Can Tissue-On-Chip Technology provide a platform for investigating differences in Extracellular Vesicle miRNA content between Thyroid Pathologies?

Haigh, Thomas

Abstract

This study uses an established method to maintain human thyroid tissue ex vivo on a tissue-on-chip device, allowing for the collection, isolation and interrogation of the small extracellular vesicles (sEVs) released directly from thyroid tissue. The size and concentration of the sEVs from the thyroid tissue on chip effluent was analysed using
nanoparticle tracking analysis (NTA) (n=11) along with histoarchitectural integrity following both pre and post incubation on chip (n=16). sEVs were analysed for differences in miRNA levels released from euthyroid multinodular goitre (EMG), Graves’ disease (GD) and papillary thyroid cancer (PTC), using miRNA sequencing (miRNASeq) and quantitative reverse transcriptase polymerase chain reaction (RT-qPCR) to identify potential biomarkers of disease. Thyroid biopsies from patients with EMG (n = 5), GD (n= 5) and PTC (n = 5) were perfused with medium containing sEV-depleted serum for 6 days on the tissue-on-chip device. All the effluents were collected and ultracentrifuged to isolate sEVs; miRNA was extracted and sequenced. Out of the 15 samples, 14 passed the quality control and miRNASeq analysis detected significantly higher expression of miR-375-3p, miR-7-5p, miR-382-5p and miR-127-3p in the sEVs isolated from GD tissue compared to those from EMG (false discovery rate; FDR < 0.05). Similarly, miR-375-3p and miR-7-5p were also detected at a higher level in the GD tissue sEVs compared to the PTC tissue sEVs (FDR < 0.05). No significant differences were observed between sEV miRNA from PTC vs. EMG. These results were supported by RT-qPCR. The novel findings demonstrate that the tissue-on-chip technology is a robust method for isolating sEVs directly from the tissue of interest. This has permitted the identification of four miRNAs, which through further investigation could be used as biomarkers or therapeutic targets within thyroid disease. Further works will look to validate these findings in patient serum samples with the long-term aim of developing a non-invasive liquid biopsy to help diagnose and manage thyroid disease.

Citation

Haigh, T. (2025). Can Tissue-On-Chip Technology provide a platform for investigating differences in Extracellular Vesicle miRNA content between Thyroid Pathologies?. (Thesis). Hull York Medical School, University of Hull and University of York. https://hull-repository.worktribe.com/output/5182708

• Thesis Type: Thesis
• Deposit Date: May 23, 2025
• Publicly Available Date: Jun 24, 2025
• Keywords: Biomedical sciences
• Public URL: https://hull-repository.worktribe.com/output/5182708
• Additional Information: Biomedical Sciences; Hull York Medical School; University of Hull and University of York
• Award Date: May 1, 2025