Conditional knockin of Dnmt3a R878H initiates acute myeloid leukemia with mTOR pathway involvement

Proc Natl Acad Sci U S A. 2017 May 16;114(20):5237-5242. doi: 10.1073/pnas.1703476114. Epub 2017 May 1.

Abstract

DNMT3A is frequently mutated in acute myeloid leukemia (AML). To explore the features of human AML with the hotspot DNMT3A R882H mutation, we generated Dnmt3a R878H conditional knockin mice, which developed AML with enlarged Lin-Sca1+cKit+ cell compartments. The transcriptome and DNA methylation profiling of bulk leukemic cells and the single-cell RNA sequencing of leukemic stem/progenitor cells revealed significant changes in gene expression and epigenetic regulatory patterns that cause differentiation arrest and growth advantage. Consistent with leukemic cell accumulation in G2/M phase, CDK1 was up-regulated due to mTOR activation associated with DNA hypomethylation. Overexpressed CDK1-mediated EZH2 phosphorylation resulted in an abnormal trimethylation of H3K27 profile. The mTOR inhibitor rapamycin elicited a significant therapeutic response in Dnmt3aR878H/WT mice.

Keywords: Dnmt3a R878H mutation; conditional knockin mice; leukemia; mTOR inhibitor; single-cell RNA-seq.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Base Sequence
  • Cell Differentiation
  • DNA (Cytosine-5-)-Methyltransferases / genetics*
  • DNA (Cytosine-5-)-Methyltransferases / metabolism*
  • DNA Methylation
  • DNA Methyltransferase 3A
  • DNA Modification Methylases / metabolism
  • Disease Models, Animal
  • Gene Expression Profiling / methods
  • Gene Knock-In Techniques / methods
  • Leukemia, Myeloid, Acute / genetics*
  • Leukemia, Myeloid, Acute / metabolism
  • Mice
  • Mutation
  • TOR Serine-Threonine Kinases / metabolism
  • Transcriptome

Substances

  • DNMT3A protein, human
  • Dnmt3a protein, mouse
  • DNA Modification Methylases
  • DNA (Cytosine-5-)-Methyltransferases
  • DNA Methyltransferase 3A
  • mTOR protein, mouse
  • TOR Serine-Threonine Kinases