Defining the epigenetic status of blood cells using a cyanine-based fluorescent probe for PRMT1

Blood Adv. 2018 Nov 13;2(21):2829-2836. doi: 10.1182/bloodadvances.2018020727.

Abstract

Dynamic regulation of histone modification enzymes such as PRMT1 (protein arginine methyltransferase 1) determines the ordered epigenetic transitions in hematopoiesis. Sorting cells according to the expression levels of histone modification enzymes may further define subpopulations in hematopoietic lineages with unique differentiation potentials that are presently defined by surface markers. We discovered a vital near infrared dye, E84, that fluoresces brightly following binding to PRMT1 and excitation with a red laser. The staining intensity as measured by flow cytometry is correlated with the PRMT1 expression level. Importantly, E84 staining has no apparent negative effect on the proliferation of the labeled cells. Given that long-term hematopoietic stem cells (LT-HSCs) produce low levels of PRMT1, we used E84 to sort LT-HSCs from mouse bone marrow. We found that SLAM (the signalling lymphocyte activation molecule family) marker-positive LT-HSCs were enriched in the E84low cell fraction. We then performed bone marrow transplantations with E84high or E84low Lin-Sca1+Kit+ (LSK) cells and showed that whole blood cell lineages were successfully reconstituted 16 weeks after transplanting 200 E84low LSK cells. Thus, E84 is a useful new tool to probe the role of PRMT1 in hematopoiesis and leukemogenesis. Developing E84 and other small molecules to label histone modification enzymes provides a convenient approach without modifying gene loci to study the interaction between hematopoietic stem/progenitor cell epigenetic status and differentiation state.

Publication types

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

MeSH terms

  • Animals
  • Ataxin-1 / metabolism
  • Blood Cells / metabolism*
  • Blood Cells / pathology
  • Bone Marrow Cells / metabolism
  • Bone Marrow Cells / pathology
  • Bone Marrow Transplantation
  • Carbocyanines / chemistry*
  • Cell Lineage
  • Epigenesis, Genetic*
  • Flow Cytometry / methods
  • Fluorescent Dyes / chemistry*
  • HEK293 Cells
  • Hematopoietic Stem Cells / metabolism
  • Hematopoietic Stem Cells / pathology
  • Humans
  • Leukocyte Common Antigens / metabolism
  • Mice
  • Protein-Arginine N-Methyltransferases / genetics*
  • Protein-Arginine N-Methyltransferases / metabolism
  • Proto-Oncogene Proteins c-kit / metabolism

Substances

  • Ataxin-1
  • Carbocyanines
  • Fluorescent Dyes
  • Protein-Arginine N-Methyltransferases
  • Proto-Oncogene Proteins c-kit
  • Leukocyte Common Antigens