Ahmed Nada M, Eddama Mohammad M R, Beatson Kevin, Gurung Rijan, Patel Jigisha, Iskandar Georges, Abdel-Salam Alaa, Al-Omar Abdullah, Cohen Richard, Abdel-Aziz Tarek, Clapp Lucie
Institute of Cardiovascular Sciences, University College London, London, UK.
Pathology Department, Alexandria University, Alexandria, Egypt.
BJS Open. 2024 Dec 30;9(1). doi: 10.1093/bjsopen/zrae139.
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.
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.
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.
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.
虽然大多数甲状腺结节是良性的，但7%-15%是恶性的。甲状腺结节性质不确定的患者（特别是贝塞斯达IV类/Thy3f）通常会接受诊断性半甲状腺切除术，以通过最终组织学检查得出诊断结果。本研究的目的是评估循环大细胞外囊泡作为Thy3f甲状腺结节患者诊断生物标志物的可行性。
这是一项双门诊断准确性研究；Thy3f甲状腺结节患者在年龄、性别和体重指数方面与健康个体相匹配。最终组织学检查证实了良性和恶性诊断。使用流式细胞术对血浆中大细胞外囊泡数量进行定量。分别使用下一代测序和质谱法鉴定大细胞外囊泡的微小RNA和蛋白质谱。
共纳入42例Thy3f结节患者（22例为癌症，20例为非癌症诊断）和16名健康对照者。与健康个体相比，Thy3f结节患者（癌症和非癌症）的大细胞外囊泡总浓度以及表达上皮细胞粘附分子和癌症标志物非典型趋化因子受体7型、细胞外基质金属蛋白酶诱导剂和多配体蛋白聚糖-4的细胞外囊泡浓度显著更高。在患有癌症的Thy3f结节与非癌症的Thy3f结节患者中，一种微小RNA上调：mir-195-3p（P<0.001）。五种微小RNA下调：mir-3176（P<0.001）、mir-205-5p（P<0.001）、新型-hsa-mir-208-3p（P<0.001）、mir-3529-3p（P=0.01）和let-7i-3p（P=0.02）。此外，三种大细胞外囊泡蛋白（激肽释放酶相关肽酶11（KLK11）（P=0.001）、α-1-酸性糖蛋白2（A1AG2）（P<0.001）和小整合膜蛋白1（SMIM1）（P=0.04））显著上调，而与非癌症的Thy3f结节相比，20种大细胞外囊泡蛋白在癌症结节中显著下调（下调最多的是趋化因子（C-X-C基序）配体7（CXCL7）、微管蛋白β链1（TBB1）、结合免疫球蛋白蛋白（BIP）和辅肌动蛋白α1（ACTN1）（P<0.001））。
循环大细胞外囊泡微小RNA和蛋白质谱对于鉴别Thy3f细胞学患者的良性和恶性结节具有较高的诊断价值。还需要对临床性能进行进一步验证。
