TET2 deficiency inhibits mesoderm and hematopoietic differentiation in human embryonic stem cells.
Ten-eleven-translocation 2 (TET2) belongs to the TET protein family that catalyzes the conversion of 5-methylcytosine into 5-hydroxymethylcytosine and plays a central role in normal and malignant adult hematopoiesis. Yet the role of TET2 in human hematopoietic development remains largely unknown. Here, we show that TET2 expression is low in human embryonic stem cell (ESC) lines and increases during hematopoietic differentiation. shRNA-mediated TET2 knockdown had no effect on the pluripotency of various ESCs. However, it skewed their differentiation into neuroectoderm at the expense of endoderm and mesoderm both in vitro and in vivo. These effects were rescued by reintroducing the targeted TET2 protein. Moreover, TET2-driven differentiation was dependent on NANOG transcriptional factor. Indeed, TET2 bound to NANOG promoter and in TET2-deficient cells the methylation of the NANOG promoter correlated with a decreased in NANOG expression. The altered differentiation resulting from TET2 knockdown in ESCs led to a decrease in both the number and the cloning capacities of hematopoietic progenitors. These defects were due to an increased apoptosis and an altered gene expression profile, including abnormal expression of neuronal genes. Intriguingly, when TET2 was knockdown in hematopoietic cells, it increased hematopoietic development. In conclusion, our work suggests that TET2 is involved in different stages of human embryonic development, including induction of the mesoderm and hematopoietic differentiation.
Institut National de la Santé et de la Recherche Médicale, UMR 1009, Laboratory of Excellence GR-Ex, 114 rue Edouard Vaillant, Villejuif, Paris, France; Institut Gustave Roussy, Villejuif, Paris, France; Université Paris Sud 11, Orsay, France.
Thierry
T
Langlois
Barbara
B
da Costa Reis Monte-Mor
Gaëlle
G
Lenglet
Nathalie
N
Droin
Caroline
C
Marty
Jean-Pierre
JP
Le Couédic
Carole
C
Almire
Nathalie
N
Auger
Thomas
T
Mercher
François
F
Delhommeau
Jesper
J
Christensen
Kristian
K
Helin
Najet
N
Debili
François
F
Fuks
Olivier A
OA
Bernard
Eric
E
Solary
William
W
Vainchenker
Isabelle
I
Plo
D015153:Blotting, Western; D002454:Cell Differentiation / Q000502:physiology*; D002460:Cell Line; D047369:Chromatin Immunoprecipitation; D004268:DNA-Binding Proteins / Q000378:metabolism*; D049308:Dioxygenases; D053595:Embryonic Stem Cells / Q000166:cytology*; D005434:Flow Cytometry; D055785:Gene Knockdown Techniques; D006410:Hematopoiesis / Q000502:physiology*; D006412:Hematopoietic Stem Cells / Q000166:cytology*; D018398:Homeodomain Proteins / Q000378:metabolism; D006801:Humans; D008648:Mesoderm / Q000166:cytology / Q000378:metabolism; D000071317:Nanog Homeobox Protein; D011518:Proto-Oncogene Proteins / Q000378:metabolism*; D020133:Reverse Transcriptase Polymerase Chain Reaction
D004268:DNA-Binding Proteins; D018398:Homeodomain Proteins; C486780:NANOG protein, human; D000071317:Nanog Homeobox Protein; D011518:Proto-Oncogene Proteins; D049308:Dioxygenases; C541081:TET2 protein, human
Hematopoiesis; NANOG; TET2; hES