Proteomic profiling reveals novel insights into hypercortisolism adrenal adenomas.

A Pilot Study on Serum Lipidomic Alterations in Patients with Adrenal Tumors

AD18 Integrated steroidomic and untargeted metabolomic profiling reveals distinct circulating signatures of adrenocortical carcinoma

Adrenal tumours display marked biological heterogeneity, making the distinction between benign and malignant lesions challenging. Liquid chromatography–high-resolution mass spectrometry (LC-HRMS) enables simultaneous investigation of steroidomic and broader metabolomic alterations. To identify integrated circulating steroidomic and metabolomic signatures associated with adrenocortical carcinoma (ACC), lipid-poor adenomas (LPAs), and lipid-rich adenomas (LRAs). Basal serum samples were collected from patients with ACC (n = 33), LPA (n = 68), and LRA (n = 166). Following protein precipitation and solid-phase extraction, 100 μL of serum was analysed using an in-house LC-HRMS method on the Orbitrap Exploris 240 platform (Thermo Fisher Scientific). Targeted absolute quantification was performed for 45 adrenal steroids, while full-scan acquisitions were processed using Compound Discoverer software for untargeted metabolite identification. Thirty-one steroids were detected, of which 14 differed significantly among the study groups. Untargeted analysis identified 806 features with annotation confidence >80%, including 34 endogenous metabolites with significant discriminatory value. Compared with LRA and LPA, ACC was characterized by significantly higher concentrations of pregnenolone, 17-hydroxypregnenolone, progesterone, 17-hydroxyprogesterone, 16-hydroxyprogesterone, 11β-hydroxyprogesterone, 11-deoxycorticosterone, 11-deoxycortisol, androstenedione, allopregnanolone, and dehydroepiandrosterone sulfate (DHEAS) (P < 0.001–0.034). ACC was also associated with increased arachidonic acid, docosahexaenoic acid, and several oxylipins, including hydroxyeicosatetraenoic acids, leukotriene derivatives, and resolvin D4 (all P < 0.001). In contrast, LPAs demonstrated higher aldosterone and 20β-dihydrocortisone concentrations, together with lower α-linolenoyl ethanolamide and glycerolipid levels than LRAs (all P < 0.007). Adrenocortical carcinoma exhibits a distinct circulating molecular signature characterized by dysregulated steroidogenesis together with enhanced lipid oxidation and inflammatory signalling. Although LPAs and LRAs showed largely overlapping metabolic profiles, several discriminatory biomarkers were identified. Integration of steroidomic and metabolomic profiling may improve the characterization of adrenal lesions and facilitate the identification of ACC.

Liver cirrhosis is associated with altered glucocorticoid metabolism related to disease severity.

BACKGROUND Liver cirrhosis (LC) is frequently complicated by adrenal dysfunction; however, whether cirrhosis drives the coordinated, pathway-specific reprogramming of cortisol metabolism remains unclear. OBJECTIVES We aimed to characterize axis-specific alterations in cortisol metabolism in patients with LC, those with adrenal insufficiency (AI), and controls using integrated serum and salivary steroid profiling. METHODS This prospective study included adults with LC (n = 33) and control participants without cirrhosis who underwent cosyntropin-stimulation test following transsphenoidal surgery for a nonfunctioning pituitary adenoma (controls, n = 27; AI, n = 10). Serum and saliva samples were collected at baseline, 30 and 60 min after stimulation and profiled for 26 serum and 12 salivary steroids through liquid chromatography-tandem mass spectrometry. Multivariate models were constructed to identify cirrhosis-associated steroid features, and their associations with disease severity. RESULTS LC was characterized by axis-specific steroid remodeling, including suppressed 11β-hydroxysteroid dehydrogenase 1 and A-ring reduction activity with the 20α-reduction pathway activation. Representative changes included a lower peak cortisol-to-cortisone ratio and reduced tetrahydrocortisone-to-cortisone ratio, alongside increased 20α-reduced metabolites (all p < 0.001). This pattern was distinct from that in AI, in which steroid production and downstream metabolites were uniformly reduced. Iterative model refinement identified a parsimonious seven-steroid signature that discriminated patients with LC from controls. Within the LC cohort, the steroid signature correlated with the Child-Pugh score. CONCLUSION LC is associated with coordinated, axis-specific cortisol metabolic reprogramming, which is mechanistically distinct from AI. Integrated steroid profiling may provide mechanistic insight into altered glucocorticoid metabolism in cirrhosis and its relationship to disease severity .

Adrenal incidentalomas: 1 mg dexamethasone test result in the context of cardiometabolic risk.

PURPOSE Mild autonomous cortisol secretion (MACS) is the most common hormonal abnormality in adrenal incidentalomas. The 1 mg dexamethasone suppression test (DST) with a cutoff of 1.8 µg/dL is used to exclude hypercortisolism, yet some "non-functioning" tumors may secrete cortisol at concentrations below this threshold, potentially contributing to an adverse cardiometabolic profile. The aim of this study was to evaluate this relationship. METHODS In this retrospective cross-sectional study, data from 1,077 patients with adrenal incidentalomas treated at a tertiary endocrinology center between 2017 and 2020 were analyzed. After applying the inclusion criteria, 807 patients were included. All patients underwent a 1 mg DST. Receiver operating characteristic (ROC) curves and area under the curve (AUC) analyses were used to assess associations between post-DST cortisol concentrations and cardiometabolic conditions, and to identify a threshold for a composite endpoint comprising the presence of coronary heart disease, chronic heart failure, or diabetes mellitus. RESULTS Patients with hypertension, coronary heart disease, heart failure, diabetes mellitus, chronic obstructive pulmonary disease, and chronic kidney disease had significantly higher post-DST cortisol concentrations. However, in multivariable logistic regression, post-DST cortisol was not independently associated with the composite endpoint after adjustment for confounders, with age and obesity remaining significant predictors. Exploratory analysis suggested that cortisol concentrations > 1.37 µg/dL were associated with a higher prevalence of the composite endpoint. CONCLUSION Post-DST cortisol concentrations below the standard cutoff may be associated with cardiometabolic comorbidities in patients with adrenal incidentalomas. These findings should be interpreted with caution and require confirmation in prospective studies.

NE07 A simple clinical and biochemical prediction score for distinguishing ectopic ACTH syndrome from Cushing disease

Differentiating ectopic ACTH syndrome (EAS) from Cushing disease (CD) remains challenging in ACTH-dependent Cushing syndrome. We aimed to evaluate routinely available markers and develop a simple prediction score for distinguishing EAS from CD. This retrospective single-center study included 85 patients with ACTH-dependent Cushing syndrome: 77 with CD and 8 with EAS. Clinical, biochemical, and imaging findings were compared. Receiver operating characteristic analyses were used to assess marker performance, and the most discriminative variables were incorporated into the Ectopic Cushing Prediction Score (ECPS). Patients with EAS had a more severe phenotype, with higher rates of hypertension, proximal muscle weakness, hypokalemia, and opportunistic infections. ACTH, serum cortisol, midnight cortisol, late-night salivary cortisol (LNSC), and 24-hour urinary free cortisol (UFC) were significantly higher in EAS than in CD. Among individual markers, hypokalemia showed the highest diagnostic accuracy (AUC=0.967; sensitivity 100%; specificity 93.3%), followed by UFC >10× ULN (AUC=0.950) and ACTH (AUC=0.891). Optimal cut-offs were 467.5 μg/24 h for UFC and 149.5 pg/mL for ACTH. The ECPS included hypokalemia, UFC >10× ULN, ACTH ≥149.5 pg/mL, LNSC ≥1.4× ULN, and post–1 mg dexamethasone suppression test cortisol ≥14.85 μg/dL. The ECPS showed excellent performance (AUC=0.994), and a score ≥5 yielded 87.5% sensitivity and 98.6% specificity. The ECPS accurately differentiates EAS from CD using routine clinical and biochemical parameters and may guide the diagnostic evaluation of ACTH-dependent Cushing syndrome.

Discovery of novel protein biomarkers for the detection of polyendocrine metabolic ovarian syndrome phenotype A

Polyendocrine metabolic ovarian syndrome (PMOS) is a prevalent endocrine condition affecting women of reproductive age. PMOS is underrecognized/underdiagnosed and associated with secondary comorbidities that may result from delayed diagnosis/management. Novel biomarkers are needed to improve timely diagnosis. This retrospective, case-control study used biobanked serum samples from women with PMOS phenotype A ( n  = 51) and healthy controls ( n  = 37). We performed protein biomarker discovery using Olink Proximity Extension Assay and receiver operating characteristic-area under the curve (ROC-AUC) analysis. We conducted an overrepresentation pathway analysis against Reactome. We investigated 1,196 protein biomarkers and conducted a pathway analysis using the 145 top-performing biomarkers (AUC > 0.750). Top 10 performing biomarkers (AUC ≥ 0.961) were linked to relevant biological pathways, including immune system regulation (serum amyloid A4 and leukotriene A4 hydrolase), lipid, carbohydrate, and protein metabolism (fibroblast growth factor binding protein 1, carboxylesterase 2, inositol polyphosphate-1-phosphatase, inositol polyphosphate-1-phosphatase like, pro-glucagon, and leptin), nervous system development (semaphorin 4C and plexin B1), and cellular response (peroxiredoxin 1). These pathways/sub-pathways may contribute to PMOS pathophysiology and related comorbidities (insulin resistance, obesity, and fatty liver disease). We discovered novel circulating protein biomarkers that distinguish women with PMOS phenotype A from healthy controls, supporting diagnosis and enabling timely intervention/appropriate management.