Identification of soluble tissue‑derived biomarkers from human thyroid tissue explants maintained on a microfluidic device

Abstract

Although a large cohort of potential biomarkers for thyroid cancer aggressiveness have been tested in various formats in recent years, to the best of our knowledge, thyroglobulin and calcitonin remain the only two established biomarkers associated with thyroid cancer management. Our group has recently validated a novel means of maintaining live, human ex vivo thyroid tissue within a tissue-on-chip format. The present pilot study aimed to interrogate the tissue effluent, containing all the soluble markers released by the tissue samples maintained within the devices' tissue chamber, for the presence of markers potentially associated with thyroid cancer aggressiveness. Culture effluent from tissue samples harvested from 19 individual patients who had undergone thyroidectomy for the treatment of suspected thyroid cancer was assessed, first using a proteome profiler™ angiogenesis array kit. Patients were subcategorised as 'aggressive' if they possessed a minimum of N1b level metastases, whilst 'non-aggressive' samples were T3 or lower without evidence of multifocality; and contralateral healthy thyroid tissue was harvested for comparative studies. Levels of Serpin-F1, vascular endothelial growth factor, Thrombospondin-1 and chemokine (C-C motif) ligand were significantly altered and, thus, were further investigated using ELISA to allow for quantitative analysis. The concentration of serpin-F1 was significantly increased in the effluent of aggressive thyroid cancer tissue when compared with levels released by both non-aggressive and benign samples. The present study demonstrated the usability of microfluidic technology for the analysis of the ex vivo tissue secretome in order to identify novel biomarkers.

Investigating the effects of arginine methylation inhibitors on microdissected brain tumour biopsies maintained in a miniaturised perfusion system

“…These are well-known patterns that indicate that cell plasma membranes were intact during tissue incubation in the device. 24,[26][27][28][70][71][72] Secondly, we showed that the cytokines released by GBM tissues in the device remained the same for the duration of the experiment. We assayed the levels of 105 cytokines in the effluents corresponding to days 2, 4 and 8 of incubation.…”

“…These are well-known patterns that indicate that cell plasma membranes were intact during tissue incubation in the device. 24,26–28,70–72…”

“…18 Work in this field has included the maintenance and investigation of a range of normal and diseased tissues, e.g. ovarian cancer, 19 breast cancer, 20 lung cancer, 21,22 rectal cancer, 23 thyroid, 24 head and neck [25][26][27][28] and brain tissues. [29][30][31][32] Recently, our group has reported the study of 128 GBM biopsies from 33 patients, using a first-generation brain tumour chip.…”

Development of a dual-flow tissue perfusion device for modeling the gastrointestinal tract–brain axis

“…We have not found sensitivity to be an issue with cells and tissue maintained in other devices we have developed; preliminary investigations confirmed that IL-6 was reliably detected (data not shown). Similar devices used by Riley et al (2021) also successfully identified a panel of factors in effluent using a Proteome Profiler (Biotechne, Abingdon, Oxford). If biomarker release was determined to be too low it is possible to concentrate the effluent, however there are limitations to this as additives to cell medium such as serum can interfere with any subsequent assays once concentrated.…”