Circulating MicroRNA Profiles as Potential Biomarkers for differentiated thyroid cancer recurrence

Context Circulating microRNAs (miRNAs) are emerging biomarkers of thyroid cancer. Objective This study sought to identify the profile of circulating miRNAs and its response to human recombinant TSH (rhTSH) in thyroid cancer patients with recurrent/persistent disease. Methods We obtained serum samples from 30 patients with differentiated thyroid cancer, 14 with recurrent/persistent disease and 16 with complete remission. We used next generation sequencing to define the miRnomes along with a comprehensive qPCR validation using two different platforms. We made a transversal study by comparing serum miRNA profiles of patients with or without recurrent/persistent disease and a longitudinal study looking at differences before and after rhTSH stimulation. Selected miRNAs were then studied in human thyroid cancer cell lines TPC-1, FTC-133 and OCUT-2 in response to TSH stimulation. Results We could not demonstrate any consistent differences in serum profiles of known miRNAs between patients with and without recurrent/persistent disease or before and after rhTSH stimulation. However, our sequencing data revealed two putative novel miRNAs that rise with rhTSH stimulation in the serums of patients with recurrent/persistent disease. We further confirmed by qPCR the upregulation of these putative miRNAs both in serums and in TSH-stimulated cells. We also show miRNAs that are good candidates for housekeeping genes in the serum of patients independently of the levels of TSH. Conclusions The present study does not provide evidence that known miRNAs can be used as circulating markers for recurrence of thyroid cancer. However, we suggest that novel miRNA molecules may be related to thyroid cancer pathogenesis.

Adult mouse and human organoids derived from thyroid follicular cells and modeling of Graves’ hyperthyroidism

The thyroid maintains systemic homeostasis by regulating serum thyroid hormone concentrations. Here we report the establishment of three-dimensional (3D) organoids from adult thyroid tissue representing murine and human thyroid follicular cells (TFCs). The TFC organoids (TFCOs) harbor the complete machinery of hormone production as visualized by the presence of colloid in the lumen and by the presence of essential transporters and enzymes in the polarized epithelial cells that surround a central lumen. Both the established murine as human thyroid organoids express canonical thyroid markers PAX8 and NKX2.1, while the thyroid hormone precursor thyroglobulin is expressed at comparable levels to tissue. Single-cell RNA sequencing and transmission electron microscopy confirm that TFCOs phenocopy primary thyroid tissue. Thyroid hormones are readily detectable in conditioned medium of human TFCOs. We show clinically relevant responses (increased proliferation and hormone secretion) of human TFCOs toward a panel of Graves’ disease patient sera, demonstrating that organoids can model human autoimmune disease.

To Study the Deteriorating Effects of Lithium Carbonate on Thyroid Gland - A Study in Albino Rats

Background: The human thyroid gland is located in the front of neck. It consists of two lobes. The two lobes are joined with each other by isthmus. The mood stabilizer Lithium Caronate has deleterious effects on the thyroid gland. Aim: To observe and report the data of the harmful effect of Lithium on the weight changes of thyroid gland. Methods: Sixteen rats were selected for this experimental study. The rodents were divided into two groups. Group A comprised of eight animals which were given laboratory diet, Group B contained eight albinos who were given Tablet Lithium Carbonate in powder form at a dose of 60 mg/day for four weeks. After completion of the study time animals were sacrificed and thyroid gland weight were recorded and compared in both groups. Results: The results in both groups were recorded and compared .It was reported that Group B animals had a highly significantly decreased thyroid weight after four weeks Lithium ingestion than Group A control group. Conclusion: The results of our study concluded that Lithium Carbonate damages thyroid glandular tissue and causes its weight to decline. Key words: Thyroid gland, Isthmus, deteriorating

THYROID FUNCTION OF RATS WITH EXPERIMENTAL AIT AT AN EARLY PERIOD AFTER THE INJECTION OF BIOPREPARATIONS OF CORD BLOOD

The development of new methods for the treatment of autoimmune thyroid disease is an actual problem of endocrinology. The aim of research was studying of the cell-free effect biological product of cord blood with immunomodulatory properties "Cryocell-Cryocord " on rats’ thyroid function with AIT simulated at an early period of the study. Simulation of autoimmune thyroiditis was carried out by immunization with a suboperatively isolated human thyroid antigen in combination completing Freund's adjuvant on 30 adult male rats weighing 130 - 150 g. Correction was carried out by parenteral ingection of the "Cryocell-Cryocord". Hormonal research (determination of the content of thyroid hormones and antibodies to thyroglobulin (TG-AT) were effected by using standard test kits for enzyme-linked immunosorbent assay at early stages after exposure (7 days, 1 month). Statistical maintained of the results was carried out using the methods of variation statistics. It was used normal distribution to unpaired Student's test to compare indicators. The difference was considered significant at p<0.05. It has been shown the biological preparation of cord blood "Cryocell - Cryocord" has a positive effect with induced autoimmune thyroiditis. It has been noted the normalizing result of the effected correction on the thyroid function of animals in the early period of the studying. It has been proved after the introduction of the biological preparation "Cryocell - Cryocord" in animals with experimental autoimmune thyroiditis observed the inhibition of the manifestations of autoimmune aggression and the normalization of the functional activity of the thyroid gland. These investigations can be the foundation for the development of a new approach to the treatment of patients.

Transplantable human thyroid organoids generated from embryonic stem cells to rescue hypothyroidism

The function of the thyroid gland is to capture iodide in order to synthesize hormones that act on almost all tissues and are essential for normal growth and metabolism. Low plasma levels of thyroid hormones lead to hypothyroidism, which is one of the most common disorder in humans which is not always satisfactorily treated by lifelong hormone replacement. Therefore, in addition to the lack of in vitro tractable models to study human thyroid development, differentiation and maturation, there is a need for new therapeutic approaches that involve replacement of thyroid tissue responsive to changing demands for thyroid hormone. Here we report the first transplantable thyroid organoids derived from human embryonic stem cells capable of restoring plasma thyroid hormone to athyreotic mice as a proof of concept for future therapeutic development.

Overexpression of PAX8-AS1 Inhibits Malignant Phenotypes of Papillary Thyroid Carcinoma Cells via miR-96-5p/PKN2 Axis

Background. Thyroid carcinoma (THCA) is the most frequent endocrine malignancy. Papillary thyroid carcinoma (PTC) is the major subtype of THCA, accounting for over 80% of all THCA cases. LncRNA PAX8-AS1, a tumor suppressor associated with various human cancers, has been reported to be relevant to the regulation of all sorts of cellular processes. The purpose of this study was to verify the role of PAX8-AS1 in PTC. Methods. Three human PTC cell lines (K1, TPC-1, and IHH4) and one normal human thyroid cell line, Nthy-ori3-1, were used in our study. The expression of genes was detected by qRT-PCR. The bioinformatic analysis and luciferase reporter assay were used to confirm the binding relationship of PAX8-AS1 to miR-96-5p, and the targeting relationship of miR-96-5p to PKN2 was also predicted. Cell proliferation and apoptosis capacities were assessed by MTT and flow cytometry, respectively. EdU assay was used to detect cell proliferation. Western blot assay was employed to examine protein expression. Results. The expression of PAX8-AS1 was decreased in PTC tissues and cells. PAX8-AS1 overexpression inhibited the proliferation of PTC cells and promoted cell apoptosis. In addition, PAX8-AS1 bonds with miR-96-5p, whose downregulation elevated the expression of PKN2 in PTC cells. Importantly, according to the rescue experiments, PKN2 silencing partially reversed the inhibitory effects of PAX8-AS1 expression on PTC cell proliferation and apoptosis. Conclusions. We found that the PAX8-AS1/miR-96-5p/PKN2 axis was closely related to the progression of PTC, which could be a potential target for treating PTC patients.