A Novel Germline Variant in CSF3R Reduces N-Glycosylation and Exerts Potent Oncogenic Effects in Leukemia

Cancer Res. 2018 Dec 15;78(24):6762-6770. doi: 10.1158/0008-5472.CAN-18-1638. Epub 2018 Oct 22.

Abstract

: Mutations in the colony-stimulating factor 3 receptor (CSF3R) have been identified in the vast majority of patients with chronic neutrophilic leukemia and are present in other kinds of leukemia, such as acute myeloid leukemia. Here, we studied the function of novel germline variants in CSF3R at amino acid N610. These N610 substitutions were potently oncogenic and activated the receptor independently of its ligand GCSF. These mutations activated the JAK-STAT signaling pathway and conferred sensitivity to JAK inhibitors. Mass spectrometry revealed that the N610 residue is part of a consensus N-linked glycosylation motif in the receptor, usually linked to complex glycans. N610 was also the primary site of sialylation of the receptor. Membrane-proximal N-linked glycosylation was critical for maintaining the ligand dependence of the receptor. Mutation of the N610 site prevented membrane-proximal N-glycosylation of CSF3R, which then drove ligand-independent cellular expansion. Kinase inhibitors blocked growth of cells with an N610 mutation. This study expands the repertoire of oncogenic mutations in CSF3R that are therapeutically targetable and provides insight into the function of glycans in receptor regulation. SIGNIFICANCE: This study reveals the critical importance of membrane-proximal N-linked glycosylation of CSF3R for the maintenance of ligand dependency in leukemia.

Publication types

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

MeSH terms

  • Amino Acid Motifs
  • Animals
  • Binding Sites
  • Carcinogenesis*
  • Cell Membrane / metabolism
  • Disease Progression
  • Female
  • Gene Expression Regulation, Leukemic
  • Germ-Line Mutation*
  • Glycosylation
  • HEK293 Cells
  • Humans
  • Janus Kinases / metabolism
  • Leukemia / genetics*
  • Leukemia / metabolism
  • Leukemia, Myeloid, Acute / genetics
  • Leukemia, Neutrophilic, Chronic / genetics
  • Ligands
  • Mass Spectrometry
  • Mice
  • Mice, Inbred C57BL
  • Middle Aged
  • Mutation*
  • Proteomics
  • Receptors, Colony-Stimulating Factor / genetics*
  • Receptors, Colony-Stimulating Factor / metabolism
  • STAT Transcription Factors / metabolism
  • Sequence Analysis, DNA
  • Signal Transduction
  • Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization

Substances

  • CSF3R protein, human
  • Ligands
  • Receptors, Colony-Stimulating Factor
  • STAT Transcription Factors
  • Janus Kinases