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The Cell Type–Specific 5hmC Landscape and Dynamics of Healthy Human Hematopoiesis and TET2 -Mutant Preleukemia

Nakauchi, Yusuke, Azizi, Armon, Thomas, Daniel, Corces, M. Ryan, Reinisch, Andreas, Sharma, Rajiv, Cruz Hernandez, David, Köhnke, Thomas, Karigane, Daiki, Fan, Amy, Martinez-Krams, Daniel, Stafford, Melissa, Kaur, Satinder, Dutta, Ritika, Phan, Paul, Ediriwickrema, Asiri, McCarthy, Erin, Ning, Yuhong, Phillips, Tierney, Ellison, Christopher K., Guler, Gulfem D., Bergamaschi, Anna, Ku, Chin-Jen, Levy, Samuel, Majeti, Ravindra·2022·Blood Cancer Discovery
DOI 10.1158/2643-3230.bcd-21-0143

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Abstract

The conversion of 5-methylcytosine (5mC) to 5-hydroxymethylcytosine (5hmC) is a key step in DNA demethylation that is mediated by ten–eleven translocation (TET) enzymes, which require ascorbate/vitamin C. Here, we report the 5hmC landscape of normal hematopoiesis and identify cell type–specific 5hmC profiles associated with active transcription and chromatin accessibility of key hematopoietic regulators. We utilized CRISPR/Cas9 to model TET2 loss-of-function mutations in primary human hematopoietic stem and progenitor cells (HSPC). Disrupted cells exhibited increased colonies in serial replating, defective erythroid/megakaryocytic differentiation, and in vivo competitive advantage and myeloid skewing coupled with reduction of 5hmC at erythroid-associated gene loci. Azacitidine and ascorbate restored 5hmC abundance and slowed or reverted the expansion of TET2-mutant clones in vivo. These results demonstrate the key role of 5hmC in normal hematopoiesis and TET2-mutant phenotypes and raise the possibility of utilizing these agents to further our understanding of preleukemia and clonal hematopoiesis. Significance: We show that 5-hydroxymethylation profiles are cell type–specific and associated with transcriptional abundance and chromatin accessibility across human hematopoiesis. TET2 loss caused aberrant growth and differentiation phenotypes and disrupted 5hmC and transcriptional landscapes. Treatment of TET2 KO HSPCs with ascorbate or azacitidine reverted 5hmC profiles and restored aberrant phenotypes. This article is highlighted in the In This Issue feature, p. 265

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Cited by

Metadata

Authors
Nakauchi, Y., Azizi, A., Thomas, D., Corces, M. R., Reinisch, A., Sharma, R., Cruz Hernandez, D., Köhnke, T., Karigane, D., Fan, A., Martinez-Krams, D., Stafford, M., Kaur, S., Dutta, R., Phan, P., Ediriwickrema, A., McCarthy, E., Ning, Y., Phillips, T., Ellison, C. K., Guler, G. D., Bergamaschi, A., Ku, C.-J., Levy, S., & Majeti, R.
Affiliations
  • 1Department of Medicine, Division of Hematology, Stanford University School of Medicine, Stanford, California.
  • 11Bluestar Genomics Inc., San Mateo, California.
ORCID
Venue
Blood Cancer Discovery
Publisher
American Association for Cancer Research (AACR)
Published
2022-06-02
Volume
3
Issue
4
Pages
346-367
ISSN
  • 2643-3230
  • 2643-3249
DOI
10.1158/2643-3230.bcd-21-0143
Cited by
58
References
13
Language
en
Funders
NIH (1R01HL142637, 1R01CA251331)

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