Thyroid Progenitors Are Robustly Derived from Embryonic Stem Cells through Transient, Developmental Stage-Specific Overexpression of Nkx2-1.
The 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.
Center for Regenerative Medicine, Boston Medical Center and Boston University, 670 Albany Street, 2nd Floor CReM, Boston, MA 02118, USA; The Pulmonary Center, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
Keri
K
Dame
Center for Regenerative Medicine, Boston Medical Center and Boston University, 670 Albany Street, 2nd Floor CReM, Boston, MA 02118, USA; The Pulmonary Center, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
Steven
S
Cincotta
Department of Physics, Boston University, Boston, MA 02215, USA.
Alex H
AH
Lang
Center for Regenerative Medicine, Boston Medical Center and Boston University, 670 Albany Street, 2nd Floor CReM, Boston, MA 02118, USA; The Pulmonary Center, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
Reeti M
RM
Sanghrajka
Section of Computational Biomedicine, Boston University School of Medicine, Boston, MA 02118, USA.
Liye
L
Zhang
Center for Regenerative Medicine, Boston Medical Center and Boston University, 670 Albany Street, 2nd Floor CReM, Boston, MA 02118, USA; Division of Endocrinology, Diabetes and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA.
Jinyoung
J
Choi
The Pulmonary Center, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
Letty
L
Kwok
The Pulmonary Center, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
Talitha
T
Wilson
Chair of Bioinformatics Research Group, Boku University, 1190 Vienna, Austria.
Maciej M
MM
Kańduła
Section of Computational Biomedicine, Boston University School of Medicine, Boston, MA 02118, USA.
Stefano
S
Monti
Division of Endocrinology, Diabetes and Metabolism, Beth Israel Deaconess Medical Center, Harvard Medical School, Boston, MA 02215, USA.
Anthony N
AN
Hollenberg
Department of Physics, Boston University, Boston, MA 02215, USA.
Pankaj
P
Mehta
Center for Regenerative Medicine, Boston Medical Center and Boston University, 670 Albany Street, 2nd Floor CReM, Boston, MA 02118, USA; The Pulmonary Center, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA.
Darrell N
DN
Kotton
Center for Regenerative Medicine, Boston Medical Center and Boston University, 670 Albany Street, 2nd Floor CReM, Boston, MA 02118, USA; The Pulmonary Center, Department of Medicine, Boston University School of Medicine, Boston, MA 02118, USA. Electronic address: laertis@bu.edu.
Laertis
L
Ikonomou
D000818:Animals; D055415:Bone Morphogenetic Protein 4 / Q000378:metabolism; D002454:Cell Differentiation / Q000235:genetics*; D002460:Cell Line; D019070:Cell Lineage / Q000235:genetics; D016000:Cluster Analysis; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism*; D016222:Fibroblast Growth Factor 2 / Q000378:metabolism; D015870:Gene Expression; D020869:Gene Expression Profiling; D018507:Gene Expression Regulation, Developmental; D055106:Genome-Wide Association Study; D051379:Mice; D015398:Signal Transduction; D013234:Stem Cells / Q000166:cytology* / Q000378:metabolism*; D013961:Thyroid Gland / Q000166:cytology* / Q000196:embryology / Q000378:metabolism*; D000074482:Thyroid Nuclear Factor 1 / Q000235:genetics*; D019076:Transgenes
D055415:Bone Morphogenetic Protein 4; C000614414:Nkx2-1 protein, mouse; D000074482:Thyroid Nuclear Factor 1; D016222:Fibroblast Growth Factor 2
Nkx2-1; directed differentiation; foregut; inducible; mouse embryonic stem cell; overexpression; pluripotent stem cell; progenitor specification; thyroid development; transcription factor