Hematopoietic stem cells (HSCs) regenerate the peripheral blood throughout life via their capacity to both self-renew and differentiate. A critica role for DNA methyltransferases in maintaining effective HSC function has recently come to light through a series of studies in mouse and man. Our laboratory recently found that that DNA methyltransferase 3a (DNMT3A) was highly expressed in murine HSCs, and its conditional loss promoted dramatic self-renewal at the expense of differentiation. In humans, DNMT3A was found to be mutated in around 10% of myelodysplastic syndrome (MDS) patients, and around 20% of patients with either myeloid or lymphoid leukemia. DNMT3A mutations appear to arise in the HSCs, conferring an advantage to mutant cells, as these mutations are found in non-malignant as well as malignant clones, and HSC progeny of all lineages. Finally, cells bearing DNMT3A mutations are common in the peripheral blood of individuals over the age of 50 who exhibit a clonal dominance in their peripheral blood. Together, these studies from both mouse and humans incontrovertibly point to a central role for DNMT3A in maintaining normal hematopoiesis. Nevertheless, how these mutations confer an advantage to HSCs and contribute to clonal expansion as well as to the development of MDS and leukemia remains unclear. Gaining insight into these mechanisms is the overarching goal of this project. Here, we will determine whether DNMT3a mutations lead to blocked differentiation, whether DNA methylation function per se is critical to the hematopoietic impact, and whether different DNMT3A mutations have similar roles. Specifically, we will examine the role in differentiation by mapping DNA methylation changes throughout HSC differentiation, determining the localization of DNMT3A protein throughout the genome, modulating genes involved in self-renewal or differentiation in DNMT3a-KO HSCs, and by modulating DNA methylation. In addition, we will determine whether DNA methylation is required for DNMT3A to exert its role in HSCs. We will add back to KO HSCs specific DNMTs that retain or lack catalysis function, and examining the progeny for DNA methylation and HSC activity. Finally, we will examine commonly occurring DNMT3A mutants such as the R882 form to determine their impact on DNA methylation and hematopoiesis. Together, these studies will definitively link differentiation inhibition with DNMT3A loss-of-function and reveal key targets that could be modulated for therapeutic strategies. These studies will also demonstrate whether catalytic DNA methyltransferase activity is required for full HSC function, and lend insights into the impact of some of common DNMT3A mutations. Our lab is uniquely placed to contribute to this growing area given our pioneering work on the role of DNMT3A in HSCs and our generation of several novel tools and reagents to pursue the proposed questions.
Blood is regenerated throughout life by specialized stem cells. Normal function of these stem cells requires action of a DNA modifying enzyme called DNMT3A, and it mutation is associated with abnormal blood production as well myelodysplastic syndrome (MDS) and leukemia development. The overall goal of this project is to understand what aspects of blood development require this enzyme and how it functions, with a view to being able to modulate these properties for potential therapeutic benefit.
| Jeong, Mira; Park, Hyun Jung; Celik, Hamza et al. (2018) Loss of Dnmt3a Immortalizes Hematopoietic Stem Cells In Vivo. Cell Rep 23:1-10 |
| Hsu, Joanne I; Dayaram, Tajhal; Tovy, Ayala et al. (2018) PPM1D Mutations Drive Clonal Hematopoiesis in Response to Cytotoxic Chemotherapy. Cell Stem Cell 23:700-713.e6 |
| Brunetti, Lorenzo; Gundry, Michael C; Sorcini, Daniele et al. (2018) Mutant NPM1 Maintains the Leukemic State through HOX Expression. Cancer Cell 34:499-512.e9 |
| Su, Jianzhong; Huang, Yung-Hsin; Cui, Xiaodong et al. (2018) Homeobox oncogene activation by pan-cancer DNA hypermethylation. Genome Biol 19:108 |
| Tan, Qiumin; Brunetti, Lorenzo; Rousseaux, Maxime W C et al. (2018) Loss of Capicua alters early T cell development and predisposes mice to T cell lymphoblastic leukemia/lymphoma. Proc Natl Acad Sci U S A 115:E1511-E1519 |
| Jeong, Mira; Guzman, Anna G; Goodell, Margaret A (2017) Genome-Wide Analysis of DNA Methylation in Hematopoietic Cells: DNA Methylation Analysis by WGBS. Methods Mol Biol 1633:137-149 |
| Huang, Yung-Hsin; Su, Jianzhong; Lei, Yong et al. (2017) DNA epigenome editing using CRISPR-Cas SunTag-directed DNMT3A. Genome Biol 18:176 |
| Lei, Yong; Zhang, Xiaotian; Su, Jianzhong et al. (2017) Targeted DNA methylation in vivo using an engineered dCas9-MQ1 fusion protein. Nat Commun 8:16026 |
| Brunetti, Lorenzo; Gundry, Michael C; Goodell, Margaret A (2017) DNMT3A in Leukemia. Cold Spring Harb Perspect Med 7: |
| Rau, Rachel E; Rodriguez, Benjamin A; Luo, Min et al. (2016) DOT1L as a therapeutic target for the treatment of DNMT3A-mutant acute myeloid leukemia. Blood 128:971-81 |
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