Specific characteristics of metabolomics as assessed by gas chromatography-mass spectrometry in patients with adrenocortical cancer and with adrenal incidentalomas in congenital adrenal hyperplasia

Background: Prolonged episodes of uncontrolled congenital adrenal hyperplasia (CAH) have been shown to result in the occurrence of secondary adrenal neoplasms. Prevalence of adrenal incidentalomas in the patients with 21-hydroxylase deficiency ranges from 11% to 82%. As assessed by gas chromatography-mass spectrometry (GC-MS), patients with adrenocortical cancer (ACC) have increased level of steroid hormone precursors due to decreased activity of adrenal steroidogenesis enzymes, mainly that of 21-hydroxylase and 11-hydroxylase. It seems relevant to compare the specific characteristics of steroid metabolism by GC-MS in ACC patients and in patients with adrenal incidentalomas and CAH associated with 21-hydroxylase deficiency (21-OHD). Aim: To identify (by GC-MS) common abnormalities in steroid metabolism and differential diagnostic biomarkers in ACC patients and CAH patients with 21-OHD and adrenal masses. Materials and methods: The study included 41 patients with adrenal cortex neoplasms aged 18 to 65 years without clinical and laboratory signs of endogenous hypercortisolism. Twenty three (23) patients had non-metastatic ACC and 18 patients had CAH due to 21-OHD. The control group included 26 healthy blood donors aged 20 to 59 years. Urine steroid profiles were measured by GC-MS with a gas chromatograph-mass spectrometer (Shimadzu GCMS-QP2020). Results: In the ACC patients, there was an increase in urinary excretion of tetrahydro-11-deoxycortisol, dehydroepiandrosterone, androstenediol-17, etiocholanolone, pregnenediol, and 3,16,20-pregnenetriol (3,16,20-dP3), as well as a decrease in the 3,16,20-dP3/3,16,20-dP3 ratio, compared to the values in the patients with CAH due to 21-OHD. Compared to the healthy control, 21-hydroxylase, 11-hydroxylase, 5-reductase and 11-hydroxysteroid-dehydrogenase (11-HSDH) type 2 activities were lower. Compared to the ACC patients, those with CAH due to 21-OHD had higher urinary excretion of 11-oxo-pregnanetriol (11-oxo-P3) and 21-deoxy-tetrahydrocortisol and lower 5-THF+5-THF+THE)/11-oxo-P3 ratio of 9.0, determination of 11-oxo-dP3, signs higher 5-reductase activity and lower 11-HSDH type 1 activity. The ACC patients and the patients with CAH due to 21-OHD had common abnormalities of steroid metabolism, such as lower activities of 21-hydroxylase, 3-hydroxysteroid-dehydrogenase and 11-hydroxylase, and no differences in urinary excretion of a number of ACC biomarkers (androgens, pregnanediol, and 5-ene-pregnenes). Conclusion: The assessment of urinary excretion of androgens, progestagens, and glucocorticoids by GC-MS made it possible to identify common abnormalities in steroid metabolism in the patients with ACC and CAH due to 21-OHD, which confirms the role of disordered steroidogenesis in the formation of adrenocortical tumors.