Validation of prognostic biomarkers in adrenocortical carcinoma through a Next-Generation Sequencing in real-life setting

Abubaker Mohamed; Lorenzo Tucci; Biase Dario De; Juliane Lippert; Lisa James; Nicola Chadderton; Alice Fair; Kassiani Skordilis; Leo Antonio De; Phillipe Taniere; Dalmazi Guido Di; Cristina L Ronchi

doi:10.1530/endoabs.110.P10

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Endocrine Abstracts (2025) 110 P10 | DOI: 10.1530/endoabs.110.P10

ECEESPE2025 Poster Presentations Endocrine Related Cancer (76 abstracts)

Validation of prognostic biomarkers in adrenocortical carcinoma through a Next-Generation Sequencing in real-life setting

Abubaker Mohamed ¹ , Lorenzo Tucci ^2,3,4 , Dario De Biase ^5,6 , Juliane Lippert ⁷ , Lisa James ¹ , Nicola Chadderton ¹ , Alice Fair ¹ , Kassiani Skordilis ⁸ , Antonio De Leo ^4,6 , Phillipe Taniere ¹ , Guido Di Dalmazi ^3,4 & Cristina L Ronchi ^2,9

Author affiliations

¹Queen Elizabeth Hospital Birmingham, University Hospitals Birmingham NHS Foundation Trust, Molecular Pathology Diagnostic Service, Birmingham, UK; ²University of Birmingham, Department of Metabolism and Systems Science, Birmingham, UK; ³IRCCS Sant’Orsola Polyclinic, Division of Endocrinology and Diabetes Prevention and Care, Bologna, Italy; ⁴Alma Mater Studiorum University of Bologna, Department of Medical and Surgical Sciences (DIMEC), Bologna, Italy; ⁵University of Bologna, Department of Pharmacy and Biotechnology, Bologna, Italy; ⁶IRCCS Azienda Ospedaliero-Universitaria di Bologna, Solid Tumor Molecular Pathology Laboratory, Bologna, Italy; ⁷University of Wuerzburg, Institute of Human Genetics, Wuerzburg, Germany; ⁸Queen Elizabeth Hospital Birmingham, University Hospitals Birmingham NHS Trust, Department of Histopathology, Birmingham, UK; ⁹Queen Elizabeth Hospital Birmingham NHS Trust, Department of Endocrinology, Birmingham, UK

JOINT1372

Background: Adrenocortical carcinoma (ACC) is a rare aggressive malignancy with heterogenous clinical outcomes. Our previous research demonstrated that targeted DNA-based biomarkers assessed on paraffin-embedded (FFPE) samples can improve prognostic classification.

Aim: To investigate the feasibility in real-life setting of a Next-Generation Sequencing (NGS) service for prognostic classification of ACC in two European centres (Queen Elizabeth Hospital Birmingham (QEHB), UK, and IRCCS Sant’Orsola Polyclinic, Bologna, Italy).

Methods: A total of 43 patients with ACC were enrolled, who underwent adrenalectomy at QEHB (n=23) or IRCCS Sant’Orsola Polyclinic (n=20), including a previously published retrospective cohort (n=10; surgery date 2012–2020) and an independent validation cohort (n=33; surgery date 2002–2024). Tumour DNA was extracted from FFPE tissue using a Qiagen DNA protocol and quantified by Qubit fluorometer. A targeted NGS platform for 10 ACC-specific genes (ATM, APC, RB1, MEN1, CDK4, NF1, ZNRF3, TERT promoter, TP53 and CTNNB1) was developed and validated using the retrospective cohort and commercially available DNA control (HD827, Horizon Oncospan). Sequencing was performed using an Illumina platform. The variant allele frequency (VAF) threshold was validated at 5% at QEHB and 10% at IRCCS Sant’Orsola Polyclinic. We assessed predicted turnaround time (TAT) and costing of implementing this NGS service within a routine laboratory setting.

Results: After 11 cases were excluded due to low quality DNA, the final cohort consisted in 32 patients (25F/7M, average age 50yrs). In the retrospective cohort (n=6), all previously identified somatic variants (i.e. in RB1, MEN1, ZNRF3, CTNNB1 and NF1) were confirmed. Two more variants were detected in the added TERT promotor region c.−146.C>T and c.−124C>T. In the validation cohort, 36 variants were reported in 14/26 patients (53.8%). Pathogenic/likely pathogenic variants (52.8% of total variants) were detected in TP53 (22.2%, mean VAF 50.6%), NF1 (19.4%, mean VAF 41.73%), CTNNB1 (p.Ser45Pro, mean VAF 62.3%, 11.1%), with variants of uncertain significance found in TERT, APC, MEN1, NF1 and ZNRF3 (13.9%). The predicted TAT for the validation cohort was 21 calendar days from sample receipt to final report, with an average 1.5 hours reporting time per case. There was no relevant difference for full costings between the two centres (C= 200–250 per sample).

Conclusions: Our proposed ACC-specific NGS service was reliable, feasible and cost-effective for most cases allowing implementation in real-life clinical setting. Most detected variants have shown strong evidence for clinical significance, prognostic value and/or represent potential drug targets (NF1), potentially facilitating personalised management of patients with ACC.