Circulating large extracellular vesicles as diagnostic biomarkers of indeterminate thyroid nodules - multi-platform omics analysis

Q: Why are indeterminate thyroid nodules (Thy3f/Bethesda IV) clinically challenging?

Most nodules are benign, but 7–15% are malignant, and Thy3f carries an estimated malignancy risk of 15–30%. Current diagnostic hemithyroidectomy is used to confirm histology, creating need for better non-invasive tests.

Q: How were circulating large extracellular vesicles measured in this study?

Plasma large extracellular vesicle concentrations were quantified using flow cytometry. miRNA profiles were identified with next-generation sequencing, and protein profiles were assessed using mass spectrometry.

Q: What key biomarker differences were found between Thy3f nodules and healthy controls?

Total large extracellular vesicle concentrations were higher in Thy3f patients. Vesicles expressing epithelial cell adhesion molecule and several cancer-related markers showed significantly increased concentrations versus healthy individuals.

Q: What was the study’s main conclusion for diagnostic use?

Circulating large extracellular vesicle miRNA and protein profiles demonstrated high diagnostic value to distinguish benign from malignant Thy3f nodules, with further validation required for clinical performance.

Sep 21, 2026

Uploaded by Theodora

Background: Most thyroid nodules are benign, yet 7–15% are malignant. Patients with indeterminate thyroid nodules (Bethesda IV/Thy3f) often undergo diagnostic hemithyroidectomy for definitive histology. Methods: This two-gate diagnostic accuracy study matched Thy3f patients (by age, sex, BMI) to healthy controls. Final histology established benign versus malignant outcomes. Plasma large extracellular vesicle counts were quantified by flow cytometry, while miRNA and protein profiles were assessed via next-generation sequencing and mass spectrometry.

LanguageEnglish

CategoryResearch & Report

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Pages9

Reading Time23 min

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Abstract

Background

Methods

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Introduction

BJS Open, 2025, zrae139 [https://doi.org/10.1093/bjsopen/zrae139](https://doi.org/10.1093/bjsopen/zrae139) Original Article Circulating large extracellular vesicles as diagnostic biomarkers of indeterminate thyroid nodules: multi-platform omics analysis Nada M. Ahmed1,2 , Mohammad M. R. Eddama3,* , Kevin Beatson3 , Rijan Gurung3, Jigisha Patel3, Georges Iskandar4, Alaa Abdel-Salam5 , Abdullah Al-Omar5, Richard Cohen3, Tarek Abdel-Aziz5 and Lucie Clapp1 1Institute of Cardiovascular Sciences, University College London, London, UK 2Pathology Department, Alexandria University, Alexandria, Egypt 3Department of Surgical Biotechnology, Division of Surgery and Interventional Science, University College London, London, UK 4Department of Anaesthesia and Perioperative Medicine, University College London Hospitals, London, UK 5Endocrine Surgery Unit, University College London Hospitals, London, UK *Correspondence to: Mohammad M. R. Eddama, Research Department of Surgical Biotechnology, University College London, GI Services, Ground Floor, 250 Euston Road, London NW1 2PG, UK ([e-mail: m.eddama.12@ucl.ac.uk](e-mail: m.eddama.12@ucl.ac.uk)) Abstract Background: While most thyroid nodules are benign, 7–15% are malignant. Patients with indeterminate thyroid nodules (specifically Bethesda IV/Thy3f) often undergo diagnostic hemithyroidectomy to reach a diagnosis on final histology. The aim of this study was to assess the feasibility of circulating large extracellular vesicles as diagnostic biomarkers in patients presenting with Thy3f thyroid nodules. Methods: This was a two-gate diagnostic accuracy study; patients with Thy3f thyroid nodules were age, sex and body mass index matched to healthy individuals. Final histology confirmed benign and malignant diagnoses. Plasma large extracellular vesicle counts were quantified using flow cytometry. Large extracellular vesicle microRNA and protein profiles were identified using next generation sequencing and mass spectrometry, respectively. Results: A total of 42 patients with Thy3f nodules (22 with cancer, 20 with non-cancer diagnosis) and 16 healthy controls were included. Total large extracellular vesicle concentrations and the concentrations of extracellular vesicles expressing epithelial cell adhesion molecule and the cancer markers atypical chemokine receptor type 7, extracellular matrix metalloproteinase inducer and syndecan-4 were significantly higher in patients with Thy3f nodules (cancer and non-cancer) compared with healthy individuals. In patients with cancerous versus non-cancer Thy3f nodules, one microRNA was upregulated: mir-195–3p (P < 0.001) . Five were downregulated: mir-3176 (P < 0.001), mir-205-5p (P < 0.001), novel-hsa-mir-208-3p (P < 0.001), mir-3529-3p (P = 0.01) and let-7i-3p (P = 0.02) . Furthermore, three large extracellular vesicle proteins (kallikrein-related peptidase11 (KLK11) (P = 0.001), alpha-1-acid glycoprotein 2 (A1AG2) (P <0.001) and small integral membrane protein 1 (SMIM1) (P = 0.04)) were significantly upregulated, while 20 large extracellular vesicle proteins were significantly downregulated (most downregulated: chemokine (C-X-C motif) ligand 7 (CXCL7), tubulin beta chain 1 (TBB1), binding immunoglobulin protein (BIP) and actinin alpha 1 (ACTN1) (P < 0.001)) in cancerous compared with non-cancer Thy3f nodules. Conclusion: Circulating large extracellular vesicle miRNA and protein profiles have a high diagnostic value to discriminate between benign and malignant nodules for patients with Thy3f cytology. Further validation for clinical performance will be needed. Introduction Thyroid cancer incidence has increased three-fold over the past four decades and now has the seventh highest cancer incidence globally. Thyroid nodules are largely benign, however, 7–15% will be malignant depending on factors such as age, sex, radiation exposure and family history1. Clinical assessment, imaging and cytology can establish a diagnosis in approximate

FAQs

Why are indeterminate thyroid nodules (Thy3f/Bethesda IV) clinically challenging?

Most nodules are benign, but 7–15% are malignant, and Thy3f carries an estimated malignancy risk of 15–30%. Current diagnostic hemithyroidectomy is used to confirm histology, creating need for better non-invasive tests.

How were circulating large extracellular vesicles measured in this study?

Plasma large extracellular vesicle concentrations were quantified using flow cytometry. miRNA profiles were identified with next-generation sequencing, and protein profiles were assessed using mass spectrometry.

What key biomarker differences were found between Thy3f nodules and healthy controls?

Total large extracellular vesicle concentrations were higher in Thy3f patients. Vesicles expressing epithelial cell adhesion molecule and several cancer-related markers showed significantly increased concentrations versus healthy individuals.

What was the study’s main conclusion for diagnostic use?

Circulating large extracellular vesicle miRNA and protein profiles demonstrated high diagnostic value to distinguish benign from malignant Thy3f nodules, with further validation required for clinical performance.

Sep 21, 2026

Uploaded by Theodora