ccfDNA analysis for the classification of adrenocortical adenomas

• Published in: Journal of endocrinological investigation Vol. 48; no. 5; pp. 1207 - 1216
• Main Authors: Xu, Mengjie, Tourigny, David S., Lippert, Juliane, Crastin, Ana, Appenzeller, Silke, Asia, Miriam, Podstawka, Oskar, Smith, Gabrielle, Elhassan, Yasir S., Skordilis, Kassiani, Prete, Alessandro, Ronchi, Cristina L.
• Format: Journal Article
• Language: English
• Published: Cham Springer International Publishing 01.05.2025; Springer Nature B.V
• Subjects: Adenoma; Adrenal Cortex Neoplasms - blood; Adrenal Cortex Neoplasms - classification; Adrenal Cortex Neoplasms - diagnosis; Adrenal Cortex Neoplasms - genetics; Adrenal Cortex Neoplasms - pathology; Adrenocortical Adenoma - blood; Adrenocortical Adenoma - classification; Adrenocortical Adenoma - diagnosis; Adrenocortical Adenoma - genetics; Adrenocortical Adenoma - pathology; Adult; Aged; Aldosterone; Biomarkers, Tumor - blood; Biomarkers, Tumor - genetics; Case-Control Studies; Cell-Free Nucleic Acids - blood; Cell-Free Nucleic Acids - genetics; Cortisol; Cushing syndrome; Cushing Syndrome - genetics; Endocrinology; Female; Follow-Up Studies; Hormones; Humans; Hydrocortisone - metabolism; Internal Medicine; Male; Medicine; Medicine & Public Health; Metabolic Diseases; Middle Aged; Mutation; NF-AT protein; Nucleotide sequence; Original; Original Article; Prognosis; T-DNA; Tumors; Circulating cell-free DNA; Liquid biopsy; Adrenocortical tumours; Next-generation sequencing; Primary aldosteronism; Cortisol excess
• ISSN: 1720-8386; 0391-4097; 1720-8386
• DOI: 10.1007/s40618-025-02540-5

Background Somatic alterations are commonly observed in adrenocortical adenomas including cortisol-producing (CPA) [overt Cushing syndrome (CS) or mild autonomous cortisol secretion (MACS)], aldosterone-producing (APA), and non-functioning (NFAT) tumors. We tested whether somatic variants could be detected in circulating cell-free DNA (ccfDNA) from patients with adenomas and potentially contribute to management strategies. Materials and methods We investigated 44 patients (17 CPA-MACS, 9 CPA-CS, 12 APA, and 6 NFAT). 23 healthy subjects (HS) served as controls. ccfDNA was extracted from blood samples and quantified with fluorimeter. Tumor DNA (T-DNA) was isolated from paraffin embedded tissue in 17/44 cases. Matched ccfDNA/T-DNA were sequenced using a customized panel including 32 genes. Leucocyte DNA was used to filter out germline variants. Results Patients with adenomas had higher total ccfDNA concentrations than HS [median 0.12 (IQR 0.05–0.19) vs. 0.05 (0.00-0.08) ng/µl, P  < 0.001], with CPA-CS showing the highest ccfDNA levels [0.18 (0.05–0.47) ng/µl]. Within T-DNA, somatic variants were identified in 53% of adenomas: PRKACA in 2/7 CPA-CS, CTNNB1 in 3/5 CPA-MACS and 1/7 CPA-CS, KCNJ5 in 2/5 APA and CACNA1D in 1/5 APA. Somatic mutations were not detected in any of the investigated ccfDNA samples. Conclusions Total ccfDNA concentrations are higher in patients with CPA-CS. Despite the presence of somatic variants in half of tumor samples, we did not detect any at ccfDNA level. Therefore, this approach appears ineffective for pre-operative detection of genetic alterations.