Thyroid Progenitors Are Robustly Derived from Embryonic Stem Cells through Transient, Developmental Stage-Specific Overexpression of Nkx2-1

Abstract

SummaryThe clinical importance of anterior foregut endoderm (AFE) derivatives, such as thyrocytes, has led to intense research efforts for their derivation through directed differentiation of pluripotent stem cells (PSCs). Here, we identify transient overexpression of the transcription factor (TF) NKX2-1 as a powerful inductive signal for the robust derivation of thyrocyte-like cells from mouse PSC-derived AFE. This effect is highly developmental stage specific and dependent on FOXA2 expression levels and precise modulation of BMP and FGF signaling. The majority of the resulting cells express thyroid TFs (Nkx2-1, Pax8, Foxe1, Hhex) and thyroid hormone synthesis-related genes (Tg, Tpo, Nis, Iyd) at levels similar to adult mouse thyroid and give rise to functional follicle-like epithelial structures in Matrigel culture. Our findings demonstrate that NKX2-1 overexpression converts AFE to thyroid epithelium in a developmental time-sensitive manner and suggest a general methodology for manipulation of cell-fate decisions of developmental intermediates.

Pluripotent stem cell differentiation reveals distinct developmental pathways regulating lung versus thyroid lineage specification

“…4E). Sftpb was also enriched in this cluster, a finding in keeping with the reported expression of this transcript in primary, as well as ESC-derived, thyroid epithelial cells (www.gtexportal.org/home/gene/SFTPB and Dame et al, 2017).…”

“…We have previously demonstrated that, following thyroid-lineage specification, 3D culture outgrowth conditions augment subsequent thyroid epithelial gene expression and allow spontaneous self-organization of Nkx2-1 + cells into epithelial monolayered structures that resemble primary thyroid follicular epithelium. Importantly, these PSC-derived thyroid follicles are capable of sequestering thyroglobulin and initiating thyroid hormone biogenesis both in vitro and after transplantation in vivo (Kurmann et al, 2015;Dame et al, 2017). We verified that Nkx2-1 + cells specified with BMP4+FGF2 formed thyroid follicular epithelia ( Fig.…”

Development of the thyroid gland

“…In addition, the instructive signals that delineate and distinguish thyroid and lung lineages are as yet unresolved, as is the fundamental question of whether these lineages have an ancestral and common endoderm progenitor. Of note, it was recently shown that Nkx2-1 alone can efficiently induce thyroid progenitors in a Foxa2-negative subpopulation of pre-patterned anterior endoderm derived from mouse ESCs (Dame et al, 2017). It is known that Foxa2, which is absolutely required for endoderm formation (Ang et al, 1993;Monaghan et al, 1993), is downregulated specifically in the mouse thyroid bud as compared with the adjacent endoderm and lung bud .…”

Maternal smoking and high BMI disrupt thyroid gland development

“…It is possible that the developmental changes in FOXA2 and FOXE1 expression control thyroid folliculogenesis and that deviations seen in the female fetuses from mothers of BMI ≥ 25 contributed to the perturbed thyroid folliculogenesis. The FOXA2 transcription factor is expressed in the C cell precursors of the mouse fetal thyroid, and it may modulate the expression of TTF1 while also playing a role in thyroid regeneration [41][42][43]. FOXA2 distribution in the human thyroid suggests similar developmental functions, with FOXA2 staining sometimes resembling the distribution of calcitonin-positive C-cells ( Fig.…”