Nascent transcript and single-cell RNA-seq analysis defines the mechanism of action of the LSD1 inhibitor INCB059872 in myeloid leukemia

Gene. 2020 Aug 20:752:144758. doi: 10.1016/j.gene.2020.144758. Epub 2020 May 15.

Abstract

Drugs targeting chromatin-modifying enzymes have entered clinical trials for myeloid malignancies, including INCB059872, a selective irreversible inhibitor of Lysine-Specific Demethylase 1 (LSD1). While initial studies of LSD1 inhibitors suggested these compounds may be used to induce differentiation of acute myeloid leukemia (AML), the mechanisms underlying this effect and dose-limiting toxicities are not well understood. Here, we used precision nuclear run-on sequencing (PRO-seq) and ChIP-seq in AML cell lines to probe for the earliest regulatory events associated with INCB059872 treatment. The changes in nascent transcription could be traced back to a loss of CoREST activity and activation of GFI1-regulated genes. INCB059872 is in phase I clinical trials, and we evaluated a pre-treatment bone marrow sample of a patient who showed a clinical response to INCB059872 while being treated with azacitidine. We used single-cell RNA-sequencing (scRNA-seq) to show that INCB059872 caused a shift in gene expression that was again associated with GFI1/GFI1B regulation. Finally, we treated mice with INCB059872 and performed scRNA-seq of lineage-negative bone marrow cells, which showed that INCB059872 triggered accumulation of megakaryocyte early progenitor cells with gene expression hallmarks of stem cells. Accumulation of these stem/progenitor cells may contribute to the thrombocytopenia observed in patients treated with LSD1 inhibitors.

Keywords: CoREST; GFI1; GFI1B; Histone acetylation; Histone demethylation; KDM1A; LSD1; Myeloid leukemia.

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Cell Differentiation / drug effects
  • Cell Line, Tumor
  • DNA-Binding Proteins / genetics
  • DNA-Binding Proteins / metabolism
  • Exome Sequencing / methods
  • Female
  • Gene Expression Regulation, Leukemic / drug effects*
  • Histone Demethylases / antagonists & inhibitors*
  • Histone Demethylases / genetics
  • Histone Demethylases / metabolism
  • Humans
  • Leukemia, Myeloid, Acute / genetics
  • Leukemia, Myeloid, Acute / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • RNA-Seq
  • Single-Cell Analysis / methods
  • Stem Cells / metabolism
  • THP-1 Cells
  • Transcription Factors / genetics
  • Transcription Factors / metabolism

Substances

  • Antineoplastic Agents
  • DNA-Binding Proteins
  • GFI1 protein, human
  • Transcription Factors
  • Histone Demethylases
  • KDM1A protein, human