High levels of soluble CD73 unveil resistance to BRAF inhibitors in melanoma cells

Biomed Pharmacother. 2024 Aug:177:117033. doi: 10.1016/j.biopha.2024.117033. Epub 2024 Jun 27.

Abstract

Melanoma cells express high levels of CD73 that produce extracellular immunosuppressive adenosine. Changes in the CD73 expression occur in response to tumor environmental factors, contributing to tumor phenotype plasticity and therapeutic resistance. Previously, we have observed that CD73 expression can be up-regulated on the surface of melanoma cells in response to nutritional stress. Here, we explore the mechanism by which melanoma cells release soluble CD73 under low nutrient availability and whether this might be affected by agents targeting the proto-oncogene B-Raf (BRAF). We found that starved melanoma cells can release high levels of CD73, able to convert AMP into adenosine, and this activity is abrogated by selective CD73 inhibitors, APCP or PSB-12489. The release of CD73 from melanoma cells is mediated by the matrix metalloproteinase MMP-9. Indeed, MMP-9 inhibitors significantly reduce the levels of CD73 released from the cells, while its surface levels increase. Of relevance, melanoma cells, harboring an activating BRAF mutation, upon treatment with dabrafenib or vemurafenib, show a strong reduction of CD73 cell expression and reduced levels of CD73 released into the extracellular space. Conversely, melanoma cells resistant to dabrafenib show high expression of membrane-bound CD73 and soluble CD73 released into the culture medium. In summary, our data indicate that CD73 is released from melanoma cells. The expression of CD73 is associated with response to BRAF inhibitors. Melanoma cells developing resistance to dabrafenib show increased expression of CD73, including soluble CD73 released from cells, suggesting that CD73 is involved in acquiring resistance to treatment.

Keywords: BRAF inhibitor; CD73; MMP9; melanoma cells.

MeSH terms

  • 5'-Nucleotidase* / genetics
  • 5'-Nucleotidase* / metabolism
  • Cell Line, Tumor
  • Drug Resistance, Neoplasm* / drug effects
  • GPI-Linked Proteins / genetics
  • GPI-Linked Proteins / metabolism
  • Humans
  • Imidazoles / pharmacology
  • Matrix Metalloproteinase 9 / metabolism
  • Melanoma* / drug therapy
  • Melanoma* / genetics
  • Melanoma* / metabolism
  • Melanoma* / pathology
  • Oximes / pharmacology
  • Protein Kinase Inhibitors / pharmacology
  • Proto-Oncogene Mas*
  • Proto-Oncogene Proteins B-raf* / antagonists & inhibitors
  • Proto-Oncogene Proteins B-raf* / genetics
  • Proto-Oncogene Proteins B-raf* / metabolism
  • Vemurafenib* / pharmacology

Substances

  • Proto-Oncogene Proteins B-raf
  • 5'-Nucleotidase
  • Proto-Oncogene Mas
  • MAS1 protein, human
  • NT5E protein, human
  • BRAF protein, human
  • Vemurafenib
  • Oximes
  • Matrix Metalloproteinase 9
  • Imidazoles
  • dabrafenib
  • Protein Kinase Inhibitors
  • GPI-Linked Proteins