bluuebunny/crossref_metadata_2025 huggingface dataset:
OR08-06 Adjuvant Radiation Therapy Is Not Associated With A Survival Benefit After R0 Resection In Non-Metastatic Adrenocortical Carcinoma
Authors: Lee-Saxton Yeon Joo , Egan Caitlin E , Marshall Teagan E , Tumati Abhinay , Beninato Toni , Zarnegar Rasa , Fahey Thomas J , Finnerty Brendan M
Published: October 2023

Disclosure: Y.J. Lee-Saxton: None. C.E. Egan: None. T.E. Marshall: None. A. Tumati: None. T. Beninato: None. R. Zarnegar: None. T.J. Fahey: None. B.M. Finnerty: None.

Introduction: The benefit of adjuvant radiation therapy (RT) in adrenocortical carcinoma (ACC) is not well characterized, particularly for those who undergo initial R0 surgical resection without evidence of metastatic disease. We aimed to identify factors associated with administration of adjuvant RT in these patients, and to determine the association between adjuvant RT and overall survival in patients with non-metastatic ACC who underwent R0 surgical resection. Methods: The National Cancer Database was queried for histologic codes denoting ACC from 2004-2018. Patients who underwent R0 resection without evidence of metastatic disease were included and placed into two cohorts - those who underwent adjuvant RT and those who did not. Patients with incomplete or unknown data regarding clinically relevant variables were excluded from analysis. A multivariable logistic regression was performed to identify predictors of adjuvant RT administration. A multivariable Cox regression was performed to identify factors associated with overall survival (OS). Kaplan-Meier survival estimates and log-rank test were also performed to assess the impact of adjuvant RT on OS. Results: 388 patients were identified: 51 (13.1%) patients received adjuvant RT, while 337 (86.9%) did not. Adjuvant chemotherapy administration was the only factor associated with administration of adjuvant RT (OR 6.45, 95% CI 3.02 - 13.8, p<0.001). No significant difference was observed between the Kaplan-Meier survival estimates of the two cohorts (p=0.54). After adjusting for patient characteristics including age, sex, co-morbidity index, race, receipt of chemotherapy, and tumor characteristics including tumor size, tumor grade, pathologic tumor stage, and pathologic nodal stage, adjuvant RT was not associated with improved OS in patients who underwent R0 resection (HR 1.01, 95% CI 0.63 - 1.60, p=0.98). However, tumor size >6cm (HR 1.54, 95% CI 1.03 - 2.32, p=0.04), high tumor grade (HR 3.46, 95% CI 1.83 - 6.55, p<0.001), and pathologic N1 stage (HR 2.30, 95% CI 1.06 - 4.94, p=0.03) were associated with worse OS in patients who underwent R0 resection. Conclusions: Treatment with adjuvant RT in patients with ACC who underwent R0 resection was not associated with an OS benefit. Further studies assessing optimal treatment for large and high-grade tumors with lymph node metastases are warranted.

Presentation: Friday, June 16, 2023
FRI410 ESR1 Variants Reduce Estrogen Receptor Alpha Expression In Women With Unexplained Infertility
Authors: Roman Robert A , Liu Haitao , Chorich Lynn Parson , Li Yin , Hall Janet Elizabeth , Diamond Michael P , Korach Kenneth Steven , Layman Lawrence Clarke
Published: October 2023

Disclosure: R.A. Roman: None. H. Liu: None. L.P. Chorich: None. Y. Li: None. J.E. Hall: None. M.P. Diamond: None. K.S. Korach: None. L.C. Layman: None.

Objective: Estrogen receptor alpha (ERα), encoded by ESR1, is vital to human reproduction. DNA sequencing previously identified four ESR1 missense variants of uncertain significance in 197 cisgender women with well-characterized unexplained infertility, including a p.Thr313Met variant with impaired estrogen signaling in vitro. The p.His6Tyr, p.Ser118Pro, and p.Arg269Cys ESR1 variants did not affect estrogen signaling. We hypothesize that milder ESR1 variants could decrease ERα expression in women with unexplained infertility.

Methods: Clinically identified ESR1 variant plasmids were constructed using site-directed mutagenesis and confirmed by Sanger sequencing. HepG2 cells were transiently transfected with either empty vector, wild type (WT) ESR1, p.His6Tyr, p.Ser118Pro, p.Arg269Cys, p.Thr313Met, or p.Gln375His (ESR1 missense variant control) estrogen receptor variant expression plasmids using Lipofectamine 3000 transfection reagent. Cell lysis was isolated from HepG2 cells 24 hours after transfection using RIPA lysis buffer. ERα expression was assessed by western blot fluorescence imaging and normalized to β-actin on LI-COR Odyssey DLx. Quantitative analysis was performed using Empiria Studio Software v.2.

Results: Western blot analysis showed that the ESR1 p.His6Tyr variant had 87% decreased and the p.Ser118Pro variant had 75.6% decreased expression compared to WT ERα in vitro. The p.Arg269Cys, p.Thr313Met, and p.Gln375His variants had comparable expression to WT.

Conclusion: Although the p.His6Tyr and p.Ser118Pro variant receptors have conserved estrogen signaling, our preliminary data show that they decrease ERα expression in vitro. This could be an underlying pathophysiologic mechanism affecting some women with unexplained infertility. Additional in vitro and in vivo analyses can further characterize their pathogenicity.

Presentation: Friday, June 16, 2023
FRI438 A Unique Case Of Secondary Hypogonadism And The Covid Booster Vaccine
Authors: Syed Omar Zia , Shakir Muhammad , Sajid Khadijah , Sood Aayushi , Bansal Nidhi
Published: October 2023

Disclosure: O.Z. Syed: None. M. Shakir: None. K. Sajid: None. A. Sood: None. N. Bansal: None.

38-year-old male with no past medical or surgical history presented for an evaluation of his low testosterone. The patient transports patients in the hospital, he denies any medication use, including steroids, opioids, and antibiotics. He has a biological 8-year-old son and denied any history of viral infection recently and denied any history of testicular trauma, mumps, or radiation therapy. He started having fatigue, chronic mild headache, and generalized weakness after receiving his third dose of covid vaccine in January, however, he denied any diplopia, galactorrhea, hirsutism, or sexual dysfunction. He did not report any side effects with the previous 2 covid vaccines. After a couple of months of persistent symptoms accompanied by 40 pounds of weight loss( half of which he regained in a few months without intervention), he went to his PCP for evaluation. On physical examination, there were no abnormalities seen and as per the patient, there was no change in testicular size either. The patient was found to have mildly elevated prolactin of 17.8 with low free and total testosterone of 134 accompanied by FSH of less than 1.0 and LH of 1.4. The remaining labs include but are not limited to CBC, CMP, TSH, Vitamin D, CK, Sex hormone-binding globulin, and cortisol. The autoimmune panel, Hepatitis, HIV, and tick panels were negative. While he was being worked up he tested himself multiple times for covid and was negative. The patient had an MRI done which did not show any mass or lesions. Furthermore, he also had a Chest x-ray and CT of his abdomen/pelvis along with an Ultrasound of the abdomen, which did not show any concerning findings. The patient was started on testosterone gel at the endocrinology clinic with regular follow-ups to assess for response and side effects.As per the currently available literature, there have been multiple associations made between Covid 19 infection and hypogonadism. There have been reports of low testosterone following covid 19 infections and conversely there have been worse outcomes of covid 19 related to men with lower levels of testosterone. This could possibly be due to ACE 2 expression on testicular cells or due to secondary inflammation and oxidative stress inhbitiing steroidogenesis. However no literature was found linking the covid 19 vaccine to secondary hypogonadim other than a case of pituitary apoplexy. These findings in light of our case warrant further research into possible association between the covid vaccine and hypogonadism. In patients presenting with generalized weakness and similar vague symptoms, we need to consider a history of covid vaccination as it could potentially be a cause of secondary hypogonadism. Furthermore, more detailed and larger-scale studies need to be done concerning the covid booster vaccines and hypogonadism.

Presentation: Friday, June 16, 2023
FRI439 Spermatogenesis Induction For Hypogonadotrophic Hypogonadism In An Australian Tertiary Hospital Andrology Service
Authors: Sarlos Stella , Herath Madhuni , Sim Ie-Wen , Upreti Rita , McLachlan Robert Ian , Allan Carolyn A
Published: October 2023

Disclosure: S. Sarlos: None. M. Herath: None. I. Sim: None. R. Upreti: None. R.I. McLachlan: None. C.A. Allan: None.

Infertility affects 8-12% of couples with male factors contributing to 50% and solely responsible in 20-30% of cases. Male infertility due to hypogonadotrophic hypogonadism (HH) is amenable to medical treatment. We report a series of 23 men treated for infertility from HH (congenital n=13, acquired n=10) with a median age of 33 years. Causes included panhypopituitarism (n=11) [empty sella (n=2), adenoma (n=3), craniopharyngioma (n=1), pituitary hypoplasia (n=3), CHARGE syndrome (n=1), cranial radiotherapy (n=1)], Kallmann syndrome (n=1), thalassaemia major (n=1) and idiopathic (n=10). Urinary or recombinant human chorionic gonadotrophin (hCG) as an LH substitute was used based on established protocols. Commencing doses were 1500 IU or 62.5mcg s/cut twice weekly respectively and titrated to achieve normal serum total testosterone. Recombinant FSH was added after 6 months for persisting azoospermia. Wherever possible, men previously treated with long-acting injectable testosterone undecanoate were transitioned to transdermal testosterone for a minimum of 6 months prior to initiation of gonadotrophin therapy. There was wide variability in time to first appearance of sperm (median 16 months, range 4 - 42). In androgen treatment-naive men with acquired HH in adulthood (n=9), sperm was detected at a median 8 months (range 3 - 21 months) and for 4 men, hCG monotherapy was sufficient. For men with congenital causes of HH (n=13), time to first sperm detection was longer (median 17 months, range 12 - 42) and all required addition of FSH. In men previously treated with intramuscular testosterone (n=8), the median time to sperm appearance was 17 months (range 12-24) as compared to 13 months (5-42) in men treated with transdermal testosterone (n=6). To date, 11 of the 16 men actively seeking fertility have had children, with 7 requiring assisted reproduction. Gonadotrophin therapy is an effective fertility treatment for most men with HH. Clinicians can anticipate a longer time to spermatogenesis in men pre-treated with intramuscular androgens and those with congenital HH. Couples often move to assisted reproductive therapy, likely due to considerations such as semen quality, co-existing female factors and/or desire for prompt conception.

Presentation: Friday, June 16, 2023
FRI454 The Effect Of Growth Hormone Treatment According To Karyotype Classification In Turner Syndrome Patients
Authors: Choi Jung Eun , Yu Jeesuk , Kim Hae Soon
Published: October 2023

Disclosure: J. Choi: None. J. Yu: None. H. Kim: None.

OBJECTIVES Short stature is the main characteristics for Turner syndrome (TS) patients, and growth hormone (GH) therapy has been used as an essential treatment for developing final adult height. However, there are only a few studies on the difference responsiveness to GH therapy according to the karyotype of Turner syndrome in Korea. The aim of this study was to analyze the effect of different types of TS karyotype abnormality on the response of GH therapy. METHODS The clinical parameters of 194 TS patients registered in the LG Growth Study, who received rhGH therapy (Eutropin inj.,LG Chem Ltd.,Korea) between February 2001 and October 2021, were retrospectively reviewed. Data for four group of subjects were obtained as follow: Monosomy X (n=56); Mosaicism without X structural abnormality (n=41); Mosaicism with X structural abnormality (n=71); X Structural abnormality (n=26). Parameters of biochemical characteristics and growth responses were compared during 3-year GH treatment. RESULTS The baseline height standard deviation score (SDS) was -2.85±0.86 in total TS patients. The growth velocity (cm/year) in the second year was higher in the Mosaicism without X structural abnormality compared to the monosomy X patients. However, the change of Hight SDS over three years was not statistically different between karyotypes. The growth rate was the highest in the first year, and it showed a decreasing trend over time. There was an increase in Height SDS due to growth hormone treatment for three years. The variables affecting the 0-3rd Δ height SDS were not found in the regression analysis CONCLUSIONS Our results indicate that there are no difference in response to three years of growth hormone therapy according to karyotype of TS. It was found that the initial height SDS was the lowest in monosomy compared to the others group. Further studies should be performed on other factors influencing the efficacy of the therapy.

Presentation Date: Friday, June 16, 2023
SAT361 A Rare Case of Ovarian Steroid Cell Tumor in a Patient With Known Polycystic Ovarian Syndrome
Authors: Aluko Oluwatomisin A , Patel Shivani , Graul Ashley , Deshmukh Mrunalini
Published: October 2023

Disclosure: O.A. Aluko: None. S. Patel: None. A. Graul: None. M. Deshmukh: None.

Introduction.Sex cord stromal cell tumors are rare tumors of the ovary that usually occur inthe first twotothreedecades of life. Specifically, steroid cell tumors of the ovary account for 0.1% of all ovariantumors.Testosterone productionis typical, with hirsutism and virilizationbeing the most commonsymptoms. Amarkedly elevated testosterone level >200ng/dl is animportant diagnostic threshold for discriminatingandrogen- secreting tumors from non-neoplastic lesions.We report acase of ovariansteroid cell tumor inapatient with knownPCOS.Case ReportA 28-year old female with past medical history of PCOS and infertility was referred tothe Endocrinologyoffice for evaluationof worsening hirsutism and elevated testosterone level.She reported increased facial hair especially onher upper lipand chin, abdomenand inner thighs. Shehad ahistory of irregular periods and had beenonoral contraceptives as aresult. She was started onspironolactone 50mg daily by her PCP with noimprovement insymptoms.Bloodwork done prior tovisit showed elevated total testosterone of 312ng/dL (n13-71ng/dL) withelevated free testosterone of 5.8pg/dL (n0.0- 4.2pg/dL). Pelvic ultrasound showed peripherally orientedfollicles inkeeping with knowndiagnosis of PCOS with nosuspicious adnexal mass or loculatedcollections. Repeat bloodwork done three weeks after discontinuing spironolactone showed worsening elevatedserum testosterone of 561ng/dL with elevated free testosterone of 7.7pg/dL. Further workuptorule outother underlying causes returned negative. DHEA-S, Androstenedione, 24hr urinary cortisol, salivarycortisol X3, ACTH, InhibinB, LDH, Ca-125, betaHCG, were all withinnormal limits.CT abdomenand pelvis showed anenhancing nodule inthe right ovary with normal adrenal glands.Repeat pelvic ultrasound showed anew 2.9cm right ovariansolid isoechoic hyperemic nodule O-RADScategory 5(50% risk of malignancy) similar toMRI findings.She underwent aright laparoscopic ovariancystectomy with pathology diagnosis of sex cord steroidal celltumor with noevidence of increased mitotic activity or necrosis. Immuno-histochemistry supported thediagnosis with positive steroidogenic factor-1(SF-1) gene. Repeat total testosterone and freetestosterone levels six weeks after surgery were 44ng/dl and 2.4ng/dl respectively, both withinnormallimits.ConclusionInthe setting of rapid onset hirsutism with significantly elevated testosterone levels, there should be ahigh clinical suspicionfor anandrogensecreting tumor which must be excluded, irrespective of knowndiagnosis of PCOS. Repeat imaging after non-neoplastic processes are ruled out should be consideredas surgical resectionis the primary treatment for ovariansteroid cell tumors and should not be missed.

Presentation Date: Saturday, June 17, 2023
SAT446 Association of Polychlorinated Biphenyls With the Hormonal Parameters of the Normal Menstrual Cycle May Contribute to Unexplained Infertility
Authors: Butler Alexandra E , Brennan Edwina , Sathyapalan Thozhukat , Lawrence Atkin Stephen
Published: October 2023

Disclosure: A.E. Butler: None. E. Brennan: None. T. Sathyapalan: None. S.L. Atkin: None.

Introduction: Polychlorinated biphenyls (PCBs) are organic lipophilic pollutants that accumulate through the diet and increase with age and obesity. Previous research has linked PCBs with menstrual function; therefore, this study was undertaken to look at the association of PCBs with parameters of menstrual function at day 21 of the menstrual cycle in a group of women who were undergoing in vitro fertilization (IVF). Methods: 29 Caucasian women from a UK academic center aged 32.9±4.4 years, with body mass index BMI 25.4±3.6 kg/m2, who were undergoing in vitro fertilization were recruited. Five of the women had unexplained infertility and the remainder (n=24) had male factor infertility or tubal problems. Blood was withdrawn at day 21 of their menstrual cycle, at the time of mock embryo transfer. PCBs were measured using high resolution gas chromatography. Results: Only PCB118, PCB153, PCB138 and PCB180 were detected. PCB153, PCB138, PCB180 inversely correlated with estradiol (p<0.05). PCB 118 and PCB138 inversely correlated with FSH (p<0.05). PCB118, PCB153, PCB138 and PCB180 inversely correlated with LH (p<0.05, p<0.01, p<0.01 and p<0.01, respectively). No correlation was seen between PCBs and progesterone. Women with unexplained infertility showed higher levels of PCB118, PCB153, PCB138 and PCB180 (p<0.05) compared to those with male factor infertility though other hormonal parameters did not differ other than FSH that was lower in the unexplained group (p=0.01). Conclusion: These data are in accord with an effect of PCBs on the menstrual cycle in normal women and may be involved in the mechanism of unexplained infertility.

Presentation Date: Saturday, June 17, 2023
OR35-04 Anti-Müllerian Hormone Variants identified in Patients with Polycystic Ovary Syndrome Affect Secretion and Immunoactivity
Authors: Meng Li , McLuskey Anke , Elizabeth Dunaif Andrea , Visser Jenny A
Published: October 2023

Disclosure: L. Meng: None. A. McLuskey: None. A.E. Dunaif: None. J.A. Visser: None.

Recently rare heterozygous AMH variants were identified in women with PCOS, causing reduced AMH signaling. However, the exact functional mechanism remains unknown. Here, we analyzed the processing, secretion and signaling of these AMH variants. Six PCOS-specific AMH variants (V12G, P151S, P270S, P352S, P362S, H506Q) and one control-specific variant (A519V) were introduced in an AMH expression vector. Signaling activity was measured using the BRE-Luc reporter in the mouse granulosa cell-line KK-1. AMH cleavage and secretion was analyzed by Western Blot, confocal microscopy and ELISA. Serum AMH levels in PCOS cases harboring these variants were measured by ELISA. hAMH-151S and hAMH-506Q have ∼90% decreased AMH signaling (P<0.001). The other five variants were comparable to wild-type (wt)-AMH signaling. Coexpression of hAMH-151S or hAMH-506Q with wt-hAMH at equal amounts inhibited wt-hAMH signaling by ∼30% (P<0.001). Western blot analysis revealed that hAMH-151S and hAMH-506Q proteins were only detected in the cell lysate but not in the supernatant, while wt-hAMH and the hAMH-352S and hAMH-362S variants were detected in both the cell lysate and the supernatant. Confocal analysis showed that cells expressing hAMH-151S and hAMH-506Q had higher cellular AMH protein levels with ER retention compared to cells expressing wt-hAMH and hAMH-352S. In agreement, using two commercial AMH ELISA’s, hAMH-151S was readily detected in the cell lysate, while only very low levels were detected in the supernatant. Both hAMH-362S and hAMH-519V were detectable using the Fujirebio-AMH ELISA, but showed severely reduced immunoactivity measured by the pico-AMH ELISA. Surprisingly, hAMH-506Q was undetectable in both the cell lysate and supernatant by both ELISA’s. However, in PCOS cases, heterozygous carriers of the P151S and H506Q variants still had detectable AMH in both assays. In conclusion, our results show that P151S and H506Q disrupt normal processing and secretion of AMH, leading to hampered secretion of wt-AMH. Additionally, AMH variants impair the AMH immunoactivity with (H506Q) or without (P362S and A519V) influencing their bioactivities.

Presentation Date: Sunday, June 18, 2023
THU573 Paxillin Regulates Androgen Receptor In Granulosa Cells
Authors: Weidner Adelaide , Roy Anna , Candidate B S , Vann Kenji , Astapova Olga
Published: October 2023

Disclosure: A. Weidner: None. A. Roy: None. K. Vann: None. O. Astapova: None.

Paxillin is a ubiquitously-expressed adaptor protein that is integral in focal adhesions and plays an active role in the regulation of cell motility and apoptosis. Recently, paxillin was shown to mediate pro-tumorigenic androgen signaling in prostate cancer cells, where it is necessary for rapid cytoplasmic kinase signaling triggered by activation of membrane-bound androgen receptor. We sought to investigate the relationship between paxillin and androgen receptor (AR) in granulosa cells, where androgen actions, apoptosis and focal adhesions are of known importance, but the role of paxillin is understudied. When paxillin expression was silenced using siRNA in human granulosa-derived KGN cells, AR protein expression was significantly reduced, while its RNA expression was unaffected. Similarly, in granulosa cell-specific paxillin knockout mice, AR protein expression in granulosa cells was reduced. Further, we found that AR and paxillin co-localize in focal adhesions in mouse primary granulosa cells and in KGN cells, and their interaction requires the activity of focal adhesion kinase. Disruption of focal adhesion with a focal adhesion kinase inhibitor also leads to reduced AR protein levels. To further explore the mechanisms by which paxillin regulates AR expression, we created paxillin-null KGN cell lines using CRISPR-Cas9 gene editing. While paxillin loss resulted in reduced AR protein expression early (days) in the gene-editing process, this effect disappeared after weeks of clonal expansion from selected single cells. Expanded paxillin-null KGN clones showed normal AR protein expression, but increased AR mRNA expression, suggesting an adaptive increase in AR gene transcription to compensate for reduced AR protein levels in cells that survived paxillin deletion. Studies of AR protein degradation using cycloheximide to block protein synthesis showed that the half-life of AR was reduced from 6.7 to 4.5 hours (approximately 33%) by paxillin deletion, indicating accelerated AR protein degradation in cells lacking paxillin. Based on our results, we hypothesize that paxillin brings AR into focal adhesions in granulosa cells, thereby sequestering it from proteasomal degradation in the cytoplasm. Thus, paxillin may be a novel target in the management of androgen-related disorders in women, such as polycystic ovary syndrome.

Presentation Date: Thursday, June 15, 2023
SAT013 Characterization Of CRISPR/Cas9 Induced RXR Isotype Mutants In Xenopus Laevis
Authors: Furlow J David , Saini Simran , Mengeling Brenda J , Horb Marko E
Published: October 2023

Disclosure: J.D. Furlow: None. S. Saini: None. B.J. Mengeling: None. M.E. Horb: None.

Retinoid-X receptors (RXRs) occupy a unique position in the nuclear receptor (NR) superfamily. While some actions of the highly conserved RXRs may be independent, RXRs most often are heterodimer partners for several NRs that regulate development, reproduction and metabolism. As such, environmental chemicals interfering with the activity of RXRs and their partners may have significant deleterious effects on human health, often in developmental window, cell-type, and gene selective ways. The frog Xenopus has long served as an excellent model for studying vertebrate development including thyroid hormone action, and recent advances in genome editing have greatly enhanced its utility for investigating basic cell and organismal biology as well as human disease processes. To determine isotype specific roles in response to natural and synthetic ligands as well as candidate environmental chemicals interacting with RXRs in vivo, we created targeted germline mutations in the Xenopus tropicalis RXRα (rxra), RXRβ (rxrb), and RXRγ (rxrg) genes. For CRISPR/Cas9 mediated genome editing, guide RNAs were designed to target sequences just prior to or within the first RXR DNA binding domains. We raised all three RXR homozygous null mutants up to and through metamorphosis. The rxrb and rxrg homozygous null mutants completed metamorphosis and allowed us to establish adult null lines, but the rxra homozygous null mutants die at metamorphic climax, reminiscent of the thyroid hormone receptor (thra/thrb) double mutants. Some rxra mutant froglets also lack forearms or have forelimb defects, implying a role for rxra in RA mediated forelimb specification. The X. tropicalis rxrg homozygous null mutants show a trend toward an increased rate of metamorphosis versus matched wild-type controls, with significantly elevated tshb gene expression as newly metamorphosed froglets suggesting impaired negative feedback by TH. Significantly altered opsin gene expression in the froglet eye was also observed in rxrg mutant retinas: elevated short wave opsins opn1sw1 and opn1sw2 and sharply reduced long wave opsin opn1lw. This elevated tshb and opn1sw1 expression is highly similar to what is found in mouse rxrg null mutants; thus RXRγ action in the eye and pituitary appears to be highly conserved between mammals and amphibians. The establishment of these new RXR isotype mutant specific lines will support the expanded use of Xenopus to study basic biology and environmental toxicology involving TR-RXR and other NR-RXR partnerships.

Presentation: Saturday, June 17, 2023
SAT015 Improving The Quality Of Disease Biomarker Measurements In Patient Care And Research-CDC's Clinical Standardization Programs
Authors: Sugahara Otoe , Buchannan Tatiana , Danilenko Uliana , Ribera Ashley , Collins Lynn , Vasquez Norma , Dahya Komal , Pokuah Fidelia , Zhou Hui , Zhang Li , Lyle Alicia N , Vesper Hubert W , Long Lindsey , Knoblock Samantha , Flores Elias , Doty Alex
Published: October 2023

Disclosure: O. Sugahara: None. T. Buchannan: None. U. Danilenko: None. A. Ribera: None. L. Collins: None. N. Vasquez: None. K. Dahya: None. F. Pokuah: None. H. Zhou: None. L. Zhang: None. A.N. Lyle: None. H.W. Vesper: None.

High-quality disease biomarker tests are critical for the correct diagnosis and treatment of patients, accurate interpretation of research data, and effective use of research findings in health care. CDC’s Clinical Standardization Programs (CDC CSP) assist researchers and laboratories with assessing and improving the analytical performance of biomarker tests performed in patient care, public health, and research. CDC CSP develops and applies reference methods, materials, and protocols to consistently calibrate biomarker tests across instruments and laboratories and to assess and improve other analytical performance characteristics, such as analytical specificity. CDC CSP works with test manufacturers and laboratories operating in-house developed tests to correctly calibrate tests. Additional programs help clinical and research laboratories monitor the accuracy and reliability of tests over time. Furthermore, CDC CSP collaborates with researchers on the development of reference intervals and on using new and emerging biomarkers in patient care and public health. CDC CSP provides comprehensive programs for traditional blood lipids, testosterone, estradiol, vitamin D, and free thyroxine. Programs for free testosterone, parathyroid hormones, apolipoproteins, and angiotensin peptides will be available soon. CDC CSP collaborated with members of the Partnership for the Accurate Testing of Hormones (PATH) and established reference intervals for testosterone in men. CDC CSP is currently working to develop reference intervals for testosterone in women and for estradiol in postmenopausal women. A CDC CSP interlaboratory study on free thyroxine measurements showed high variability in calibration accuracy among free thyroxine assays. CDC CSP is using its reference methods and materials to minimize this variability. It is anticipated that assay recalibration may change measurement values for some tests by up to 50%. CDC CSP is working with PATH and the International Federation for Clinical Chemistry and Laboratory Medicine (IFCC) to educate stakeholders about the upcoming change in free thyroxine measurements.

Presentation: Saturday, June 17, 2023
OR22-03 Enhanced Endothelial Mineralocorticoid Receptor Signaling Mediates Diet - Induced Soleus Fat Infiltration And Insulin Resistance
Authors: Homan Carlton , Habibi Javad , Chen Dongqing , Whaley-Connell Adam , Sowers James R , Jia Guanghong
Published: October 2023

Disclosure: C. Homan: None. J. Habibi: None. D. Chen: None. A. Whaley-Connell: None. J.R. Sowers: None. G. Jia: None.

Skeletal muscle is an important organ in the storage and release of lipids in response to physiological changes in free fatty acid (FFA) supply and demand. Once fat tissue mass expansion exceeds its blood supply excess lipids are stored ectopically as small lipid droplets in non-adipose tissues such as skeletal muscle. Recently, our data found that mineralocorticoid receptors (MRs) mediate Western diet (WD)-induced lipid infiltration of skeletal muscle and insulin resistance. Related to this, MRs, the principal receptors for the hormone aldosterone in the kidney, also exist in vascular cells, including endothelial cells (ECs). Moreover, enhanced MR signaling in ECs (ECMR) induces arterial stiffening and impairs microcirculation. However, our understanding of the precise mechanisms by which enhanced ECMR activation promotes skeletal muscle insulin resistance remains unclear. In this study we investigated the roles of ECMR in soleus lipid accumulation and corresponding insulin resistance in diet-induced obesity. Six-week-old ECMR wild type (ECMR+/+) and knockout (ECMR-/-) mice were fed either a mouse chow diet or WD for 16 weeks. 16 weeks of WD induced increases in glucose intolerance, fasting plasma insulin levels, HOMA-IR and insulin resistance index were found to be blunted in ECMR-/- mice. ECMR-/- prevented WD-induced increase in soleus FFA levels without changing serum FFA levels. Meanwhile, WD also increased soleus intramyocellular lipid content, plasma EC derived exosomal CD36 and soleus CD36 protein levels. These abnormalities were related to reduced soleus insulin metabolic signaling in PI3K/Akt pathways, as these pathways were blunted in ECMR-/- mice. These findings indicate that EC specific MR activation contributes to diet-induced increases in CD36 expression, soleus fat infiltration, as well as systemic and skeletal muscle insulin resistance.

Presentation: Saturday, June 17, 2023
OR33-02 The Effects Of Timing And Patterns Of Hormone Stimulation On Single Molecule Dynamics And Gene Regulation In Vivo
Authors: Stavreva Diana A , Hoang Le , Wagh Kaustubh , Chari Raj , Upadhyaya Arpita , Hager Gordon L
Published: October 2023

Disclosure: D.A. Stavreva: None. L. Hoang: None. K. Wagh: None. R. Chari: None. A. Upadhyaya: None. G.L. Hager: None.

Even though most studies on hormonal signaling are performed under continuous stimulation, secretion of many hormones follows a circadian pattern, comprised of periodic (ultradian) pulses with short duration. Using glucocorticoid receptor (GR) signaling as a model system we have previously demonstrated that receptor occupancy of regulatory elements precisely tracks the hormonal pulses resulting in cyclic GR-mediated transcription (gene pulsing) (Stavreva et al., NCB 2009). Using various genomic approaches, we have also uncovered the impact of ultradian and constant stimulation on the lifetimes of chromatin accessibility and long-range promoter/enhancer interactions (Stavreva et al., Genome Res. 2015). By visualizing RNA synthesis from single-copy GR-regulated transcription sites (TSs) we demonstrated treatment-specific RNA bursting patterns. Moreover, our single molecule tracking (SMT) experiments showed that transcription factor (TF) dynamics determine burst duration and frequency (Stavreva et al., 2019). To further study the in vivo dynamics of various TFs we tagged the endogenous GR as well as the estrogen, progesterone receptor (ER and PR, respectively), and the large Pol II subunit (Rpb1) with Halo tag in MCF7 cells. We use SMT in combination with other microscopy and biochemical approaches to i) map the dynamics of TFs in response to constant and ultradian hormone stimulations, and ii) determine their effects on transcription regulation. These studies will help us better understand the importance of timing and pattern of hormone treatment for proper gene regulation in different physiological contexts.

Presentation: Sunday, June 18, 2023
THU674 Papillary Thyroid Carcinoma Presenting As A Benign Ectopic Thyroid Mass In The Anterior Mediastinum. A Case Report
Authors: Valdecanas David-Santos Mary Lourdes Grace , Narvacan-Montano Carolyn
Published: October 2023

Disclosure: M.V. David-santos: None. C. Narvacan-montano: None.

Background: Ectopic Thyroid Tissue (ETT) is a rare developmental anomaly that is due to the failure of the thyroid gland to assume its position in front of the trachea. Ectopic Thyroid tissue in the mediastinum is very rare, with only a few cases reported in the literature. An incidental Papillary Thyroid Carcinoma presenting as a mediastinal mass that turns out to be benign ectopic thyroid cells is even rarer. Case: This is the case of a 33-year-old female who presented with sudden onset difficulty of breathing and chest heaviness. On workup, she was found to have an anterior mediastinal mass, which showed ectopic thyroid cells on biopsy. Further evaluation of the neck area revealed a 1.25 cm x 1 x 1.13 cm nodule on the thyroid gland which showed suspicious for Papillary Thyroid Carcinoma on Fine needle aspiration biopsy. The patient underwent Total Thyroidectomy and Excision of the Anterior Mediastinal Mass. The histopathologic report of the thyroid mass revealed a Conventional Unifocal Papillary Thyroid Carcinoma, while the excised tissues comprising the mediastinal mass were consistent with benign ectopic thyroid tissues with colloid goiter. Conclusion: The presence of the ectopic thyroid mass in the anterior mediastinum may be a potential area for metastasis. The final pathologic report nevertheless, categorizes this Papillary thyroid carcinoma as having a low risk of metastasis as per the American Thyroid Association Classification of 2015. Hence the conservative approach of Suppression therapy and active surveillance is the choice of management for this unusual case.

Presentation: Thursday, June 15, 2023
FRI466 Toxic Retrosternal Goiter Following SARS-CoV-2 Vaccination
Authors: Schipper Joslyn , Abdul Sater Zahraa
Published: October 2023

Disclosure: J. Schipper: None. Z. Abdul Sater: None.

Since the outbreak of the SARS-CoV-2 pandemic, there have been reports of autoimmune thyroid diseases related to COVID inflection and SARS-CoV-2 vaccination. We report a case of SARS-CoV-2 mRNA vaccination (mCoVvax)-induced Graves with retrosternal extension. A 21-year-old Hispanic male with no previous history of thyroid illness presented with a 4-month history of tachycardia, tremors, agitation, unintentional weight loss, night sweats, low-grade fever, and orthopnea that developed 62 days after receiving the first dose of mCoVvax. His physical examination was significant for sinus tachycardia, hypertension, a resting tremor, brisk deep tendon reflexes, and diffuse nonpruritic maculopapular rash. His thyroid gland was diffusely enlarged with an unpalpable lower margin and positive Pemberton sign. TSH was undetectable with elevated Free T4 and Total T3 [11.0 (NL 0.8-1.8 ng/dL) and 800 (76-181 ng/dL), respectively]. Thyroid-stimulating immunoglobulin (TSI) was 337% (NL<140%), and thyroid peroxidase antibody was 653 IU/mL (NL < 9 IU/mL). Thyroid ultrasound showed a diffusely enlarged thyroid gland with diffusely increased thyroid uptake on technetium thyroid scan (95% and 85% uptake after 6 and 24 hrs, respectively), consistent with Graves’ disease. CT scan of the neck and chest confirmed retrosternal Goiter without tracheal deviation. He was treated with high-dose of Methimazole, Atenolol, and Prednisone. A literature search revealed five cases of Graves’ disease after mCOVvax, all of which did not have a retrosternal extension of the thyroid. Our case represents a rare complication of mCoVvax. Clinicians should consider retrosternal goiter in patients with thyrotoxicosis and orthopnea after mCoVvax exposure.

Presentation: Friday, June 16, 2023
FRI475 The Impact Of Suppression Of Androgen Signaling On Thyroid Hormone Homeostasis In Prostate Cancer Patients
Authors: David Karel , Devos Gaetan , Devlies Wout , Narinx Nick , Antonio Leen , Deboel Ludo , Schollaert Dieter , Vanderschueren Dirk M , Claessens Frank A , Joniau Steven , Decallonne Brigitte
Published: October 2023

Disclosure: K. David: None. G. Devos: None. W. Devlies: None. N. Narinx: None. L. Antonio: None. L. Deboel: None. D. Schollaert: None. D.M. Vanderschueren: None. F.A. Claessens: None. S. Joniau: None. B. Decallonne: None.

Next to the classical androgen deprivation therapy (ADT) with GnRH-(ant)agonists, androgen receptor signaling inhibitors (ARSIs) such as apalutamide (APA) have become available for the treatment of prostate cancer (PC) patients in the last years. Hypothyroidism was more frequent in APA-treated patients compared to placebo in the SPARTAN trial (NCT01946204) (8.1% vs 2%). We collected serum of PC patients included in a RCT (ARNEO; NCT03080116) before and after 12 weeks of neo-adjuvant ADT (GnRH-antagonist degarelix (DGX) with APA or placebo). We aimed to explore changes in thyroid homeostasis under deep androgen suppression with combined GnRH-antagonist and ARSI. Of the 88 patients ending the trial, 1 was excluded because of history of hypothyroidism for which he received levothyroxine. 43 and 44 patients were included in the DGX+placebo and DGX+APA group, respectively. Mean age was 66.3y. Sex steroid levels measured by LC-MS/MS decreased significantly to castrate range with median testosterone 10.5 ng/dL and estradiol 3.6 ng/L. TSH levels increased in both treatment groups, but more pronounced in the DGX+APA group (DGX+placebo before: 1.18 mIU/L and after: 1.33 mIU/L, p 0.0351 vs. DGX+APA before: 1.07 mIU/L and after: 1.87 mIU/L, p <0.0001; difference between groups after treatment: p <0.0001). Free T4 and total T4 levels decreased in the DGX+APA group, but remained unaltered in the DGX+placebo group (free T4 after: 16.53 vs 13.87 pmol/L, p<0.0001 and total T4 after: 8.77 vs. 7.78 µg/dL, p 0.0055). Total T3 and free T3 increased in the DGX+APA group, and again did not differ after treatment in the DGX+placebo group. (free T3 after: 5.03 vs 5.41 pmol/L, p 0.0066 and total T3 after: 132.30 vs 149.50 ng/dL, p 0.0003). As a result, the total and free T3/T4 ratio increased in the DGX+APA group with 32% and 35%, respectively. Reverse T3 decreased in both treatment arms, but greater in the DGX+APA group (19.55 vs 14.20 ng/dL, p 0.0157). Thyroxine binding globulin (TBG) increased in both treatment arms, with 6.9% and 8.4%, respectively. In conclusion, interference with androgen signaling alters thyroid homeostasis in PC patients, and is most pronounced under deep androgen suppression with combined GnRH-antagonist and ARSI therapy. The increase in T3 and decrease in rT3 may indicate additional effects of ARSI on deiodinase activity. Longer follow-up trials are needed to confirm these findings and assess the clinical implications of these changes in thyroid homeostasis.

Presentation: Friday, June 16, 2023
FRI477 The Effect Of Gestational Selective Serotonin Reuptake Inhibitor (SSRI) Treatment On Newborn Thyroid Screen
Authors: Brener Avivit , Raviv Orian , Cohen-Sela Eyal , Yackobovitch-Gavan Michal , Almashanu Shlomo , Marom Ronella , Hiersch Liran , Lebenthal Yael
Published: October 2023

Disclosure: A. Brener: None. O. Raviv: None. E. Cohen-Sela: None. M. Yackobovitch-Gavan: None. S. Almashanu: None. R. Marom: None. L. Hiersch: None. Y. Lebenthal: None.

Background: Clinical depression treated by selective serotonin reuptake inhibitors (SSRIs) has become relatively common in women of childbearing age. The effect of gestational exposure to SSRIs on the newborn’s hypothalamic-pituitary-thyroid postnatal adaptation has not yet been determined. We aimed to investigate the characteristics of the mother–infant dyad of term infants of mothers treated with SSRIs for depression during pregnancy. Methods: This observational study included mother-infant dyads of infants delivered at term in a single tertiary medical center between 2011 and 2021. Data on maternal, pregnancy, delivery, and perinatal characteristics of the mother-infant dyads were retrieved from the hospital’s electronic database. The clinical data were linked with the newborn thyroid screening results of total thyroxin (TT4) obtained at 48-72 hours since delivery. Excluded from analysis were mothers with other chronic medical conditions and newborns with genetic syndromes. Primary outcome measures were newborn TT4 levels and fetal growth as expressed by neonatal birthweight z-scores (WHO charts). Results: SSRI anti-depression treatment during gestation was reported in 2577 (2.4%) out of 106,173 infant-mother dyads. Mothers treated with SSRIs were older (35.0±4.8 vs 32.6±4.8 years, p<0.001), had a higher pre-pregnancy body mass index (23.4±4.5 vs 22.7±4.1, p<0.001) and a similar mean weight gain of 13 kg during pregnancy. Elective and emergent cesarean deliveries were more common among SSRI-treated mothers (14.8% vs 11.1% and 16.5% vs 8.6%, respectively, p<0.001). Infants of those mothers were born earlier (gestational age [GA] 38.7±1.9 vs 39.5±11.5 weeks, p<0.001), had lower birthweight z-scores (-0.48±1.24 vs -0.04±0.92, p<0.001) and lower TT4 levels (14.7±3.9 vs 15.0±3.7, p=0.001). A multivariate linear regression model revealed that sex, GA and birthweight z-score were highly significant contributors to TT4 levels (p<0.001 for all), while SSRI treatment during pregnancy was not (p=0.497). Conclusions: Our findings suggest that anti-depressant use of SSRIs during pregnancy does not have a direct effect on the postnatal adaptation of the hypothalamic-pituitary-thyroidal axis. However, differences in fetal growth and in characteristics of delivery may determine newborn thyroid function.

Presentation: Friday, June 16, 2023
FRI480 Uncovering The Mystery Of Macro TSH
Authors: Anwar Farrah , Saeed Zeb I
Published: October 2023

Disclosure: F. Anwar: None. Z.I. Saeed: None.

Introduction: An isolated elevation in TSH level, while uncommon, may result in incorrect diagnosis and improper treatment. We present a case of suspected hypothyroidism that was later found to be caused by macroTSH in the blood. Clinical Case: A 21-year-old asymptomatic female was diagnosed with hypothyroidism and started on levothyroxine at age 16 when routine lab work revealed TSH of 34.2 MCU (0.550-4.780 MCU) and free T4 of 1.1 ng/dl (0.89-1.76 ng/dl). She denied any personal or family history of thyroid disorders in the past. Patient later reported symptoms including palpitations and weight loss leading to discontinuation of levothyroxine. Repeat lab work showed TSH of 0.112 MCU and free T4 of 1.71 ng/dl. After discontinuing levothyroxine, TSH levels were found in range of 89.26 MCU to 106 MCU (0.550-4.780 MCU). Free T4 levels remained normal, even when tested using dialysis method. Total T3 level also remained normal at 87 ng/dl (80-200 ng/dl). TPO antibodies were slightly elevated at 9.3 IU/mL (0.0-9.0 IU/ml), TSI and TSH receptor antibodies were negative. Patient's high TSH level, lack of symptoms, and normal free T4 levels prompted the use of different assays to test TSH. We contacted Mayo Clinic Lab for further testing. One assay (Roche Elecsys) showed TSH level of 90.8 mIU/L, but this result was not significantly changed when sample was incubated with a commercial heterophile antibody blocking reagent. Another assay (Beckman Coulter Access TSH) produced a significantly different but still elevated result of 43.1 mIU/L. For research purposes, the sample was further tested for macroTSH using polyethylene glycol (PEG) precipitation, which resulted in TSH level of 5.8 mIU/L which was reduced from 90.8 mIU/L (94% reduction). These results were higher than the mean +/- 2 standard deviations of the control group (range 0% to 65% reduction in TSH with PEG in controls). This suggested presence of macroTSH, although the assay's performance characteristics and reference ranges had not been established at the time of testing for this patient.

Patient's TSH levels remain high and she was advised not to begin the medication unless circulating thyroid hormone levels are abnormal. Conclusion: Macro TSH is a combination of monomeric TSH and anti-TSH antibodies, similar to macro-prolactin. It is considered inactive and has an estimated prevalence of 0.6-1.6%. In a review of over 150 patients, more than 50% of documented thyroid interferences, including macro TSH, led to incorrect diagnosis and improper treatment. This case demonstrates the significance of identifying macro TSH to avoid misdiagnosis and inappropriate management of euthyroid patients with isolated TSH elevation.

Presentation: Friday, June 16, 2023
FRI487 Oral Liquid L- Thyroxine (L-T4) Is More Effective In Controlling Hypothyroidism, Than L-T4 Tablet, In Patients Submitted To Total Thyroidectomy
Authors: Fallahi Poupak , Martina Ferrari Silvia , Ragusa Francesca , Elia Giusy , Materazzi Gabriele , Rosaria Paparo Sabrina , Patrizio Armando , Mazzi Valeria , Miccoli Paolo , Antonelli Alessandro
Published: October 2023

Disclosure: P. Fallahi: None. S. Ferrari: None. F. Ragusa: None. G. Elia: None. G. Materazzi: None. S. Paparo: None. A. Patrizio: None. V. Mazzi: None. P. Miccoli: None. A. Antonelli: None.

Objectives: Our aim is to investigate the efficacy of levothyroxine (L-T4) in liquid formulation in patients recently subjected to total thyroidectomy (without malabsorption or drug interference), with respect to L-T4 tablets. Methods: We enrolled two hundred and twenty-five patients of whom 75 received L-T4 in tablets, and 150 were treated with liquid L-T4 at the same dosage (1.5 mcg/kg/day). The therapy started the day after thyroidectomy, and the drugs were assumed every day 30 min before breakfast. The thyroid profile was assessed by evaluating serum thyrotropic hormone (TSH), free thyroxine (FT4), and free triiodothyronine (FT3) in a first control after 6 weeks, then in a second control after 12 weeks. Results: Our results showed that the prevalence of patients in the hypothyroid range (TSH>3.6 mcU/ml) was significantly higher in the L-T4 tablet group. Moreover, TSH values were significantly lower in the liquid L-T4 group, than in the tablet L-T4 group both at the first (P < 0.05), and at the second control (P < 0.01), while FT4 and FT3 levels were not significantly different. Conclusions: Our data showes that thyroidectomized patients, not reporting issues of malabsorption, gastric disorders, or drug interference, have a better control of TSH levels under a liquid L-T4 therapy.

Presentation: Friday, June 16, 2023