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

2021 ◽  
Vol 118 (51) ◽  
pp. e2117017118
Author(s):  
Jelte van der Vaart ◽  
Lynn Bosmans ◽  
Stijn F. Sijbesma ◽  
Kèvin Knoops ◽  
Willine J. van de Wetering ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Hormone Secretion ◽  
Thyroid Tissue ◽  
Three Dimensional ◽  
Disease Patient ◽  
Human Thyroid ◽  
Follicular Cells ◽  
Hormone Production ◽  
Transmission Electron ◽  
Thyroid Follicular Cells

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.

scholarly journals Generation of Adult Stem Cell Derived Organoid Cultures From Thyroid Follicular Cells

2021 ◽  
Vol 5 (Supplement_1) ◽  
pp. A850-A851
Author(s):  
Jelte van der Vaart ◽  
Lynn Bosmans ◽  
Hanneke Margo van Santen ◽  
Menno R Vriens ◽  
Hans Clevers
Keyword(s):  
Stem Cell ◽  
Thyroid Hormone ◽  
Culture System ◽  
Adult Stem Cell ◽  
Human Thyroid ◽  
Wild Type ◽  
Follicular Cells ◽  
Hormone Production ◽  
Organoid Culture ◽  
Thyroid Follicular Cells

Abstract The thyroid is essential for maintaining systemic homeostasis by regulating thyroid hormone concentrations in the bloodstream. Due to the limited number of representative model systems, there is limited understanding of fundamental thyroid biology as well as thyroid carcinogenesis. To fill the caveats in the understanding of thyroid cell biology, we aimed to develop an adult stem cell-derived three-dimensional (3D) organoid culture system using murine and human thyroid follicular cells (TFCs). We have succeeded to grow such an organoid culture system that harbours the complete machinery of hormone production visualised by the presence of colloid in the lumen and essential transporters and enzymes in a polarised cell layer. Both the established murine as human thyroid organoids express canonical thyroid markers PAX8 and NKX2.1/TTF1. Moreover, the thyroid hormone precursor thyroglobulin is expressed in both cultures to similar levels as in tissue. Extensive characterisation furthermore identifies known and new biological insights in TFC subclassification, subcellular organisation and hormone production using state-of-the art techniques like single cell RNA sequencing, transmission electron microscopy and genome editing. These 3D in vitro cultures allow for a variety of thyroid-related studies including the progression of wild type cells towards cancer. Additionally, due to the success of generating patient-specific tumour organoids of primary differentiated thyroid carcinoma and metastasis, insights in drug resistance and metastases can be identified. In short, this newly developed organoid culture of murine and human wild type TFCs as well as tumour tissue opens up an extensive area of research that will help understand the drivers for growth and development of thyroid (cancer) cells and enable studies upon drug responsiveness.

pax2.1is required for the development of thyroid follicles in zebrafish

Development ◽  
2002 ◽  
Vol 129 (15) ◽  
pp. 3751-3760 ◽  
Author(s):  
Thomas Wendl ◽  
Klaus Lun ◽  
Marina Mione ◽  
Jack Favor ◽  
Michael Brand ◽  
...  
Keyword(s):  
Thyroid Hormone ◽  
Thyroid Gland ◽  
Marker Gene ◽  
Follicular Cells ◽  
Hormone Production ◽  
Zebrafish Model ◽  
Hormone Synthesis ◽  
Larval Stages ◽  
Thyroid Follicular Cells ◽  
Thyroid Development

The thyroid gland is an organ primarily composed of endoderm-derived follicular cells. Although disturbed embryonic development of the thyroid gland leads to congenital hypothyroidism in humans and mammals, the underlying principles of thyroid organogenesis are largely unknown. In this study, we introduce zebrafish as a model to investigate the molecular and genetic mechanisms that control thyroid development. Marker gene expression suggests that the molecular pathways of early thyroid development are essentially conserved between fish and mammals. However during larval stages, we find both conserved and divergent features of development compared with mammals. A major difference is that in fish, we find evidence for hormone production not only in thyroid follicular cells, but also in an anterior non-follicular group of cells.We show that pax2.1 and pax8, members of the zebrafish pax2/5/8 paralogue group, are expressed in the thyroid primordium. Whereas in mice, only Pax8 has a function during thyroid development, analysis of the zebrafish pax2.1 mutant no isthmus (noi–/–) demonstrates that pax2.1 has a role comparable with mouse Pax8 in differentiation of the thyroid follicular cells. Early steps of thyroid development are normal in noi–/–, but later expression of molecular markers is lost and the formation of follicles fails. Interestingly, the anterior non-follicular site of thyroid hormone production is not affected in noi–/–. Thus, in zebrafish, some remaining thyroid hormone synthesis takes place independent of the pathway leading to thyroid follicle formation. We suggest that the noi–/– mutant serves as a new zebrafish model for hypothyroidism.

scholarly journals Rabbit thyroid extracellular matrix as a 3D bioscaffold for thyroid bioengineering: a preliminary in vitro study

2021 ◽  
Author(s):  
Jie Weng ◽  
Bi Chen ◽  
Mengying Xie ◽  
Xinlong Wan ◽  
Wang Peng ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Three Dimensional ◽  
Biomechanical Properties ◽  
Space Structure ◽  
Thyroid Diseases ◽  
Human Thyroid ◽  
Thyroid Peroxidase ◽  
Follicular Cells ◽  
Thyroid Follicular Cells

Abstract Background: Advances in regenerative medicine technologies have been strongly proposed in the management of thyroid diseases. Mechanistically, the adoption of thyroid bioengineering requires a scaffold that shares a similar three dimensional (3D) space structure, biomechanical properties, protein component, and cytokines to the native extracellular matrix (ECM).Methods: 24 male New Zealand white rabbits were used in this experimental study. The rabbit thyroid glands were decellularized by immersion/agitation decellularization protocol. The 3D thyroid decellularization scaffolds were tested with histological and immunostaining analyses, scanning electron microscopy, DNA quantification, mechanical properties test, cytokine assay and cytotoxicity assays. Meanwhile, the decellularization scaffold were seeded with human thyroid follicular cells, cell proliferation and thyroid peroxidase were determined to explore the biocompatibility in vitro.Results: Notably, through the imaging studies, it was distinctly evident that our protocol intervention minimized cellular materials and maintained the 3D spatial structure, biomechanical properties, ECM composition, and biologic cytokine. Consequently, the decellularization scaffold was seeded with human thyroid follicular cells, thus strongly revealing its potential in reinforcing cell adhesion, proliferation, and preserve important protein expression.Conclusions: The adoption of our protocol to generate a decellularized thyroid scaffold can potentially be utilized in transplantation to manage thyroid diseases through thyroid bioengineering.

scholarly journals Rabbit thyroid extracellular matrix as a 3D bioscaffold for thyroid bioengineering

2020 ◽  
Author(s):  
Jie Weng ◽  
Bi Chen ◽  
Mengying Xie ◽  
Xinlong Wan ◽  
Peng Wang ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Three Dimensional ◽  
Biomechanical Properties ◽  
Space Structure ◽  
Thyroid Diseases ◽  
Human Thyroid ◽  
Imaging Studies ◽  
Follicular Cells ◽  
3D Space ◽  
Thyroid Follicular Cells

Abstract Advances in regenerative medicine technologies have been strongly proposed in the management of thyroid diseases. Mechanistically, the adoption of thyroid bioengineering requires a scaffold that shares a similar three dimensional (3D) space structure, biomechanical properties, protein component, and cytokines to the native extracellular matrix (ECM). Herein, we prepared the 3D thyroid scaffold from the decellularization rabbit thyroid. Notably, through the imaging studies, it was distinctly evident that our protocol intervention minimized cellular materials and maintained the 3D spatial structure, biomechanical properties, ECM composition, and biologic cytokine. Consequently, the decellularization scaffold was seeded with human thyroid follicular cells, thus strongly revealing its potential in reinforcing cell adhesion, proliferation, and preserve important protein expression. Therefore, these findings revealed that the adoption of our protocol to generate a decellularized thyroid scaffold can potentially be utilized in transplantation to manage thyroid diseases through thyroid bioengineering.

scholarly journals SAT-LB78 TSH Is a Negative Modulator of Hippo Transcriptional Effectors

2020 ◽  
Vol 4 (Supplement_1) ◽  
Author(s):  
Celia Fernández-Méndez ◽  
Pilar Santisteban
Keyword(s):  
Thyroid Gland ◽  
Hippo Pathway ◽  
Thyroid Tissue ◽  
Tissue Growth ◽  
Human Thyroid ◽  
In Vivo Model ◽  
Future Research ◽  
Follicular Cells ◽  
Thyroid Follicular Cells ◽  
The Hippo Pathway

Abstract Hippo signaling pathway regulation by hormonal signals acting through G-coupled receptors has been widely described. Modulation of processes such as tissue growth or differentiation by this pathway critically relies on the location and levels of its major effectors: the cofactors YAP/TAZ and the family TEAD of transcription factors. Despite this well-defined regulatory mechanism, little is known about the Hippo pathway in the thyroid gland. Thyrotropin (TSH), main factor for thyroid follicular cells differentiation, plays its role by interacting with its G-protein-coupled receptor (TSHR). High serum TSH levels are associated with hypothyroidism, characterized by a change in thyroid follicle morphology and inflammation of the thyroid gland. This led us to study if TSH could modulate the Hippo pathway.Rat thyroid follicular cells (PCCl3) were treated with TSH and forskolin, an adenylyl cyclase activator. By immunofluorescence and western blot, levels and subcellular location of the Hippo Pathway components were assessed in different conditions. An increase of the Hippo kinase MST1/2 and LATS1/2 was observed after TSH and forskolin treatments, corresponding to a downregulation of the transcriptional mediators of the pathway TAZ, YAP and Tead1. Especially remarkable is the translocation of YAP/TAZ from the nucleus, which involves a decrease in their activity.Next, we validated the results in an in vivo model generating hypothyroidism in 3-month-old male C57BL/6J by adding MMI (2-Mercapto-1-Methylimidazole) and perchlorate (KClO4) to their drinking water. After 2 weeks of treatment, we euthanized the animals, validated higher TSH serum levels and performed analysis of the Hippo components in the thyroid by immunohistochemistry. A reduction in the levels of the Hippo effectors TAZ, YAP and Tead1 was found in the thyroid slices from hypothyroid mice, confirming the in vitro results. In addition, evaluation of a human thyroid tissue microarray, including Hashimoto disease samples, led to a validation of the previously described TSH role.Hereby, we report a crosstalk by which TSH is increasing the kinase axis of the Hippo pathway thus decreasing the activity of its main transcriptional effectors in the nuclei. Future research of the role of these transcriptional effectors will be carry out to discern if their decrease could be associated with the morphology changes linked to hypothyroidism.

scholarly journals Rabbit Thyroid Extracellular Matrix as a 3D Bioscaffold for Thyroid Bioengineering

2020 ◽  
Author(s):  
Jie Weng ◽  
Bi Chen ◽  
Mengying Xie ◽  
Xinlong Wan ◽  
Wang Peng ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Three Dimensional ◽  
Biomechanical Properties ◽  
Space Structure ◽  
Thyroid Diseases ◽  
Human Thyroid ◽  
Thyroid Peroxidase ◽  
Follicular Cells ◽  
3D Space ◽  
Thyroid Follicular Cells

Abstract BackgroundAdvances in regenerative medicine technologies have been strongly proposed in the management of thyroid diseases. Mechanistically, the adoption of thyroid bioengineering requires a scaffold that shares a similar three dimensional (3D) space structure, biomechanical properties, protein component, and cytokines to the native extracellular matrix (ECM).Methods24 male New Zealand white rabbits were used in this experimental study. The rabbit thyroid glands were decellularized by immersion/agitation decellularization protocol. The 3D thyroid decellularization scaffolds were tested with histological and immunostaining analyses, scanning electron microscopy, DNA quantification, mechanical properties test, cytokine assay and cytotoxicity assays. Meanwhile, the decellularization scaffold were seeded with human thyroid follicular cells, cell proliferation and thyroid peroxidase were determined.ResultsNotably, through the imaging studies, it was distinctly evident that our protocol intervention minimized cellular materials and maintained the 3D spatial structure, biomechanical properties, ECM composition, and biologic cytokine. Consequently, the decellularization scaffold was seeded with human thyroid follicular cells, thus strongly revealing its potential in reinforcing cell adhesion, proliferation, and preserve important protein expression.ConclusionsThe adoption of our protocol to generate a decellularized thyroid scaffold can potentially be utilized in transplantation to manage thyroid diseases through thyroid bioengineering.

ADENYL CYCLASE ACTIVITY IN HUMAN THYROID PLASMA MEMBRANES FROM NORMAL HUMAN THYROID TISSUE AND THYROID ADENOMAS

1979 ◽  
Vol 92 (1) ◽  
pp. 95-104 ◽  
Author(s):  
O. Wålinder ◽  
F. A. Karlsson ◽  
P. A. Dahlberg
Keyword(s):  
Thyroid Hormone ◽  
Plasma Membranes ◽  
Thyroid Tissue ◽  
Human Thyroid ◽  
Thyroid Cell ◽  
Hormone Production ◽  
Adenyl Cyclase Activity ◽  
Production Characteristic ◽  
Hormone Synthesis ◽  
Thyroid Adenomas

ABSTRACT The adenylcyclase (AC) activity of crude human thyroid plasma membranes were studied in some detail and conditions for optimal cyclic AMP-production established. Membranes from eight "cold" and two "hot" thyroid adenomas were investigated and compared to membranes from corresponding normal, paranodular tissues. The investigated membranes were found to contain similar basal AC activities, which were stimulated three to five times with TSH and 20–30 times with fluoride. Further, no difference in the TSH-sensitivity of AC from normal and "cold" adenomas could be detected. Thyrotrophin 0.02 mU/ml caused a measurable and 0.15 mU/ml a half-maximal stimulation of the cAMP-production. The data indicate, that disturbances of thyroid hormone production characteristic of thyroid adenomas, are not caused by alterations of the TSH-AC system and suggest that in a thyroid cell the degree of thyroid hormone synthesis do not alter the amount of plasma membrane-bound adenylcyclase.

scholarly journals Notch Signaling Is Involved in Expression of Thyrocyte Differentiation Markers and Is Down-Regulated in Thyroid Tumors

2008 ◽  
Vol 93 (10) ◽  
pp. 4080-4087 ◽  
Author(s):  
E. Ferretti ◽  
E. Tosi ◽  
A. Po ◽  
A. Scipioni ◽  
R. Morisi ◽  
...  
Keyword(s):  
Gene Expression ◽  
Thyroid Cancer ◽  
Notch Signaling ◽  
Notch Pathway ◽  
Human Thyroid ◽  
Thyroid Tumors ◽  
Follicular Cells ◽  
Differentiation Markers ◽  
Thyroid Follicular Cells ◽  
Thyroid Cancer Cells

Context: Notch genes encode receptors for a signaling pathway that regulates cell growth and differentiation in various contexts, but the role of Notch signaling in thyroid follicular cells has never been fully published. Objective: The objective of the study was to characterize the expression of Notch pathway components in thyroid follicular cells and Notch signaling activities in normal and transformed thyrocytes. Design/Setting and Patients: Expression of Notch pathway components and key markers of thyrocyte differentiation was analyzed in murine and human thyroid tissues (normal and tumoral) by quantitative RT-PCR and immunohistochemistry. The effects of Notch overexpression in human thyroid cancer cells and FTRL-5 cells were explored with analysis of gene expression, proliferation assays, and experiments involving transfection of a luciferase reporter construct containing human NIS promoter regions. Results: Notch receptors are expressed during the development of murine thyrocytes, and their expression levels parallel those of thyroid differentiation markers. Notch signaling characterized also normal adult thyrocytes and is regulated by TSH. Notch pathway components are variably expressed in human normal thyroid tissue and thyroid tumors, but expression levels are clearly reduced in undifferentiated tumors. Overexpression of Notch-1 in thyroid cancer cells restores differentiation, reduces cell growth rates, and stimulates NIS expression via a direct action on the NIS promoter. Conclusion: Notch signaling is involved in the determination of thyroid cell fate and is a direct regulator of thyroid-specific gene expression. Its deregulation may contribute to the loss of differentiation associated with thyroid tumorigenesis.

Regulation of ICAM-1 Expression on Human Thyroid Follicular Cells

1991 ◽  
pp. 223-230
Author(s):  
E. Tolosa ◽  
M. Marti ◽  
C. Roura ◽  
A. Lucas ◽  
R. Pujol-Borrell
Keyword(s):  
Human Thyroid ◽  
Follicular Cells ◽  
Thyroid Follicular Cells

In vivo effects of flavonoid EMD 21388 on thyroid hormone secretion and metabolism in rats

1991 ◽  
Vol 261 (2) ◽  
pp. E227-E232 ◽  
Author(s):  
J. P. Schroder-van der Elst ◽  
D. van der Heide ◽  
J. Kohrle
Keyword(s):  
Thyroid Hormone ◽  
Hormone Secretion ◽  
Male Rats ◽  
Intact Male ◽  
Hormone Production ◽  
Iodide Uptake ◽  
Brown Adipose ◽  
In Vivo Effects

In vitro, the synthetic flavonoid EMD 21388 appears to be a potent inhibitor of thyroxine (T4) 5'-deiodinase and diminishes binding of T4 to transthyretin. In this study, in vivo effects of long-term administration of EMD 21388 on thyroid hormone production and metabolism were investigated. Intact male rats received EMD 21388 (20 mumol.kg body wt-1.rat-1.day-1) for 14 days. [125I]T4 and 3,5,3'-[131I]triiodotyronine (T3) were infused continuously and intravenously in a double-isotope protocol for the last 10 and 7 days, respectively. EMD 21388 decreased plasma thyroid hormone concentrations, but thyrotropin levels in plasma and pituitary did not change. Plasma clearance rates for T4 and T3 increased. Thyroidal T4 secretion was diminished, but T3 secretion was elevated. Extrathyroidal T3 production by 5'-deiodination was lower. T4 concentrations were markedly lower in all tissues investigated. Total tissue T3 was lower in brown adipose tissue, brain, cerebellum, and pituitary, tissues that express the type II 5'-deiodinase isozyme due to decreased local T3 production. Most tissues showed increased tissue/plasma ratios for T4 and T3. These results indicate that this flavonoid diminished T4 and increased T3 secretion by the thyroid, probably in analogy with other natural flavonoids, by interference with one or several steps between iodide uptake, organification, and hormone synthesis.

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