Detection and targeting of splicing deregulation in pediatric acute myeloid leukemia stem cells

Cell Rep Med. 2023 Mar 21;4(3):100962. doi: 10.1016/j.xcrm.2023.100962. Epub 2023 Mar 7.

Abstract

Pediatric acute myeloid leukemia (pAML) is typified by high relapse rates and a relative paucity of somatic DNA mutations. Although seminal studies show that splicing factor mutations and mis-splicing fuel therapy-resistant leukemia stem cell (LSC) generation in adults, splicing deregulation has not been extensively studied in pAML. Herein, we describe single-cell proteogenomics analyses, transcriptome-wide analyses of FACS-purified hematopoietic stem and progenitor cells followed by differential splicing analyses, dual-fluorescence lentiviral splicing reporter assays, and the potential of a selective splicing modulator, Rebecsinib, in pAML. Using these methods, we discover transcriptomic splicing deregulation typified by differential exon usage. In addition, we discover downregulation of splicing regulator RBFOX2 and CD47 splice isoform upregulation. Importantly, splicing deregulation in pAML induces a therapeutic vulnerability to Rebecsinib in survival, self-renewal, and lentiviral splicing reporter assays. Taken together, the detection and targeting of splicing deregulation represent a potentially clinically tractable strategy for pAML therapy.

Keywords: CD47; RBFOX2; SF3B1; embryonic stem cells; hematopoietic stem cells; pediatric AML; pre-mRNA splicing; splicing modulation.

Publication types

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

MeSH terms

  • Adult
  • Child
  • Humans
  • Leukemia, Myeloid, Acute* / drug therapy
  • Leukemia, Myeloid, Acute* / genetics
  • Mutation
  • Protein Isoforms / genetics
  • RNA Splicing / genetics
  • RNA Splicing Factors / genetics
  • Repressor Proteins / genetics
  • Stem Cells*

Substances

  • Protein Isoforms
  • RBFOX2 protein, human
  • RNA Splicing Factors
  • Repressor Proteins