HyperTRIBE uncovers increased MUSASHI-2 RNA binding activity and differential regulation in leukemic stem cells

Nat Commun. 2020 Apr 24;11(1):2026. doi: 10.1038/s41467-020-15814-8.

Abstract

The cell-context dependency for RNA binding proteins (RBPs) mediated control of stem cell fate remains to be defined. Here we adapt the HyperTRIBE method using an RBP fused to a Drosophila RNA editing enzyme (ADAR) to globally map the mRNA targets of the RBP MSI2 in mammalian adult normal and malignant stem cells. We reveal a unique MUSASHI-2 (MSI2) mRNA binding network in hematopoietic stem cells that changes during transition to multipotent progenitors. Additionally, we discover a significant increase in RNA binding activity of MSI2 in leukemic stem cells compared with normal hematopoietic stem and progenitor cells, resulting in selective regulation of MSI2's oncogenic targets. This provides a basis for MSI2 increased dependency in leukemia cells compared to normal cells. Moreover, our study provides a way to measure RBP function in rare cells and suggests that RBPs can achieve differential binding activity during cell state transition independent of gene expression.

Publication types

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

MeSH terms

  • Adenosine Deaminase / genetics
  • Animals
  • Binding Sites / genetics
  • Cell Differentiation / genetics*
  • Disease Models, Animal
  • Drosophila Proteins / genetics
  • Gene Expression Regulation, Leukemic
  • Gene Regulatory Networks
  • HEK293 Cells
  • Hematopoietic Stem Cells / pathology*
  • Humans
  • Leukemia / blood
  • Leukemia / genetics*
  • Leukemia / pathology
  • Mice
  • Mice, Knockout
  • Neoplastic Stem Cells / pathology*
  • Primary Cell Culture
  • RNA, Messenger / metabolism
  • RNA-Binding Proteins / genetics
  • RNA-Binding Proteins / metabolism*
  • RNA-Seq
  • Recombinant Fusion Proteins / genetics
  • Recombinant Fusion Proteins / metabolism

Substances

  • Drosophila Proteins
  • MSI2 protein, human
  • Msi2h protein, mouse
  • RNA, Messenger
  • RNA-Binding Proteins
  • Recombinant Fusion Proteins
  • Adar protein, Drosophila
  • Adenosine Deaminase