Generation and Differentiation of Adult Tissue-Derived Human Thyroid Organoids

Abstract

Total thyroidectomy as part of thyroid cancer treatment results in hypothyroidism requiring lifelong daily thyroid hormone replacement. Unbalanced hormone levels result in persistent complaints such as fatigue, constipation, and weight increase. Therefore, we aimed to investigate a patient-derived thyroid organoid model with the potential to regenerate the thyroid gland. Murine and human thyroidderived cells were cultured as organoids capable of self-renewal and which expressed proliferation and putative stem cell and thyroid characteristics, without a change in the expression of thyroid tumor-related genes. These organoids formed thyroid-tissue-resembling structures in culture. (Xeno-)transplantation of 600,000 dispersed organoid cells underneath the kidney capsule of a hypothyroid mouse model resulted in the generation of hormone-producing thyroid-resembling follicles. This study provides evidence that thyroid-lineagespecific cells can form organoids that are able to self-renew and differentiate into functional thyroid tissue. Subsequent (xeno-)transplantation of these thyroid organoids demonstrates a proof of principle for functional miniature gland formation.

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

“…These three-dimensional cultures are representative in vitro model systems, which recapitulate human physiology (11). While we were finalizing the current study, Coppes and colleagues reported a first description of transplantable human (and mouse) thyroid organoids (12). In this study, we report similar findings and apply thyroid follicular cell organoids (TFCOs) to model Graves' disease using patient sera.…”

“…Since we observed a gradually increasing growth rate from passage 5 onwards, we hypothesized that such stem cell markers would become evident when comparing tissue to early passage TFCOs and would increase in expression from early to late passage organoids. No significant change in expression was identified in the putative stem cell markers SCA1 (17,18), ABCG2 (17,19), FUT4 (12), GATA4 (20), HNF4A (20), NANOG (21), OCT4 (12,17,(20)(21)(22), and HHEX (15,23) in human TFCOs (SI Appendix, Fig. S3 A-J).…”

“…S3 A-J). Increased expression was noted for NGFR and FOXE1 (TTF2) (24), while SOX2 (12,22) was enriched in late passage human TFCOs (SI Appendix, Fig. S3 K-M).…”

“…The optimized protocol allowed the generation of 10 human organoid lines from 12 provided donor tissue samples (±83%). Several other groups have recently reported similar thyroid organoids derived from mouse (34), rat (35), and human tissue (12). Related approaches have resulted in the establishment of human thyroid cancer organoids (36,37).…”

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

“…Human embryonic stem cell (ESC)-based protocols have been developed and led to the generation of several types of human organoids that include brain, intestine, stomach, liver, kidney, lung, endometrium, prostate, pancreas, and retina 8,9 . With regard to the thyroid, murine ESC-derived organoids have been shown to recapitulate in vitro the developmental stages of the thyroid gland with the ability to produce TH in vitro and in vivo after transplantation to mice with ablated thyroid glands [11][12][13][14][15] .…”