Abstract
TET2 converts 5-methylcytosine to 5-hydroxymethylcytosine (5-hmC) in DNA and is frequently mutated in myeloid malignancies, including myeloproliferative neoplasms. Here we show that the level of 5-hmC is decreased in granulocyte DNA from myeloproliferative neoplasm patients with TET2 mutations compared with granulocyte DNA from healthy patients. Inhibition of TET2 by RNA interference decreases 5-hmC levels in both human leukemia cell lines and cord blood CD34(+) cells. These results confirm the enzymatic function of TET2 in human hematopoietic cells. Knockdown of TET2 in cord blood CD34(+) cells skews progenitor differentiation toward the granulomonocytic lineage at the expense of lymphoid and erythroid lineages. In addition, by monitoring in vitro granulomonocytic development we found a decreased granulocytic differentiation and an increase in monocytic cells. Our results indicate that TET2 disruption affects 5-hmC levels in human myeloid cells and participates in the pathogenesis of myeloid malignancies through the disturbance of myeloid differentiation.
Publication types
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Research Support, N.I.H., Extramural
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Research Support, Non-U.S. Gov't
MeSH terms
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5-Methylcytosine / metabolism*
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Cell Line, Tumor
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Cell Lineage
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Colony-Forming Units Assay
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Cytosine / analogs & derivatives*
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Cytosine / biosynthesis
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DNA Methylation / genetics*
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DNA-Binding Proteins / antagonists & inhibitors
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DNA-Binding Proteins / genetics
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DNA-Binding Proteins / physiology*
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Dioxygenases
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Erythropoiesis / genetics*
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Erythropoiesis / physiology
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Fetal Blood / cytology
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Genetic Vectors / genetics
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Granulocytes / metabolism
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Granulocytes / pathology
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Hematopoietic Stem Cells / cytology*
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Humans
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Lentivirus / genetics
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Monocytes / metabolism
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Monocytes / pathology
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Mutation
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Myelopoiesis / genetics*
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Myelopoiesis / physiology
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Proto-Oncogene Proteins / antagonists & inhibitors
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Proto-Oncogene Proteins / genetics
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Proto-Oncogene Proteins / physiology*
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RNA Interference*
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RNA, Small Interfering / administration & dosage
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RNA, Small Interfering / genetics*
Substances
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DNA-Binding Proteins
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Proto-Oncogene Proteins
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RNA, Small Interfering
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5-hydroxymethylcytosine
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5-Methylcytosine
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Cytosine
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Dioxygenases
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TET2 protein, human