MicroRNA Expression Profiling in Adrenal Myelolipoma.

Introduction: Adrenal myelolipoma (AML) is the second most common, and invariably benign primary adrenal neoplasm. Due to the variable proportion of fat and hematopoietic elements, and its often large size, it can cause differential diagnostic problems. Several reports confirmed the utility of microRNAs (miRNAs) in the diagnosis of tumors, but the miRNA expression in AML has not yet been investigated. Materials and methods: Next-generation sequencing (NGS) was performed on 30 formalin-fixed paraffin-embedded (FFPE) archived tissue [AML, adrenocortical adenoma (ACA) and adrenocortical carcinoma (ACC) 10 each] samples. Validation was performed by real-time RT-qPCR on a cohort containing 41 further FFPE samples (15 AML, 14 ACA and 12 ACC). Circulating miRNA counterparts of significantly differentially expressed tissue miRNAs were studied in altogether 33 plasma samples (ACA, ACC, AML 11 each). Results: By NGS, 256 significantly differentially expressed miRNAs were discovered, and 8 of these were chosen for validation. Significant overexpression of hsa-miR-451a, hsa-miR-486-5p, hsa-miR-363-3p and hsa-miR-150-5p was confirmed in AML relative to ACA and ACC. Hsa-miR-184, hsa-miR-483-5p and hsa-miR-183-5p were significantly overexpressed in ACC relative to ACA, but not to AML. Circulating hsa-miR-451a and hsa-miR-363-3p were significantly overexpressed in AML, whereas circulating hsa-miR-483-5p and hsa-miR-483-3p were only significantly overexpressed in ACC vs. ACA. Conclusions: We have found significantly differentially expressed miRNAs in AML and adrenocortical tumors. Circulating hsa-miR-451a might be a promising minimally invasive biomarker of AML. The lack of significantly different expression of hsa-miR-483-3p and hsa-miR-483-5p between AML and ACC might limit their applicability as diagnostic miRNA markers for ACC

A növényi szteroid hormon bioszintézis szabályozásának molekuláris genetikai vizsgálata = Molecular genetic dissection of the regulation of plant steroid hormone biosynthesis

A támogatási időszak során a brasszinoszteroidok (BR-ok, növényi szteroid hormonok) bioszintézisében résztvevő gének funkcióit és szabályozását vizsgáltuk Arabidopsis modellnövényben. Molekuláris genetikai módszerek segítségével meghatároztuk a CYP90C1 és CYP90D1 gének által kódolt ezimek szerepét a hormon szintézisében, és eredményeink alapján új bioszintézis modellt dolgoztunk ki. Elvégeztük a BR szintézis kulcsenzimeit kódoló CPD/CYP90A1 és CYP85A2 gének kifejeződésének részletes vizsgálatát, és kimutattuk, hogy expressziós szintjük emelkedése az aktív hormon felhalmozódását eredményezi. Mindez arra utal, hogy a lokális BR szint kialakításában a bioszintézis génjinek transzkripciós szintű szabályozása fontos szerepet játszik. E két gén napszakos szabályozásának tanulmányozása során bizonyítottuk a fény induktív szerepét, és tisztáztuk hogy a fényhatás közvetítésében a fitokróm B fotoreceptorról kiinduló szignálátviteli útnak van meghatározó jelentősége. | In the frame of the current project we studied the functions and regulation of the genes that are involved in the biosynthesis of brassinosteroids (BRs, plant steroid hormones) in Arabidopsis. Using molecular genetic approaches, we identified the reactions catalyzed by the CYP90C1 and CYP90D1 genes, and based on these results we revised the existing model of the BR biosynthetic pathway. We carried out a detailed expressional analysis of the CPD/CYP90A1 and CYP85A2 genes that encode rate limiting enzymes of BR synthesis, and demonstrated that their increased activity results in the accumulation of the bioactive hormone. These results strongly suggest a key role for the transcriptional regulation of BR-biosynthetic genes in the setting of local BR levels. Studies of the diurnal regulation of these genes revealed the inductive role of light, and we determined that this light control depends primarily on signals originating from the phytochrome B photoreceptor. Our results strongly suggest a key role for the transcriptional regulation of BR-biosynthetic genes in the setting of local BR levels

Flg22-Triggered Immunity Negatively Regulates Key BR Biosynthetic Genes

In plants, activation of growth and activation of immunity are opposing processes that define a trade-off. In the past few years, the growth-promoting hormones brassinosteroids (BR) have emerged as negative regulators of pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI), promoting growth at the expense of defense. The crosstalk between BR and PTI signaling was described as negative and unidirectional, since activation of PTI does not affect several analyzed steps in the BR signaling pathway. In this work, we describe that activation of PTI by the bacterial PAMP flg22 results in the reduced expression of BR biosynthetic genes. This effect does not require BR perception or signaling, and occurs within 15 min of flg22 treatment. Since the described PTI-induced repression of gene expression may result in a reduction in BR biosynthesis, the crosstalk between PTI and BR could actually be negative and bidirectional, a possibility that should be taken into account when considering the interaction between these two pathways

The Clinical Impact of [68Ga]‐DOTATATE PET/CT for the Diagnosis and Management of Ectopic Adrenocorticotropic Hormone – Secreting Tumours

ObjectivesLocalization of ectopic ACTH‐secreting tumours causing Cushing syndrome (ECS) is essential for clinical management, yet often difficult. [68Ga]‐DOTATATE PET/CT ([68Ga]‐DOTA‐(Tyr3)‐octreotate)] is an FDA‐approved high‐resolution diagnostic tool for imaging neuroendocrine tumours. Data on the clinical utility of [68Ga]‐DOTATATE in patients with ECS, however, are scarce. The objectives of this study were to determine the efficacy for ECS localization and the clinical benefit of [68Ga]‐DOTATATE imaging.MethodWe conducted a retrospective review of all cases with ECS evaluated with [68Ga]‐DOTATATE from November 2016 through October 2018 at three referral centres. The clinical benefit of [68Ga]‐DOTATATE was based on detection of new tumours and resultant changes in management.ResultsOver the study period, 28 patients with ECS underwent [68Ga]‐DOTATATE: 17 for identification of the primary tumour and 11 during follow‐up. [68Ga]‐DOTATATE identified the suspected primary ECS in 11/17 patients (65%). Of these, nine patients underwent surgery: eight with confirmed ECS (5 bronchial, 1 thymic, 1 pancreatic and 1 metastatic neuroendocrine tumour of unknown primary origin) and one patient with a false‐positive scan (adrenal gland). Of the 11 patients with ECS who underwent [68Ga]‐DOTATATE evaluation during follow‐up, the study led to changes in clinical management in 7/11 (64%) patients.Conclusions[68Ga]‐DOTATATE is sensitive in detecting primary and metastatic ECS, often identifies occult tumours after conventional imaging, and impacts clinical care in the majority of patients.Peer Reviewedhttps://deepblue.lib.umich.edu/bitstream/2027.42/150507/1/cen14008.pdfhttps://deepblue.lib.umich.edu/bitstream/2027.42/150507/2/cen14008_am.pd

Incidence of Venous Thromboembolic Events in Patients With Endogenous Cushing Syndrome

Background: Hypercortisolemia is a hypercoagulable state associated with increased risk of venous thromboembolic events (VTE). The reported incidence of VTE in patients with ACTH-dependent or independent Cushing Syndrome (CS) is variable, ranging from 3 to 14%. Our aim was to assess the incidence of clinically significant VTE among patients with endogenous CS and to identify risk factors for the development of VTE. Methods: We conducted a single center retrospective longitudinal study of adult patients diagnosed with endogenous CS between 2010 and 2020. Patients with a known prothrombotic disease (e.g. Factor V Leiden), insufficient data, or non-neoplastic hypercortisolism were excluded. Data collected included patient demographics, presenting symptoms, biochemical and radiological workup, treatment details, and incidence of clinically significant VTE. Results: A total of 114 patients (mean age of 45.55 ± 14.78 years, 79.8% women) followed for mean of 3.26 ± 2.9 years were included. Of the 114 patients, 58 (50.9%) had Cushing disease (CD), 40 (35.1%) had CS due to adrenal adenoma/hyperplasia, 6 (3.5%) had adrenocortical carcinoma (ACC), and 10 (8.8%) had ectopic Cushing syndrome (eCS). The overall incidence of VTE at any time point was 14/114 (12.3%); 11 (79%) VTEs were associated with presence of an additional VTE risk factor (8 surgery and 3 malignancy). Prior to any intervention for CS, 3 of 114 (2.6%) patients had a VTE. Surgery for CS (adrenalectomy, transsphenoidal surgery, tumor resection) was performed in 97 patients (85.1%) whereas 17 were treated medically (n=10), died before treatment (n=1) or observed (n=6). VTE occurred in 2 patients receiving medical therapy for CS. The post-operative incidence of VTE was 9 (9.3%; 4 in CD, 1 in adrenal CS, 3 in ACC, and 1 in eCS). VTE occurred ≤ 3-month post-operative in 4 patients (44.4%). Among the 5 patients in whom VTE occurred >3 months post-operative, 3 had recurrent metastatic ACC with hypercortisolemia and 2 were in remission (1 with CS and 1 with eCS). The median time from surgery to VTE occurrence was 315 days (8-1006). Compared to those who did not develop VTE, those who developed VTE had higher mean 24-hour urine free cortisol (4663.6 vs 558.21 mcg/dL; n = 100, P < 0.0001) and mean 1 mg overnight dexamethasone suppression test (36.3 vs 11.8 mcg/dL; n = 69, P = 0.0003), but similar mean late-night salivary cortisol (0.591 vs 0.790 ng/dL, n = 84, P = 0.71) at diagnosis of CS. Discussion: Among those with CS, the overall incidence of VTE was 12.3% and the majority of VTE were provoked (surgery, malignancy). Moreover, VTE was more likely in those with higher UFC and 1 mg overnight dexamethasone suppression test in our cohort. This suggests that in patients with CS who have an active malignancy, severe CS or those undergoing a surgical procedure may be at increased risk of VTE. Future studies should investigate the optimal type and duration of the VTE prophylaxis

Analysis of circulating extracellular vesicle-associated microRNAs in cortisol-producing adrenocortical tumors

PURPOSE: Circulating microRNAs (miRNA) have been described in patients with adrenocortical tumors, but the expression of miRNAs in non-functioning and cortisol-producing tumors has not been yet compared. Therefore, the objective of this study was to evaluate the expression of plasma extracellular vesicle (EV)-associated microRNAs in patients with non-functioning adrenocortical adenoma (NFA), cortisol-producing adrenocortical adenoma (CPA) and cortisol-producing adrenocortical carcinoma (CP-ACC). METHODS: Preoperative plasma EV samples of 13 NFAs, 13 CPAs and 9 CP-ACCs were subjected to extracellular vesicle isolation. miRNAs were investigated by targeted quantitative real-time PCR normalized to cel-miR-39 as reference. Five miRNAs have been selected for this analysis based on the previous studies including hsa-miR-22-3p, hsa-miR-27a-3p, hsa-miR-210-3p, hsa-miR-320b and hsa-miR-375. RESULTS: We have observed significant overrepresentation of three miRNAs in both CPA and CP-ACC relative to NFA: hsa-miR-22-3p (p < 0.01 and p < 0.0001, respectively), hsa-miR-27a-3p (p < 0.05 in both comparisons) and hsa-miR-320b (p < 0.05 and p < 0.0001, respectively). Hsa-miR-320b has been significantly overrepresented in CP-ACC relative to CPA (p < 0.01). Hsa-miR-210-3p turned out to be significantly overrepresented only in CP-ACC compared to NFA (p < 0.05). Significant correlation was revealed between circulating miRNA concentrations and urinary free cortisol values for hsa-miR-22-3p, hsa-miR-27a-3p and hsa-miR-320b (p < 0.0001 for all) and cortisol after low-dose dexamethasone test for hsa-miR-22-3p and hsa-miR-320b (p < 0.05). Hsa-miR-27a-3p has been significantly stimulated by low-dose dexamethasone test (p < 0.05). CONCLUSIONS: EV-associated miRNAs are differentially expressed in different non-functioning and cortisol-producing adrenocortical tumors