency and autoimmune disease. Demographic and clinical variables were compared between youth with and without elevated serum IgA. Univariable and multivariable regression models were used to compare baseline characteristics that were associated with elevated serum IgA. A significance level of 0.05 was used.

We found that elevated serum IgA was present in 21% (128/612) of the children at T1D onset. By multivariate analysis, compared to children with normal IgA, those with elevated IgA were more likely to be of Hispanic ethnicity (Odds Ratio (OR)=3.27, 95% Confidence Interval (CI)=2.00, 5.35) and had a higher hemoglobin A1c (A1c) (OR=1.13, 95% CI=1.01, 1.26). They also had higher odds of having insulin autoantibody (IAA) positivity at diagnosis (OR=1.65, 95% CI=1.10, 2.45) and lower odds of GAD autoantibody (GADA) positivity (OR= 0.57, 95% CI=0.36, 0.93) (all, p<0.05), while there were no significant associations with age, glucose level or presence of DKA.

In sum, at diagnosis of T1D, 21% of youth have elevated serum IgA and this was associated with Hispanic ethnicity, higher A1C and positivity for IAA. Our findings suggest that elevated serum IgA may help identify youth with T1D, islet autoimmunity and insulin deficiency. Further studies are warranted to investigate potential pathogenic correlates and persistence over time. This knowledge may lead to new targeted strategies to preserve or improve beta-cell function in individuals with clinical or pre-clinical T1D.

References

(1) Hansen IS et al. Cell Mol Life Sci. 2019 Mar;76(6): 1041-1055. PMID: 30498997 (2) Leong KW et al. DNA Cell Biol. 2014 Dec;33(12): 823-9. PMID: 25188736 (3) Kerr MA. Biochem J. 1990 Oct 15;271(2): 285-96. PMID: 2241915

Presentation: Saturday, June 11, 2022 1:00 p.m. - 3:00 p.m.
LBMON272 Chromatoid Bodies In The Regulation Of Spermatogenesis: Novel Role Of Grth
Authors: Anbazhagan Rajakumar , Kavarthapu Raghuveer , Dufau Maria L
Published: November 2022

More than 5 million couples in US are affected by infertility. Male infertility is a major cause (>30%), and understanding spermatogenesis is important to address the problem. During early spermatogenesis, mRNAs are actively transcribed, transported and stored transiently in chromatoid bodies (CBs). CBs are membrane-less perinuclear organelles which serve as storehouses of mRNAs until later stages of spermatogenesis. Gonadotropin-regulated testicular RNA helicase (GRTH/DDX25) plays critical role in spermatogenesis. In addition to its inherent helicase activity, GRTH transports mRNAs from nucleus to cytoplasm and from there to CBs for storage until translation. In Germ cells, there are two species of GRTH, the 56 kDa non-phospho and 61 kDa phospho (pGRTH) forms. Our early studies revealed a missense mutation (R242H) of GRTH in Japanese azoospermic men which resulted in the lack of pGRTH in in vitro studies. GRTH knock-in (KI) mice with the human mutant GRTH gene (R242H), show loss of the phospho-species from cytoplasm and CBs with preservation of the non-phospho form in the cytoplasm, Nucleus and CBs. GRTH-KI mice are sterile, lack elongated spermatids and spermatozoa with spermatogenic arrest at step 8 of round spermatids (RS). CBs isolated from GRTH KI mice are smaller, highly condensed, lack pGRTH and RNA-Seq analysis of isolated mRNA (from CBs) revealed differential abundance of mRNAs related to spermatid differentiation. Integrative analysis of small RNA and mRNA expression will provide cohesive outlook of molecular mechanisms of gene regulation during spermatogenesis. Hence, we proceeded with small RNAseq analysis which reveal differential expression (Padj<0. 05) of piRNAs and miRNAs. In total 226706 piRNAs upregulated and 166567 piRNAs downregulated in the round spermatids. Several miRNAs like miR32, miR7, miR184, miR335, miR140, miR141, miR1981, miR202, miR880, miR669c were downregulated. These are involved in the positive regulation of spermatid differentiation, sperm motility, mRNA processing and decay. The miRNAs, miR150, miR196a-2, miR7b, miR652, miR146, miR10b, miR379, miR122a, miR27a, miR127 and miR328 were upregulated, which negatively regulate the spermatid differentiation. Gene ontology analysis revealed that the majority of downregulated miRNAs belong to regulation of gene expression. Majority of upregulated miRNAs belong to miRNA mediated suppression of translation mostly associated with spermiogenesis and CBs. The identified miRNAs regulate essential genes like Tnp2, TSSK6, Pabpc1, Ppp1cc and UPF2 etc. These indicates that the CBs mediate highly specialized mRNA metabolism process and serve as a processing center of essential mRNAs. These results demonstrate the importance of GRTH and phospho-GRTH in the regulation of CB and their role in mRNA storage and stability of germ cell specific mRNAs during spermiogenesis and healthy sperm formation.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
LBODP090 The Addition Of Anastrozole To Standard Testosterone Enanthate Treatment Significantly Improves Penile Size In Adolescent Boys With Micropenis
Authors: Papadimitriou Dimitrios T , Dermitzaki Eleni , Papagianni Maria , Papadimitriou Anastasios , Vlahos Nikos F , Mastorakos George
Published: November 2022

Background

Micropenis is treated preferably in infancy (≤ 2 yrs) or at the onset οf puberty, usually with 3 (2-4) monthly testosterone enanthate I. M. injections at the dose of 100 mg/m2. This short-term therapy may temporarily advance bone maturation but with a concomitant increase in height velocity and no apparent change in predicted adult height. Bone maturation depends on locally produced estrogens by aromatization. Third generation aromatase inhibitors (AIs) are used as an off-label treatment to improve predicted adult height (PAH) in boys as well as in girls, either as monotherapy or in combination with growth hormone and/or puberty inhibition. They induce reverse binding inhibiting the activity of aromatase (a cytochrome P450 enzyme), which catalyzes the conversion of androstenedione and testosterone to estrone and estradiol, respectively, resulting in a substantial increase of the circulating testosterone concentrations.

Aims

To compare the traditional treatment of isolated - idiopathic - relative micropenis in boys with testosterone enanthate monotherapy to its combination with anastrozole 1 mg×1 p. o.

Methods

164 boys with micropenis (stretched penile length ≤ -2 SD) received testosterone enanthate 100 mg/m2 I. M. /month either as monotherapy (n=63, mean age 10.8 yrs, group A) or in combination with anastrozole 1 mg/day (n=101, mean age 11 yrs, group B) for 3 months. Stretched penile length, bone maturation and auxological data were analyzed. All measurements were performed by the same examiner. The choice of therapeutic intervention was made randomly. Groups A and B did not differ in terms of age at intervention onset, bone age, target height or predicted adult height. They underwent a 6-month follow-up that included clinical examination, bone age X-ray evaluated by BoneXpert ver. 3.2. 0 (Visiana, Denmark), and laboratory tests at 8: 00hrs (LH, FSH, testosterone, estradiol, estrone), prior and under treatment.

Results

In both groups penile length normalized: for group A gain was +1.9 cm (+2. 08 SD) and for group B +2.24 cm (+2.3 SD), with group B attaining a greater length by +18% (p=0. 004) due to the higher testosterone concentrations attained by at least 50%. Group A presented a slight acceleration of height velocity with parallel advancement of their bone age maturation while group B with unchanged or lower estradiol and estrone concentrations maintained their height velocity with parallel movement of their bone age maturation.

Conclusions

Addition of anastrozole 1 mg/day p. o. in testosterone enanthate treatment for idiopathic-isolated-relative micropenis at the beginning of puberty significantly improves penile length by almost 20% while the tempo of height velocity and bone maturation continue their previous track.

Presentation: No date and time listed
ODP401 A new FGFR1 mutation causing Idiopathic Hypogonadotropic Hypogonadism and Polydactyly
Authors: Espinosa Luis Borges , Ayala Alejandro , Dicenso Daniela
Published: November 2022

Introduction

Congenital idiopathic hypogonadotropic hypogonadism (IHH) is a heterogeneous condition with unknown genetic basis in approximately 30% of patients. We describe a case with normosmic IHH and an unreported FGFR1 mutation.

Case Report

A 23-year-old hispanic man, with a history of polydactyly presented to our clinic for evaluation of delayed puberty. He had no facial hair and minimal pubic and axillary hair. He reported fatigue, low libido and scant ejaculation. He denied anosmia, eating disorders, learning disability, urological malformations, obesity, galactorrhea, testicular trauma, mumps, or use of androgenic steroids. Family history was significant for polydactyly in his mother. Physical exam findings showed normal vitals, height 185 cm, BMI 24 kg/m2, eunuchoid body habitus, bilateral gynecomastia, no facial hair, scant axillary and pubic hair, prepubertal 2 ml testicles within the scrotum and a micro-penis. Serum total testosterone was 7 ng/dl (250-1100 ng/dl), free testosterone 0.9 pg/ml (9.3-26.5 pg/ml), FSH 2.2 mIU/ml (1.6-8. 0 mIU/ml), LH 0.8 mIU/ml (1.5-9.3 mIU/ml), Prolactin 4.1 ng/ml (4-15.2 ng/ml), TSH 1.68 mIU/ml (0.4-5.5 mIU/ml), free t4 1.2 ng/dl (0.8-1.7 ng/dl), growth hormone 0.3 ng/ml (0. 0-10 ng/ml), IGF-1 175 ng/ml (155-432 ng/ml), ACTH 14.6 pg/ml (7.2-63 pg/ml), cortisol (not am) 3. 0 ug/dl (6.2-19.4 ug/dl). Pituitary MRI revealed no focal sellar/suprasellar lesions with normal enhancement of the pituitary gland . DXA scan showed a low bone density for his age with Z-scores below -2. 0 in all regions. Chromosome analysis revealed a normal male 46, XY karyotype. Genetic testing of both patient and mother, using Invitae test kit, revealed an heterozygous mutation in FGFR1 gene, exon 9, variant c.1231C>A (p. Gln411Lys), not described in medical literature. A diagnosis of normosmic idiopathic hypogonadotropic hypogonadism (nIHH) was made. Testosterone cypionate IM 100 mg once weekly was initiated and after 6 months he experienced improvement in energy, libido, muscle mass, facial and body hair, and virilization of his voice.

Conclusions

The heterogeneity in phenotypic presentation of IHH makes its diagnosis a challenge. Furthermore, the genetic basis for these multiple phenotypes are still not fully described. We report a new FGFR1 mutation causing hypogonadism. Reference: Ferreira Gontijo, L., Latronico, A. Approach to the Patient With Hypogonadotropic Hypogonadism. JES, Vol. 98, Issue 5, 1 May 2013, P. 1781–1788.

Presentation: No date and time listed
ODP412 High Frequency of Erythrocytosis in Hypogonadal Men on Testosterone Replacement Therapy
Authors: Pérez-Luis Jesús , Adolewska Patrycja Dagmara
Published: November 2022

Introduction

Erythrocytosis is a frequent side effect of testosterone replacement therapy (TRT) in hypogonadal males and can be deleterious by increasing blood viscosity, which can lead to headaches, hypertension, and thromboembolic complications. Main causes of erythrocytosis are short-acting testosterone injections, smoking habit and obesity.

Aim

Study the frequency of erythrocytosis induced by TRT and its predictive factors in hypogonadal men in our setting. Subjects and methods: A retrospective study in hypogonadal males on TRT for at least 3 months was conducted. Age, age at treatment onset, months on treatment, initial hematocrit (IHt), highest hematocrit during follow-up (MaxHt), hematocrit on last visit (FHt), hematocrit increase (HtIn), maximum hematocrit increase (HtMaxIn), anthropometric measures, sex hormones and biochemical parameters on the last visit were recorded. In addition, testosterone formulation, smoking status, dyslipidemia, hypertension, diabetes, type of hypogonadism and presence of cardiovascular disease (CVD) were also documented. A hematocrit ≥ 50% (Ht≥50) was considered erythrocytosis and one ≥ 53% was indication for phlebotomy (Ht≥53).

Results

108 patients were included, age 54 ± 19 years, age at THT onset 44 ± 18 years, BMI 30 ± 5 Kg/m 2 . On the last visit, 29% had FHt≥50 and 11% had FHt≥53, while during follow-up 58% had MaxHt≥50 and 35% had MaxHt≥53. Moreover, 18% were smokers, 61% had dyslipidemia, 48% had hypertension, 36% were diabetic, 59% were on short acting IM testosterone injections (cypionate), 25% were on extended-release IM testosterone injections (undecanoate) and 16% were on transdermal testosterone. On the other hand, 13% had CVD at onset and 6% developed CVD during follow-up. None of the hematocrit or hematocrit increase values was related to the type of testosterone formulation, smoking habit or appearance of CVD during follow-up. They also did not correlate with serum total testosterone, other sexual hormones and lipids levels or BMI. Interestingly, there was no difference in total testosterone levels between the three testosterone preparations groups. FHt (44 vs 48%, p=0,018) and HtIn values (1.86 vs 5.31%, p=0,036) were lower in those with previous CVD disease. This fact cannot be explained by BMI since there was no difference between patients with and without CVD. Although it could be due to a lower percentage of smokers in the group of patients with CVD, that could not be demonstrated.

Discussion

The occurrence of erythrocytosis secondary to TRT in hypogonadal men in our setting was very common and higher than reported previously: 58% of patients during follow-up and 35% required phlebotomy. Hematocrit and hematocrit increase values were independent of BMI, smoking status, type of testosterone formulation and presence of CVD. This lack of association could be due in part to the small number of individuals in some of the subgroups.

Presentation: No date and time listed
ODP058 Topiramate causing elevation of urinary cortisol assay
Authors: Grissett Bryan , Sosa-Pagan Max , Iqbal Qasim Zafar
Published: November 2022

69-year-old female with pituitary adenoma status-post transsphenoidal resection 6 years prior and isolated growth hormone deficiency [on somatropin 0.2 mg daily], migraines [on topiramate 25 mg daily], left lipid-rich adrenal incidentaloma, primary hypothyroidism [on levothyroxine 75 mcg daily], and secondary hyperparathyroidism from vitamin-D deficiency [on ergocalciferol 50000 IU weekly] and hydrochlorothiazide 25 mg daily presented for follow-up. Physical exam: afebrile, blood pressure hypertensive at 163/71 mmHg, heart rate 91 beats per minute, respiratory rate 18 breaths per minute, saturating 100% on room air and patient weighed 105 kg. Patient appeared obese but did not exhibit cervicodorsal fat pads, abdominal striae, or other cushingoid features. Neck was supple and did not demonstrate palpable nodules or lymphadenopathy. Labs ordered at clinic visit demonstrated normal TSH 1.28 with normal free T4 1.21, elevated PTH 232 with corrected calcium 9.7 (on hydrochlorothiazide) and normal vitamin-D, 25 level 43, normal ACTH 21, and normal IGF-1 level 75. Given adrenal incidentaloma and prior pituitary adenoma, the integrity of the hypothalamus-pituitary-adrenal axis was also tested. A subsequent 8: 00 a. m. cortisol was insufficiently suppressed at 5.2 after 1 mg dexamethasone suppression test which raised concern for hypercortisolism from pituitary adenoma or adrenal adenoma. Accordingly, ACTH as well as DHEA-S and 24 hour urine cortisol as well as midnight salivary cortisol level was ordered to assess pituitary versus adrenal origin, respectively. Interestingly, while midnight salivary cortisol was found to be normal at less than 50, the 24 hour urine cortisol measurement was elevated at 878 despite otherwise normal cortisol findings. ACTH and DHEA-S levels are still pending. Plan was made to check two additional midnight cortisol levels and await ACTH and DHEA-S levels. In an effort to explain the urinary cortisol elevation, the patient's medications were reviewed and subsequent literature search revealed that topiramate may interfere with urinary cortisol measurement by causing a dose-dependent increase in the measured urinary cortisol, presumably due to its action on the kidney during renal excretion. Patient had been on topiramate for approximately 6 months prior to this assay.

Presentation: No date and time listed
ODP415 Inclusion Health Clinic: Gender-Affirming Hormone Therapy & Surgery Booklet and Review of Efficacy in Patient-Provider Encounters
Authors: Pulsiano Sune Wolf , Sarvaideo Jenna
Published: November 2022

While gender-inclusive health has been a growing field, comprehensive information on the topic has lacked for both patients and providers. The "Inclusion Health Clinic (IHC) Gender-Affirming Hormone Therapy & Surgery" booklet, modelled after other Froedtert Health resources and created using guidelines from Endocrine Society, University of California, San Francisco, and World Professional Association for Transgender Health (WPATH), was introduced to IHC on April 28, 2021. Its goal was to facilitate patient-provider relationships through sharing clinical knowledge and practice preferences with patients as well as to assist providers in communicating concepts to individuals seeking advice and care. Booklet efficacy was analyzed through anonymous surveys self-selected among transgender and gender-nonconforming patients seen at Inclusion Health Clinic. Regardless of gender identity or new patient status, 100% of survey respondents found the booklet useful in generating a productive conversation. For numerical questions, averages for all data sets were above 4. 0/5. 0: ease of navigation was rated highest (4.75 +/- .24), followed by scope of care (4.25 +/- .34), and lastly the booklet's utility in explaining concepts (4.15 +/- .36). For qualitative responses, respondents found the most value in pages related to hormone therapy and surgery. Data from the survey was primarily used to improve the booklet; this revision is projected to be distributed to IHC in Spring 2022. The aspiration is for the booklet to evolve as care for the LGBTQ+ population advances, and that it will continue to serve as a relevant and useful source of information on gender-affirming health needs.

Presentation: No date and time listed
ODP419 Measurement of Free Testosterone in Serum using Equilibrium Dialysis Coupled with ID-UHPLC-MS/MS: in Comparison to Calculated FT
Authors: Zhou Hui , Ribera Ashley , Danilenko Uliana , Sugahara Otoe , Vesper Hubert
Published: November 2022

Assessment of free testosterone (FT) has been recommended in recent clinical practice guidelines as a biomarker for diagnosis and management of testosterone related disorders such as male testosterone deficiency due to hypogonadism and female androgen excess due to polycystic ovary syndrome (PCOS). FT is currently measured using direct commercial immunoassays, assays using equilibrium dialysis (ED) or other separation techniques, or estimated by calculating free testosterone (cFT) using different formulas. The most widely used formula was developed by Vermeulen et al and uses total testosterone and sex hormone binding globulin (SHBG) concentrations. The ED approach is recognized as the gold standard; however, it is technically challenging for routine clinical use. CDC's Clinical Standardization Program (CDC CSP) has developed a high throughput method using ED coupled with isotope dilution ultra-high-performance liquid chromatography tandem mass spectrometry. The study to evaluate the agreement of this ED-based method to the cFT using different algorithms is being conducted. Serum samples are dialyzed in a custom-designed multi-well plate against a protein-free HEPES buffer (pH 7.4) at 37 °C until equilibrium. After isolating endogenous FT from protein-bound testosterone by ED, isotope-labeled internal standard (13C3-testosterone) was added to the resulting dialysate for quantification. Certified pure primary reference material (National Measurement Institute-M914) was used to prepare calibrators, enabling traceable quantitation and ensuring measurement trueness. FT was further isolated from the dialysate matrix using supported liquid extraction and a chromatographic separation from interfering compounds and quantitation by tandem MS. The assay allowed detection of 2.9 pg/mL FT, with the bias within ± 5% and precision less than 10% CV. A total of 45 samples with a wide range of TT (21-912 ng/dL) and SHBG (11-129 nmol/L) were analyzed. The obtained FT values were compared to the cFT values obtained with the Vermeulen algorithm. The mean difference of 34.1% (95% CI 29.1% to 39.9%) between the measured and calculated FT were observed, with majority of the calculated values overestimating FT concentrations. The described high throughput method for FT allows for sufficiently accurate and precise measurement and can provide more reliable FT measurements compared to cFT (that can lead to overestimation of FT) for routine applications including large epidemiologic studies.

Presentation: No date and time listed
PMON226 Primary Ovarian Insufficiency: A Case of a Missed Puberty
Authors: Lin Jack , Suarez Maria Gabriella
Published: November 2022

Premature Ovarian Insufficiency (POI) is a loss of ovarian function in women less than 40 years old. We present a case of a female with primary amenorrhea due to autoimmune primary ovarian insufficiency (APOI), in which diagnosis was delayed as she had normal breast and pubic/axillary hair development.

A 18-year-old female presented to endocrine clinic for evaluation of primary amenorrhea and hypoplastic uterus and ovaries. During her regular follow ups with her pediatrician it was noted that she had not undergone menarche but since her breast development was normal, further evaluation was not pursued. She had a complicated social history and missed several of her follow up appointments. However, further evaluation was done after she presented with severe pain during intercourse. During her physical exam, it was found she had a vaginal opening of 4 cm with an unpalpable cervix. She did however have pubic/axillary hair and breast development consistent with Tanner stage V. Pelvic MRI showed hypoplastic uterus/ovaries. Her lab work revealed she had a total testosterone of 28ng/dL, prolactin 11.1ng/mL, TSH 1.68iIU/mL, LH 42.2mIU/mL, FSH99.77 mIU/mL, estradiol29.36 pg/ML, AMH 3.1ng/mL. Her karyotype was 46XX. Testing for Fragile×Syndrome and 21 hydroxylase antibodies were negative. Anti-ovarian antibody (AOA) was obtained and found to be positive (1: 160). She was diagnosed with APOI and started on estrogen with a goal to increase to adult doses over 1.5-2 years. Progestins are planned to be introduced once she reaches adult doses of estrogen or develops breakthrough bleeding.

Typical presentation of patients with POI is primary amenorrhea and lack of breast development during puberty. However, it is important to remember that there can be a broad spectrum of presenting symptoms with variable breast, uterine, and ovarian development as not all patients have the same degree of estrogen deficiency. This is a case of a patient with POI with hypoplastic uterus and ovaries who had Tanner V breast development. Given this patients ovarian atrophy it is likely that the majority of estrogen production originated from adrenal glands and fat cells, which contributed to the normal development of secondary sexual characteristics delaying her diagnosis. Therefore, identification of abnormal menstrual patterns in adolescence should always raise concern for POI even in the setting of normal breast and pubic/axillary hair development.

POI is due to premature deprivation of estrogen with subsequent amenorrhea and compensatory elevations in FSH/LH. Early diagnosis of this condition can prevent potential health complications in such as decreased bone mineral density, psychological complications, and extremely painful intercourse as our patient had. It is important to note that fertility in patients with POI is also unpredictable, however earlier diagnosis/treatment can improve fertility preservation, especially in women with normal AMH levels such as this patient.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON243 Metabolic Disorders Genetically Modify Polycystic Ovary Syndrome through its Candidate Hormones
Authors: Actkins Ky’Era , Davis Lea
Published: November 2022

Introduction

As the most common endocrine disorder in reproductive-aged women, polycystic ovary syndrome (PCOS) increases the risk of developing several cardiometabolic disorders like insulin resistance. However, Mendelian randomization (MR) studies provide little support for direct causal associations between PCOS and metabolic conditions with the exception of obesity and type 2 diabetes (T2D). As a result, secondary pathways and biological mediators increasingly emerge as a potential explanation for underlying mechanisms between traits.

Objective

To identify biomarkers that modify PCOS presentation and its comorbid conditions through MR, which uses significant genetic variants from genome-wide association studies to estimate causal effects between variables.

Methods

A bidirectional MR analysis was first implemented to identify associations between PCOS and metabolic traits (i.e., body mass index (BMI), T2D, blood pressure, coronary artery disease, and heart failure), with PCOS acting first as the exposure, then as the outcome. Next, MR was applied between significant metabolic phenotypes from the first analysis and thirty-two biomarkers from UK Biobank to identify any overlap with biomarkers that also significantly increased risk for PCOS. Mirroring biomarkers from a hospital population were used as a replication dataset and were further analyzed in a mediation analysis with clinical laboratory measurements.

Results

Testosterone from females (β = 0.83, p = 4.67e-04) and sex hormone-binding globulin (SHBG, β = -0.01, p = 5.00e-06) were identified as significant exposures for PCOS. These biomarkers also exhibited causal effects on BMI (βFT = 0.10, p = 5.81e-12; βSHBG = 0.001, p = 4.56e-10) and T2D (βSHBG = -0.004, p = 4.99e-26), which were the only two phenotypes that exerted causal risk for PCOS (p < 0.001). Although SHBG had strong bidirectional effects on both traits, female-derived testosterone was the only testosterone exposure (vs sex-combined and male testosterone) that increased BMI (β = 0.04, p = 1.08e-16). This positive association with BMI was exclusively replicated for free testosterone as the outcome. Although no significant associations were observed in the mediation analysis for BMI and free testosterone, total testosterone mediated 29% of the genetic pathway from T2D to PCOS.

Conclusions

Overall, MR can expand our understanding on the phenotypic irregularities of PCOS through examination of biomarker etiology cross-traits. These results imply a strong interaction between prevalent PCOS characteristics, one that may mitigate the advancement of other chronic comorbidities. Therefore, further research is needed to elucidate PCOS regulation, which may reveal a more complex web of modifiable effects that have yet to be explored.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON251 Loss of Constitutive Androstane Receptor, CAR reduces the serum androgens levels and alters reproductive function
Authors: Asokakumar Anjana , Hernandez Juan P , Xiao Rui , Moore David D , Anakk Sayeepriyadarshini
Published: November 2022

The nuclear receptor, Constitutive Androstane Receptor (CAR/NR1i3) is extensively known for its detoxification function in the liver. We have identified the role of CAR in regulating androgen levels and regulating testis function. Although the inactivation of CAR by the sex hormone axis via androstanol has been known for decades, its role in regulating reproduction has not been addressed. On examining the WT mice and the global CARKO mice, we found that CARKO mice have reduced serum testosterone and androstanol levels. Further, the levels of the steroidogenic gene, Srd5a1 was significantly reduced. This result correlated well with reduced anogenital distance in males, increased anxiety and subfertility. However, we did not find differences in the expression of detoxification genes in the CARKO testis. Histological analysis of the testis of the CARKO mice revealed abnormal Leydig cells and Oil Red O staining showed lipid accumulation in the Leydig cell island. We are currently examining the effect of the reduced androgens in the hypothalamic-pituitary-gonadal axis in the CARKO mice model.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
PMON255 Inflammation and Oxidative Stress in Seminal Plasma: Role of Free Light Chains of Immunoglobulins and Antioxidants as Biomarkers in Male Infertility
Authors: Bruno Carmine , Vergani Edoardo , Oliva Alessandro , Basile Umberto , Napodano Cecilia , Gulli Francesca , Silvestrini Andrea , Meucci Elisabetta , Orlando Patrick , Silvestri Sonia , Tiano Luca , Mancini Antonio
Published: November 2022

Oxidative and inflammatory damage are underlying mechanisms in various conditions related to male infertility, including varicocele. Free light chains of immunoglobulins (FLCs) are considered markers of low-grade inflammation in various conditions. Nowadays it is known that they may play enzymatic and proteolytic activities; they may activate complement cascade and mast cells, with crucial role in the development of contact sensibility; they may also inhibit the autonomous signaling ability of the B-cell receptor. Their detection in biological fluids may be useful as biomarker of inflammation or altered immune response. Coenzyme Q10 (CoQ10), a lipidic antioxidant and component of the mitochondrial respiratory chain, is involved in spermatozoa energy metabolism and motility.

Therefore, we aimed to evaluate FLCs seminal levels in patients with varicocele in comparison to control subjects and to correlate them with CoQ10 and Total Antioxidant Capacity (TAC) in human semen.

A total of 65 patients were enrolled, aged 18-50 years. Semen analysis was performed; patients were divided into three groups: controls (CTRL), 12 normozoospermic patients aged 34 (33-41) years; varicocele (VAR), 29 patients aged 33 (26-37) years; idiopathic (IDIO), 24 oligo-, astheno- and oligoasthenozoospermic patients aged 37 (33.5-40.5) years. FLCs were assayed by turbidimetric method; CoQ10 by HPLC; TAC by spectrophotometric method.

Λ FLCs showed a trend towards higher levels in VAR vs CTRL and IDIO. VAR showed a trend towards lower κ FLCs levels vs the other two groups. Comparing κ/λ ratio VAR showed significantly lower levels vs CTRL and IDIO (Mean±SEM 3.18±0.75; 8.28±1.71; 8.90±2.94, respectively; p<0.05). CoQ10 seminal levels showed higher levels in VAR and IDIO compared to CTRL (57.7±4.13; 65.50±6.95 and 38.30±3.35 mcg/ml, respectively; p<0.05) No different TAC values were found between groups.

Our data confirm lower levels of κ/λ ratio in VAR as previously reported and suggest a possible application as clinical biomarkers for male infertility.

Presentation: Monday, June 13, 2022 12:30 p.m. - 2:30 p.m.
OR07-3 Hormone-Controlled Cooperative Binding of Transcription Factors Within Enhancer Clusters Drives Synergistic Induction of Fasting-Regulated Genes
Authors: Charni-Natan Meital , Goldberg Dana , Goldstein Ido
Published: November 2022

During fasting, hepatocytes produce glucose in response to hormonal signals. Glucagon and glucocorticoids are principal hormones secreted during fasting that cooperate in regulating glucose production via increasing hepatic gluconeogenesis. However, how these hormone signals are integrated in hepatocytes and translated to a biological output is unknown. We used genome-wide profiling of gene expression, enhancer dynamics and transcription factor binding in primary mouse hepatocytes to uncover the mode of cooperation between glucagon and corticosterone (the major glucocorticoid in mice). We found that compared to a single treatment with either hormone alone, a dual treatment initiates a pro-gluconeogenic gene program in hepatocytes by synergistically inducing gluconeogenic genes. We show that the mechanism driving synergistic gene expression is based on glucagon-mediated enhancer activation, leading to increased binding of the glucocorticoid receptor (GR) upon corticosterone stimulation. This was shown by glucagon-dependent increases in the active enhancer mark H3K27ac at GR binding sites (both signals were measured genome-wide via chromatin immunoprecipitation sequencing). Thus, glucagon-dependent enhancer activation, mediated by the glucagon-activated transcription factor CREB, assists GR loading to specific enhancers located near synergistic gluconeogenic genes. This 'assisted loading' mechanism is enhancer-specific as GR was able to efficiently bind other enhancers without assistance by CREB. Interestingly, we found that the glucagon-CREB axis does not only activate single enhancers but is also able to activate entire enhancer clusters. Indeed, CREB binding in one enhancer unit within a cluster, led to cluster-wide enhancer activation, thereby assisting the loading of GR in other enhancer units within the cluster that are unbound by CREB directly. These chromatin and gene expression changes collectively lead to synergistic glucose production from hepatocytes in the presence of glucagon and corticosterone. In summary, we show that hepatocytes integrate extracellular signals by an enhancer-specific mechanism: glucagon activates enhancers, thereby assisting the loading of GR upon stimulation by corticosterone. This chromatin-bound integration of endocrine signals leads to synergistic gene induction and a tailored response to fasting.

Presentation: Saturday, June 11, 2022 12:00 p.m. - 12:15 p.m.
OR26-5 Musashi-2 Regulatory Interactions Promote Adrenal Steroidogenesis
Authors: Baldwin Amber , Mukherjee Neelanjan , Murphy Elisabeth , Sajek Marcin , Walters Kathryn
Published: November 2022

The adrenal gland is the site of steroid hormone synthesis. These hormones regulate many physiological processes including metabolism, blood pressure and volume, and sexual characteristic development. While signaling pathways, steroidogenic enzymes, and transcription factors have all been well characterized, little is known about the contribution of RNA-binding proteins (RBPs) in regulating steroidogenesis. We recently demonstrated that regulated RNA decay controls the timing of the steroidogenic gene expression response and identified 9 RBPs that regulate aldosterone production in an immortalized human cell line model for steroidogenesis, H295R. One of the RBPs we identified was Musashi-2 (MSI2), which is required for development of other steroidogenic tissues such as ovary and testis in mice. We found that MSI2 mRNA is enriched in human adrenal glands relative to other tissues, positively correlates in expression with mRNAs encoding steroidogenic enzymes, and abundantly expressed in H295R cells. Staining of normal human adrenal gland sections demonstrated that MSI2 protein is expressed in all three layers of the human adrenal cortex. Taken together, we hypothesized that MSI2 promotes steroidogenesis through regulating expression of target mRNAs encoding. Using the selective MSI2 inhibitor Ro-08-2750 (Ro) we found that aldosterone and cortisol production is decreased in a dosage dependent manner in H295R cells, which is consistent with siRNA knockdown of MSI2. We performed RNA-immunoprecipitation and sequencing and identified 1428 mRNA targets of MSI2 in H295R cells. A target of interest was Stearoyl-CoA Desaturase (SCD), which functions to convert saturated fatty acids to unsaturated fatty acids that promote formation of lipid droplets. Cholesterol esters in lipid droplets are a source of cholesterol used to produce all steroid hormones. Ro treatment of H295R cells resulted in a suppression of SCD mRNA and protein induction. Furthermore, inhibition of SCD with A939572 caused a dosage dependent decrease in aldosterone production. These data suggest that potentiation of stimulation dependent SCD induction is one mechanism by which MSI2 promotes steroidogenesis. It will be important to determine if and how MSI2 promotes steroidogenesis in primary human adrenal cells and mouse models. Since it is possible to deliver oligonucleotides to the adrenal cortex, disruption of MSI2-SCD mRNA regulatory interactions may open new therapeutic avenues for modulate aldosterone production.

Presentation: Monday, June 13, 2022 12:00 p.m. - 12:15 p.m.