bluuebunny/crossref_metadata_2025_split huggingface dataset:
7277 Diabetic Ketoacidosis As The First Manifestation Of Ectopic Cushing’s Syndrome
Authors: Saudade Medeiros Regina , Pereira Bernardo , Pataco Andreia , Senra Catarina , João Ponte Maria , Anselmo João , Sousa Isabel 
Published: October 2024

Abstract
Disclosure: R.S. Medeiros: None.
Introduction: Ectopic Cushing’s Syndrome (ECS) represents 10-20% of all ACTH-dependent CS, and the majority of cases arise from lung carcinoids. These indolent lesions usually evolve clinically over 6-24 months, whereas carcinomas are associated with faster onset of ECS. Diabetic Ketoacidosis (DKA) as a presenting manifestation of Cushing’s Syndrome (CS) is very rare, with the few cases published to date related to Cushing’s disease. Clinical Case: A 42-year-old woman was admitted in the emergency room with DKA and COVID-19. During stabilization, florid CS was clinically suspected. She reported a 6-month history of secondary amenorrhea, lack of concentration and memory, easy bruising, proximal myopathy with frequent falls evolving to daily life dependency from relatives. She also reported a 2-month diagnosis of dyslipidemia, Diabetes and hypertension, and a diagnosis of atypical pneumonia 6 months ago. Physical exam showed typical cushingoid appearance, especially for marked limb sarcopenia and proximal myopathy. An overnight dexamethasone suppression test (morning cortisol: 25.7 ug/dL), 24h urinary free cortisol (UFC; 1072.5 ug/dL; reference: &lt;176) and serum cortisol at 23h00 (25.5 ug/dL) confirmed CS. ACTH-dependent CS was diagnosed. She had non-suppressible serum cortisol in high dose dexamethasone suppression test, and CRH stimulation test was suggestive of Cushing’s disease only by a 20% rise of cortisol at 30 minutes. Pituitary MRI was normal. Inferior Petrosal Sinus sampling was postponed for several months due to healthcare strikes. The patient started 750 mg of metyrapone in three divided doses but rapidly decreased the dose and stopped it due to clinical resolution of CS, a dramatic improvement of all glucocorticoid-related comorbidities leading to discontinuation of statin, insulin, main oral anti-hyperglycemic and antihypertensive medications, and normal midnight salivary cortisol and 24h UFC. She remained in this eucortisolemic state for 12 months. Thoracic CT scan revealed thymic hyperplasia and a 25x15 mm well-defined nodule in the lingula. A 68Ga-DOTANOC PET/CT showed a single uptake in the same lung region. The patient underwent lingulectomy plus excisional biopsy of interlobar lymph nodes, and pathology revealed a low-grade neuroendocrine tumor (Ki67&lt;2%, &lt;2 mitosis) without involved lymph nodes. The patient is weaning off from physiologic doses of hydrocortisone. Conclusion: DKA can rarely be the presenting manifestation of severe CS, including ECS. Clinicians should suspect on this etiology of DKA based on clinical signs and cumulative morbidity not typical of a young patient. This hint would lead to the expedite implementation of targeted therapies for initial stabilization of the CS patient, and the timely institution of a proper diagnostic approach and definitive treatment of this challenging patients.
Presentation: 6/2/2024
7232 Direct m6A mRNA-Seq In Endocrine- Resistant And Sensitive Breast Cancer Cells
Authors: Petri B J , Valdes B L , Piell K M , Rouchka E C , Klinge C M 
Published: October 2024

Abstract
Disclosure: B.J. Petri: None. B.L. Valdes: None. K.M. Piell: None. E.C. Rouchka: None. C.M. Klinge: None.
Acquired resistance to endocrine therapies (ET) in patients with estrogen receptor α (ER+) breast tumors results in metastatic spread. RNA modifications including N6-methyladenosine (m6A) play crucial roles in the post-transcriptional regulation of gene expression and have been implicated in cancer progression. We hypothesized that m6A epitranscriptomic alterations are associated with pathways in ET resistance. Direct-RNA sequencing technology (nanopore) was used to detect and map m6A modifications at single-nucleotide resolution to comprehensively profile m6A modifications in ET-resistant LCC9 and ET -sensitive MCF-7 breast cancer cell lines with or without 4-hydroxytamoxifen (4-OHT) treatment. Additionally, MCF-7 cells were treated with the METTL3 inhibitor STM2457 to identify m6A positions and transcripts directly regulated by METTL3 activity. We developed a new statistical method by integrating m6Anet, an existing machine-learning algorithm designed to call m6A modified bases, with a generalized linear model following a binomial distribution analysis to identify significant differential m6A modification ratios (DMR). We identified 61 transcripts with DMR between vehicle-treated LCC9 compared to MCF-7 cells and 323 transcripts with DMRs between 4-OHT-treated LCC9 and MCF-7 cells. Fewer transcripts showed DMR after 4-OHT treatment in MCF-7 (41) versus LCC9 cells (399). Thus, more changes in m6A DMRs were detected in response to 4-OHT in the ET-resistant LCC9 versus ET-sensitive MCF-7 cells with “translation elongation” as the top pathway in enrichment analysis. STM2457 treatment resulted in significant changes to DMRs in MCF-7 cells with or without 4-OHT treatment and altered gene expression patterns associated with “cytosolic ribosome” in enrichment analysis. We identified many genes with multiple m6A modifications, e.g., AMIGO2 with 14 m6A sites at METTL 3 target DRACH motifs in MCF-7 cells, compared to no m6A modification in AMIGO2 in LCC9 cells. Our findings reveal distinct m6A modification patterns in ET-resistant LCC9 breast cancer cells compared to their ET-sensitive parental MCF-7 cells and in response to 4-OHT, suggesting a potential role for epitranscriptomic alterations in the development of ET resistance.
Presentation: 6/3/2024
8767 Ectopic Cushing's Disease
Authors: Kothari V A , Yavuz S 
Published: October 2024

Abstract
Disclosure: V.A. Kothari: None.
Here we present the diagnostic journey of a patient with Cushing&apos;s syndrome attributed to a rare and elusive culprit—a neuroendocrine tumor that aberrantly produces adrenocorticotropic hormone (ACTH). An 83-year-old female with PMH of Graves&apos; disease well-controlled on methimazole 5 mg daily presented w/ AMS & worsening weakness and was found to have severe hypokalemia w/ K of 2.2. A CT chest done a month prior for bronchitis showed a right lower lobe lung nodule and a prior CT Abd also done a month prior showed an incidental pancreatic cyst of 1.5 cm and bilateral adrenal thickening. In addition, the patient had progressively worsening diabetes and proximal muscle loss. Patient was started on K supplementation and also spironolactone. A renin and aldosterone level were obtained which were 0.47 and &lt;1, respectively which ruled out hyperaldosteronism. Her insulin requirements had significantly escalated. Given the patient&apos;s skin changes with petechiae along with the findings above, the patient underwent a low-dose DST twice which showed elevated AM cortisol of 76.4 & 69.6 respectively. Dexamethasone level was appropriately elevated at 217. ACTH was 149. A 24 hr urine cortisol was repeated twice and showed 4477.6 and 7107.2 respectively. MRI brain showed a flatten pituitary and empty sella. Amiloride and ketoconazole were started. Spironolactone was increased to 200 mg. Ga-68 DOTATE PET/CT was done however did not show a distinct source lesion. A few weeks later,the patient again had to go to ER due to profound hyperkalemia and hyponatremia. Her medications were adjusted. While hospitalized, she underwent biopsy and cryoablation of the right lung nodule w/ subsequent immediate reduction in ACTH levels. However, although the ACTH level initially declined and a subsequent CT chest also showed the ablation zone to encompass the whole tumor, the patient again had worsening of ACTH & cortisol causing hypokalemia again. It was unclear if the patient&apos;s active Cushing&apos;s disease was due to residual tumor or other areas of carcinoid as the lung nodule was dotatate and FDG negative. Given the active Cushing&apos;s disease, ketoconazole 200 mg BID was restarted. Her condition continued to worsen despite escalating doses of ketoconazole. Due to proximal myopathy, she was unable to ambulate on her own requiring significant assistance. Definitive treatment was discussed with the patient and her family given the progressive functional decline. Broached adrenal ablation versus adrenalectomy was discussed and bilateral adrenalectomy was eventually decided upon and scheduled. This case highlights the rare occurrence of ectopic ACTH producing neuroendocrine tumors causing Cushing&apos;s syndrome. It underscores the need for advanced diagnostic approaches and collaborative healthcare efforts in managing uncommon endocrine disorders.
Presentation: 6/2/2024
7908 GPNMB Promotes Tumor Growth And Is A Biomarker For Lymphangioleiomyomatosis (LAM)
Authors: Gibbons Erin , Taya Manisha , Wu Huixing , Lopa Samia , Moss Joel , Henske Elizabeth P , McCormack Francis X , Hammes Stephen R 
Published: October 2024

Abstract
Disclosure: E. Gibbons: None. M. Taya: None. H. Wu: None. S. Lopa: None. J. Moss: None. E.P. Henske: None. F.X. McCormack: None. S.R. Hammes: None.
Lymphangioleiomyomatosis (LAM) is a rare, progressive cystic lung disease affecting almost exclusively female-sexed individuals. The cysts represent regions of lung destruction caused by smooth muscle tumors containing mutations in one of the two tuberous sclerosis (TSC) genes. mTORC1 inhibition slows but does not stop LAM advancement. Furthermore, monitoring disease progression is hindered by insufficient biomarkers. Therefore, new treatment options and biomarkers are needed. LAM cells express melanocytic markers, including Glycoprotein Non-Metastatic Melanoma Protein B (GPNMB). The function of GPNMB in LAM is currently unknown; however, GPNMB’s unique cell surface expression on tumor versus benign cells makes GPNMB a potential therapeutic target, and persistent release of its extracellular ectodomain suggests potential as a serum biomarker. Here we establish that GPNMB regulates TSC2-null tumor cell invasion as well as TSC2-null xenograft tumor growth, and that GPNMB expression is dependent on mTORC1 signaling. Further, we show that GPNMB’s ectodomain is released from the cell surface of TSC2-null cells by ADAM10 and 17 proteases, and we identify the protease target sequence on GPNMB. In fact, mutating this cleavage sequence so that the GPNMB ectodomain can no longer be shed results in cessation of TSC2-null tumor growth. Finally, we demonstrate that GPNMB’s ectodomain is present at higher levels in LAM patient serum compared to healthy controls, and that ectodomain levels decrease with mTORC1 inhibition, making it a potential LAM biomarker.
Presentation: 6/3/2024
8770 Highly Multiplexed Mapping Of The Ductal Carcinoma In Situ Ecosystem To Predict Disease Outcome
Authors: Wilson G M , Doan T B , Guild B J , Pathmanathan N , Graham J D 
Published: October 2024

Abstract
Disclosure: G.M. Wilson: None. T.B. Doan: None. B.J. Guild: None. N. Pathmanathan: None. J.D. Graham: None.
Mammographic screening programs have resulted in remarkable improvements in breast cancer survival due to early-stage detection. However, this has resulted in a dramatic increase in the detection of non-invasive ductal carcinoma in situ (DCIS), a non-obligate precursor of invasive ductal carcinoma (IDC), which is confined to the ducts and surrounded by a normal myoepithelial cell layer. Many DCIS cases will not progress to IDC within the lifetime of the patient. However, there are limited tools to accurately predict progression, thus virtually all DCIS is surgically removed, and most patients receive adjuvant radiotherapy as it halves recurrence rates. This likely represents overtreatment for a majority of patients. Emerging evidence has revealed that the adjacent microenvironment becomes progressively altered in DCIS and IDC compared to normal tissue, and this is correlated with disease outcome. The objective of this study was to investigate changes to the DCIS microenvironment during disease progression and identify disease states that predict patient outcome. We used immunohistochemistry and imaging mass cytometry with a well-annotated cohort of DCIS, with or without adjacent IDC, to measure a panel of 40 phenotypic and functional markers that allowed us to distinguish the diverse cell populations present in the tissues. We observed striking changes to almost all cell types within the DCIS tissue when comparing pure DCIS cases to DCIS adjacent to IDC. In particular, there were considerable changes to tissue-resident immune populations, with differences observed in the type, location and population size of a variety of immune cell subsets, in IDC-associated DCIS compared with DCIS-only cases. Computational mapping of cell type communities in pure and IDC-adjacent DCIS tissues revealed specific differences that could be used to predict the emergence of invasive disease, informing clinical management of this good prognosis malignancy and potentially avoiding over-treatment.
Presentation: 6/3/2024
8817 IGF-II Regulates Magmas A Mitochondrial Protein Required For Cell Viability And ROS Protection
Authors: Granados Katherine , Durán Alfonso , Almaguel Frankis , De Leon Daisy 
Published: October 2024

Abstract
Disclosure: K. Granados: None. A. Durán: None. F. Almaguel: None. D. De Leon: None.
Our laboratory previously showed that Insulin-like growth factor II (IGF-II) promotes cell proliferation, inhibits apoptosis and induces chemoresistance of Breast Cancer (BC) cells. We demonstrated that IGF-II achieves this by inhibiting pro-apoptotic proteins and stimulating anti-apoptotic proteins. Those studies also demonstrated that IGF-II prevented mitochondrial induced apoptosis by inhibiting depolarization of the mitochondrial membrane. Thus, IGF-II regulates the mitochondria to prevent cell death and to promote tumor growth and chemoresistance. Since IGF-II regulates mitochondrial proteins, we decided to assess how IGF-II regulates MAGMAS, a protein located in the inner mitochondrial membrane that is a critical component of a complex that controls mitochondrial protein import and ROS. Of note, MAGMAS was recently identified as being highly expressed in breast cancer, prostate cancer and during early fetal development which parallels IGF-II expression. Immunostaining of TNBC showed high levels of IGF-II and MAGMAS. Thus, our hypothesis is that IGF-II regulates MAGMAS and IGF-II secreting breast cancer cells will express higher levels of MAGMAS. Our preliminary results of rtPCR and Western blotting experiments validated our hypothesis demonstrating that IGF-II regulated the transcription of MAGMAS. To characterize the mechanisms by which IGF-II regulates MAGMAS, we chose the triple negative breast cancer (TNBC) cell line CRL-2335 which we demonstrated produces high levels of IGF-II and MAGMAS. CRL2335 cells were established from an AA TNBC patient and they express high levels of IGF-II and MAGMAS. Confocal microscopy and Western blotting and cellular fractionation were used to assess IGF-II regulation of MAGMAS in TNBC cells. The expression of MAGMAS was analyzed in wild CRL-2335, and IGF-II antisense transfected CRL-2335 cells and compared to cells treated with IGF-II. Results showed that CRL-2335 express high levels of MAGMAS in the mitochondria and blocking the expression of IGF-II by antisense technology significantly decreased MAGMAS. Exogenous IGF-II treatment to the antisense cells increased MAGMAS as demonstrated by Immunofluorescence and WB. Since IGF-II increases the localization of MAGMAS to the mitochondria we propose that IGF-II regulates MAGMAS to protect cancer cells from ROS induced cell death and chemoresistance. Completion of this study will validate IGF-II as a regulator of MAGMAS. Validation of IGF-II regulation of MAGMAS represents an important new tool for the treatment of TNBC.
Presentation: 6/2/2024
6603 Impact Of The Monocarbonyl Analogs Of Curcumin, C66 And B2BrBC, On The Protein Expression Of Epithelial And Mesenchymal Markers In MCF-7 Breast Cancer Cells
Authors: Stojchevski Radoslav , Lavi Aaron , Bogdanov Jane , Hadzi-Petrushev Nikola , Mladenov Mitko , Poretsky Leonid , Bogdanov Avtanski Dimiter 
Published: October 2024

Abstract
Disclosure: R. Stojchevski: None. A. Lavi: None. J. Bogdanov: None. N. Hadzi-Petrushev: None. M. Mladenov: None. L. Poretsky: None. D.B. Avtanski: None.
The synthesis of monocarbonyl analogs of curcumin (MACs) presents a solution to the challenges posed by curcumin, a natural polyphenol extracted from the roots of the turmeric plant (Curcuma longa), which is known for its antioxidant and anti-inflammatory properties. The limitations of curcumin, including poor stability, low bioavailability, and rapid metabolism, hinder its application, and MACs, designed without a β-diketone moiety, address these drawbacks and exhibit enhanced stability and bioavailability. This study aimed to assess the effects of two monocarbonyl analogs of curcumin, (2E,6E)-2,6-bis[(2-trifluoromethyl)benzylidene]cyclohexanone (C66) and (2E,6E)-2,6-bis(2-bromobenzylidene)cyclohexanone (B2BrBC), on epithelial-to-mesenchymal transition (EMT) in MCF-7 breast cancer cells using western blot analysis. Our previous findings indicated that these analogs can potentially affect EMT by modulating the epithelial and mesenchymal gene expression. MCF-7 cells were cultured in complete medium supplemented with EMT-inducing factors for two days, followed by C66 and B2BrBC administration (100 µM) for three additional days. After the experiments, proteins were extracted, and western blot analyses were performed to measure the expression levels of various epithelial and mesenchymal markers (E-cadherin, N-cadherin, SNAI1, and Claudin-1). The results revealed that both C66 and B2BrBC increased the protein levels of the epithelial marker E-cadherin and suppressed those of the mesenchymal marker N-cadherin. Moreover, C66 and B2BrBC decreased the protein expression of the transcription factor SNAI1 and transmembrane protein Claudin-1. Between the two MACs, B2BrBC showed a more pronounced effect on N-cadherin expression, while C66 was more potent in suppressing SNAI1 protein expression. Our findings demonstrate that C66 and B2BrBC counteract EMT, contributing to the potential therapeutic applications of MACs in managing breast cancer progression.
Presentation: 6/3/2024
7076 Inhibiting Oxidosqualene Cyclase, An Enzyme In The Cholesterol Biosynthetic Pathway, Suppresses Lung Metastasis Of Triple-Negative Breast Cancer Cells
Authors: Liang Yayun , Hyder Salman M 
Published: October 2024

Abstract
Disclosure: Y. Liang: None. S.M. Hyder: None.
Triple-negative breast cancers (TNBC) lack ER, PR and Her2neu proteins that are commonly targeted in breast cancer therapies. As a consequence, women who suffer from TNBC have poor prognosis and limited treatment options, and are usually administered toxic, non-specific chemotherapeutic agents. Drug-resistance almost always occurs, leading to tumor metastasis, the main cause of patient death. New non-toxic therapeutic strategies to control metastasis of TNBC are urgently needed. Cholesterol is an essential structural and functional component of cell membranes that is necessary for the growth of both primary tumors and metastatic colonies that reside in distant organs. Therefore, inhibiting cholesterol production is an attractive therapeutic strategy. Treatment with statins, a class of cholesterol biosynthesis inhibitors that target HMGCoA-reductase, is associated with certain undesirable side effects; consequently, we targeted oxidosqualene cyclase (OSC), an enzyme that occurs downstream of HMGCoA-reductase in the cholesterol biosynthetic pathway, to disrupt tumor metastasis. Potent small molecule inhibitors of OSC have been identified. RO 48-8071 (4′-[6-(allylmethylamino)hexyloxy]-4-bromo-2′-fluorobenzophenone fumarate) (RO), has emerged as a useful chemotherapeutic agent for treating several forms of primary tumors. We developed a mouse model for studying lung metastasis using a MDA-MB-231 variant (LM2) TNBC cell line obtained from Dr. Massagué (Memorial Sloan-Kettering Cancer Center). These are aggressive cells, and injection of only 2 X 105 cells produces detectable lung metastatic colonies in approximately 4-6 weeks. We inoculated female nu/nu mice, 5-6 weeks old, with 2 X 105 MDA-MB-231/LM2 cells in 0.1 ml DMEM/F12 medium without serum via tail vein. Five days after tumor cell inoculation animals received 20 mg/kg (n=6), 40 mg/kg (n=7) RO (ip) or vehicle alone (n=7) every other day for two weeks. Animals were then sacrificed, and both lungs harvested and immediately fixed in Bouin’s fixative. Metastatic colonies on the lung surface were identified and counted under a dissecting microscope. Our study showed that RO effectively reduced lung metastasis without affecting animal weight. Compared with 56 + 7 colonies (Mean ± SEM) in controls given vehicle alone, RO significantly reduced lung metastatic colonies to 28 ± 5 in animals exposed to 20 mg/kg and 6 ± 1 in 40 mg/kg treated animals (p&lt;0.05, ANOVA). Reduction in number of colonies in 40 mg/kg group was also significantly different from colonies counted in 20 mg/kg treated animals. Our study is the first to demonstrate that disruption of cholesterol biosynthesis in a model of human TNBC leads to reduced metastasis in lungs. Further studies will determine mechanisms through which RO suppresses TNBC metastasis to the lungs. Supported by Zalk Missouri Professor Endowment Funds.
Presentation: 6/3/2024