GDF6-CD99 Signaling Regulates Src and Ewing Sarcoma Growth

Cell Rep. 2020 Nov 3;33(5):108332. doi: 10.1016/j.celrep.2020.108332.

Abstract

We report here that the autocrine signaling mediated by growth and differentiation factor 6 (GDF6), a member of the bone morphogenetic protein (BMP) family of cytokines, maintains Ewing sarcoma growth by preventing Src hyperactivation. Surprisingly, Ewing sarcoma depends on the prodomain, not the BMP domain, of GDF6. We demonstrate that the GDF6 prodomain is a ligand for CD99, a transmembrane protein that has been widely used as a marker of Ewing sarcoma. The binding of the GDF6 prodomain to the CD99 extracellular domain results in recruitment of CSK (C-terminal Src kinase) to the YQKKK motif in the intracellular domain of CD99, inhibiting Src activity. GDF6 silencing causes hyperactivation of Src and p21-dependent growth arrest. We demonstrate that two GDF6 prodomain mutants linked to Klippel-Feil syndrome are hyperactive in CD99-Src signaling. These results reveal a cytokine signaling pathway that regulates the CSK-Src axis and cancer cell proliferation and suggest the gain-of-function activity for disease-causing GDF6 mutants.

Keywords: CD99; CSK; Ewing sarcoma; GDF6; Klippel-Feil syndrome; Src; proteomics; secretome.

Publication types

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

MeSH terms

  • 12E7 Antigen / metabolism*
  • Animals
  • CSK Tyrosine-Protein Kinase / metabolism
  • Cell Proliferation
  • Down-Regulation
  • Gene Expression Regulation, Neoplastic
  • Growth Differentiation Factor 6 / chemistry
  • Growth Differentiation Factor 6 / metabolism*
  • Humans
  • Klippel-Feil Syndrome / genetics
  • Mice, SCID
  • Mutation / genetics
  • Oncogene Proteins, Fusion / metabolism
  • Protein Domains
  • Proteome / metabolism
  • Proteomics
  • Proto-Oncogene Protein c-fli-1 / metabolism
  • RNA-Binding Protein EWS / metabolism
  • Sarcoma, Ewing / metabolism*
  • Sarcoma, Ewing / pathology*
  • Signal Transduction*
  • Transcription, Genetic
  • src-Family Kinases / metabolism*

Substances

  • 12E7 Antigen
  • CD99 protein, human
  • EWS-FLI fusion protein
  • GDF6 protein, human
  • Growth Differentiation Factor 6
  • Oncogene Proteins, Fusion
  • Proteome
  • Proto-Oncogene Protein c-fli-1
  • RNA-Binding Protein EWS
  • CSK Tyrosine-Protein Kinase
  • src-Family Kinases
  • CSK protein, human