Definition of a small core transcriptional circuit regulated by AML1-ETO

Mol Cell. 2021 Feb 4;81(3):530-545.e5. doi: 10.1016/j.molcel.2020.12.005. Epub 2020 Dec 30.

Abstract

Transcription factors regulate gene networks controlling normal hematopoiesis and are frequently deregulated in acute myeloid leukemia (AML). Critical to our understanding of the mechanism of cellular transformation by oncogenic transcription factors is the ability to define their direct gene targets. However, gene network cascades can change within minutes to hours, making it difficult to distinguish direct from secondary or compensatory transcriptional changes by traditional methodologies. To overcome this limitation, we devised cell models in which the AML1-ETO protein could be quickly degraded upon addition of a small molecule. The rapid kinetics of AML1-ETO removal, when combined with analysis of transcriptional output by nascent transcript analysis and genome-wide AML1-ETO binding by CUT&RUN, enabled the identification of direct gene targets that constitute a core AML1-ETO regulatory network. Moreover, derepression of this gene network was associated with RUNX1 DNA binding and triggered a transcription cascade ultimately resulting in myeloid differentiation.

Keywords: AML1-ETO; PRO-seq; PROTAC; RUNX1; RUNX1T1; degron tag; myeloid leukemia; nascent transcription.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Acetylation
  • Binding Sites
  • Binding, Competitive
  • Cell Differentiation
  • Cell Line, Tumor
  • Cell Proliferation
  • Cell Self Renewal
  • Core Binding Factor Alpha 2 Subunit / genetics
  • Core Binding Factor Alpha 2 Subunit / metabolism*
  • Fetal Blood / cytology
  • Gene Expression Regulation, Leukemic
  • Gene Regulatory Networks
  • HEK293 Cells
  • Hematopoietic Stem Cells / metabolism*
  • Hematopoietic Stem Cells / pathology
  • Histones / metabolism
  • Humans
  • Leukemia, Myeloid, Acute / genetics
  • Leukemia, Myeloid, Acute / metabolism*
  • Leukemia, Myeloid, Acute / pathology
  • Neoplastic Stem Cells / metabolism*
  • Neoplastic Stem Cells / pathology
  • Oncogene Proteins, Fusion / genetics
  • Oncogene Proteins, Fusion / metabolism*
  • Protein Binding
  • Proteolysis
  • Proto-Oncogene Proteins / genetics
  • Proto-Oncogene Proteins / metabolism
  • RNA, Neoplasm / biosynthesis*
  • RNA, Neoplasm / genetics
  • RUNX1 Translocation Partner 1 Protein / genetics
  • RUNX1 Translocation Partner 1 Protein / metabolism*
  • Repressor Proteins / genetics
  • Repressor Proteins / metabolism
  • Time Factors
  • Transcription, Genetic*
  • Transcriptome

Substances

  • AML1-ETO fusion protein, human
  • Core Binding Factor Alpha 2 Subunit
  • GFI1B protein, human
  • Histones
  • Oncogene Proteins, Fusion
  • Proto-Oncogene Proteins
  • RNA, Neoplasm
  • RUNX1 Translocation Partner 1 Protein
  • RUNX1 protein, human
  • Repressor Proteins