A novel microfluidic device capable of maintaining functional thyroid carcinoma specimens ex vivo provides a new drug screening platform

Riley, Andrew; Green, Victoria; Cheah, Ramsah; McKenzie, Gordon; Karsai, Laszlo; England, James; Greenman, John

Abstract

© 2019 The Author(s). Background: Though the management of malignancies has improved vastly in recent years, many treatment options lack the desired efficacy and fail to adequately augment patient morbidity and mortality. It is increasingly clear that patient response to therapy is unique to each individual, necessitating personalised, or 'precision' medical care. This demand extends to thyroid cancer; ~ 10% patients fail to respond to radioiodine treatment due to loss of phenotypic differentiation, exposing the patient to unnecessary ionising radiation, as well as delaying treatment with alternative therapies. Methods: Human thyroid tissue (n = 23, malignant and benign) was live-sliced (5 mm diameter × 350-500 μm thickness) then analysed or incorporated into a microfluidic culture device for 96 h (37 °C). Successful maintenance of tissue was verified by histological (H&E), flow cytometric propidium iodide or trypan blue uptake, immunohistochemical (Ki67 detection/ BrdU incorporation) and functional analysis (thyroxine [T4] output) in addition to analysis of culture effluent for the cell death markers lactate dehydrogenase (LDH) and dead-cell protease (DCP). Apoptosis was investigated by Terminal deoxynucleotidyl transferase dUTP nick end labelling (TUNEL). Differentiation was assessed by evaluation of thyroid transcription factor (TTF1) and sodium iodide symporter (NIS) expression (western blotting). Results: Maintenance of gross tissue architecture was observed. Analysis of dissociated primary thyroid cells using flow cytometry both prior to and post culture demonstrated no significant change in the proportion of viable cells. LDH and DCP release from on-chip thyroid tissue indicated that after an initial raised level of release, signifying cellular damage, detectable levels dropped markedly. A significant increase in apoptosis (p < 0.01) was observed after tissue was perfused with etoposide and JNK inhibitor, but not in control tissue incubated for the same time period. No significant difference in Ki-67 positivity or TTF1/NIS expression was detected between fresh and post-culture thyroid tissue samples, moreover BrdU positive nuclei indicated on-chip cellular proliferation. Cultured thyroid explants were functionally viable as determined by production of T4 throughout the culture period. Conclusions: The described microfluidic platform can maintain the viability of thyroid tissue slices ex vivo for a minimum of four days, providing a platform for the assessment of thyroid tissue radioiodine sensitivity/adjuvant therapies in real time.

Citation

Riley, A., Green, V., Cheah, R., McKenzie, G., Karsai, L., England, J., & Greenman, J. (2019). A novel microfluidic device capable of maintaining functional thyroid carcinoma specimens ex vivo provides a new drug screening platform. BMC Cancer, 19(1), Article 259. https://doi.org/10.1186/s12885-019-5465-z

• Journal Article Type: Article
• Acceptance Date: Mar 12, 2019
• Online Publication Date: Mar 22, 2019
• Publication Date: Mar 22, 2019
• Deposit Date: Apr 24, 2019
• Publicly Available Date: Apr 24, 2019
• Journal: BMC Cancer
• Print ISSN: 1471-2407
• Electronic ISSN: 1471-2407
• Publisher: BioMed Central
• Peer Reviewed: Peer Reviewed
• Volume: 19
• Issue: 1
• Article Number: 259
• DOI: https://doi.org/10.1186/s12885-019-5465-z
• Keywords: Thyroid gland; Microfluidics; Viability; De-differentiation; Radioiodine therapy
• Public URL: https://hull-repository.worktribe.com/output/1373578
• Publisher URL: https://bmccancer.biomedcentral.com/articles/10.1186/s12885-019-5465-z
• Contract Date: Apr 24, 2019