title: DOT1L as a therapeutic target for the treatment of DNMT3A-mutant acute myeloid leukemia.
abstract: Mutations in DNA methyltransferase 3A (DNMT3A) are common in acute myeloid leukemia and portend a poor prognosis; thus, new therapeutic strategies are needed. The likely mechanism by which DNMT3A loss contributes to leukemogenesis is altered DNA methylation and the attendant gene expression changes; however, our current understanding is incomplete. We observed that murine hematopoietic stem cells (HSCs) in which Dnmt3a had been conditionally deleted markedly overexpress the histone 3 lysine 79 (H3K79) methyltransferase, Dot1l. We demonstrate that Dnmt3a(-/-) HSCs have increased H3K79 methylation relative to wild-type (WT) HSCs, with the greatest increases noted at DNA methylation canyons, which are regions highly enriched for genes dysregulated in leukemia and prone to DNA methylation loss with Dnmt3a deletion. These findings led us to explore DOT1L as a therapeutic target for the treatment of DNMT3A-mutant AML. We show that pharmacologic inhibition of DOT1L resulted in decreased expression of oncogenic canyon-associated genes and led to dose- and time-dependent inhibition of proliferation, induction of apoptosis, cell-cycle arrest, and terminal differentiation in DNMT3A-mutant cell lines in vitro. We show in vivo efficacy of the DOT1L inhibitor EPZ5676 in a nude rat xenograft model of DNMT3A-mutant AML. DOT1L inhibition was also effective against primary patient DNMT3A-mutant AML samples, reducing colony-forming capacity (CFC) and inducing terminal differentiation in vitro. These studies suggest that DOT1L may play a critical role in DNMT3A-mutant leukemia. With pharmacologic inhibitors of DOT1L already in clinical trials, DOT1L could be an immediately actionable therapeutic target for the treatment of this poor prognosis disease.
authors.affiliation: Department of Pediatrics, Baylor College of Medicine and Texas Children's Hospital, Houston, TX; Dan L. Duncan Cancer Center.
authors.forename: Rachel E
authors.initials: RE
authors.lastname: Rau
authors.affiliation: Dan L. Duncan Cancer Center, Department of Molecular and Cellular Biology.
authors.forename: Benjamin A
authors.initials: BA
authors.lastname: Rodriguez
authors.affiliation: Department of Molecular and Human Genetics, Stem Cells and Regenerative Medicine Center, and Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX;
authors.forename: Min
authors.initials: M
authors.lastname: Luo
authors.affiliation: Department of Molecular and Human Genetics, Stem Cells and Regenerative Medicine Center, and Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX;
authors.forename: Mira
authors.initials: M
authors.lastname: Jeong
authors.affiliation: Department of Molecular and Human Genetics, Stem Cells and Regenerative Medicine Center, and Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX;
authors.forename: Allison
authors.initials: A
authors.lastname: Rosen
authors.affiliation: Department of Pediatrics, Baylor College of Medicine and Texas Children's Hospital, Houston, TX;
authors.forename: Jason H
authors.initials: JH
authors.lastname: Rogers
authors.affiliation: Epizyme, Inc., Cambridge, MA;
authors.forename: Carly T
authors.initials: CT
authors.lastname: Campbell
authors.affiliation: Epizyme, Inc., Cambridge, MA;
authors.forename: Scott R
authors.initials: SR
authors.lastname: Daigle
authors.affiliation: Department of Pharmacology, Baylor College of Medicine, Houston, TX;
authors.forename: Lishing
authors.initials: L
authors.lastname: Deng
authors.affiliation: Department of Pharmacology, Baylor College of Medicine, Houston, TX;
authors.forename: Yongcheng
authors.initials: Y
authors.lastname: Song
authors.affiliation: Genome Damage and Stability Centre, and.
authors.forename: Steve
authors.initials: S
authors.lastname: Sweet
authors.affiliation: Department of Haematology, Brighton and Sussex Medical School, University of Sussex, Brighton, United Kingdom; and.
authors.forename: Timothy
authors.initials: T
authors.lastname: Chevassut
authors.affiliation: Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX.
authors.forename: Michael
authors.identifier: 0000-0002-1144-1958
authors.initials: M
authors.lastname: Andreeff
authors.affiliation: Department of Leukemia, The University of Texas MD Anderson Cancer Center, Houston, TX.
authors.forename: Steven M
authors.initials: SM
authors.lastname: Kornblau
authors.affiliation: Dan L. Duncan Cancer Center, Department of Molecular and Cellular Biology.
authors.forename: Wei
authors.initials: W
authors.lastname: Li
authors.affiliation: Department of Pediatrics, Baylor College of Medicine and Texas Children's Hospital, Houston, TX; Dan L. Duncan Cancer Center, Department of Molecular and Human Genetics, Stem Cells and Regenerative Medicine Center, and Center for Cell and Gene Therapy, Baylor College of Medicine, Houston, TX;
authors.forename: Margaret A
authors.initials: MA
authors.lastname: Goodell
mesh_terms: D000241:Adenosine / Q000031:analogs & derivatives / Q000494:pharmacology / Q000627:therapeutic use; D000818:Animals; D017209:Apoptosis; D059447:Cell Cycle Checkpoints; D002454:Cell Differentiation; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D004248:DNA (Cytosine-5-)-Methyltransferases / Q000235:genetics*; D000090205:DNA Methyltransferase 3A; D015973:Gene Expression Regulation, Leukemic / Q000187:drug effects; D006412:Hematopoietic Stem Cells / Q000187:drug effects / Q000378:metabolism; D011495:Histone-Lysine N-Methyltransferase; D006657:Histones / Q000378:metabolism; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000188:drug therapy* / Q000201:enzymology* / Q000235:genetics / Q000473:pathology; D008239:Lysine / Q000378:metabolism; D008745:Methylation; D008780:Methyltransferases / Q000235:genetics* / Q000378:metabolism*; D008810:Mice, Inbred C57BL; D058990:Molecular Targeted Therapy; D009154:Mutation / Q000235:genetics*; D010671:Phenylurea Compounds / Q000494:pharmacology / Q000627:therapeutic use; D051381:Rats; D013997:Time Factors; D014410:Tumor Stem Cell Assay; D023041:Xenograft Model Antitumor Assays
chemical_list: C000717967:DNMT3A protein, human; C000715535:Dnmt3a protein, mouse; C581325:EPZ004777; D006657:Histones; D010671:Phenylurea Compounds; C473477:DOT1L protein, human; C481527:Dot1l protein, mouse; D008780:Methyltransferases; D004248:DNA (Cytosine-5-)-Methyltransferases; D000090205:DNA Methyltransferase 3A; D011495:Histone-Lysine N-Methyltransferase; D008239:Lysine; D000241:Adenosine