Baseline biochemical evaluation for etiologic differentiation in ACTH-dependent Cushing’s syndrome: machine learning and composite score analysis in a multicenter cohort of 566 patients.

BACKGROUND: Limited availability of corticotropin-releasing hormone (CRH) currently complicates the differentiation of adrenocorticotropin (ACTH)-dependent Cushing's syndrome (CS). The diagnostic value of common screening tests in distinguishing Cushing's disease (CD) from ectopic CS (ECS) remains unclear. OBJECTIVE: To assess the diagnostic performance of screening tests, alone and in combination, in differentiating CD from ECS. METHODS: Retrospective multicenter study enrolling patients with confirmed ACTH-dependent CS and available screening tests at diagnosis. Data are expressed as multiples of upper limit of normal (×ULN). Optimal cut-offs were determined using Youden's Index. Combination with composite score models and machine learning (ML) algorithm were performed. RESULTS: A total of 566 patients were included (509 [90%] with CD). The optimal morning ACTH cut-off was 1.8×ULN (sensitivity 74%, specificity 77%, AUC=0.776 [CI-95% 0.688-0.853]). 24h-urinary free cortisol (24h-UFC) showed the best performance (cut-off 5.9×ULN, sensitivity 72%, specificity 83%, AUC=0.854 [0.816-0.923]), followed by the 1 mg dexamethasone suppression test (12.8×ULN, sensitivity 70%, specificity 86%, AUC=0.828 [0.740-0.906]). Using a composite score, a cut-off of 1.5 yielded 78% sensitivity and 92% specificity (AUC=0.865 [0.798-0.933]). Combining this score with pituitary MRI-findings, sensitivity and specificity were 88% and 85% (AUC=0.931 [0.887-0.974]). ML algorithm (balanced random forest) including screening tests-results yielded a sensitivity of 71% and specificity of 84% (AUC=0.853 [0.741-0.966]), while including the MRI-findings variant achieved a sensitivity and specificity of 76% and 91% (AUC=0.902 [0.805-0.999]). CONCLUSION: Combining screening tests in composite scoring and ML differentiates ACTH-dependent CS and offers a potential diagnostic alternative to CRH-stimulation test.