Leukocytes as a reservoir of circulating oncogenic DNA and regulatory targets of tumor-derived extracellular vesicles

J Thromb Haemost. 2018 Sep;16(9):1800-1813. doi: 10.1111/jth.14222. Epub 2018 Aug 16.

Abstract

Essentials Tumor-bearing mice were employed to follow oncogenic HRAS sequences in plasma, and blood cells. Cancer DNA accumulated in leukocytes above levels detected in exosomes, platelets and plasma. Extracellular vesicles and nucleosomes are required for uptake of tumor DNA by leukocytes. Uptake of tumor-derived extracellular vesicles by leukocytes triggers coagulant phenotype.

Summary: Background Tumor-derived extracellular vesicles (EVs) and free nucleosomes (NSs) carry into the circulation a wealth of cancer-specific, bioactive and poorly understood molecular cargoes, including genomic DNA (gDNA). Objective Here we investigated the distribution of extracellular oncogenic gDNA sequences (HRAS and HER2) in the circulation of tumor-bearing mice. Methods and Results Surprisingly, circulating leukocytes (WBCs), especially neutrophils, contained the highest levels of mutant gDNA, which exceeded the amount of this material recovered from soluble fractions of plasma, circulating EVs, platelets, red blood cells (RBCs) and peripheral organs, as quantified by digital droplet PCR (ddPCR). Tumor excision resulted in disappearance of the WBC-associated gDNA signal within 2-9 days, which is in line with the expected half-life of these cells. EVs and nucleosomes were essential for the uptake of tumor-derived extracellular DNA by neutrophil-like cells and impacted their phenotype. Indeed, the exposure of granulocytic HL-60 cells to EVs from HRAS-driven cancer cells resulted in a selective increase in tissue factor (TF) procoagulant activity and interleukin 8 (IL-8) production. The levels of circulating thrombin-antithrombin complexes (TAT) were markedly elevated in mice harboring HRAS-driven xenografts. Conclusions Myeloid cells may represent a hitherto unrecognized reservoir of cancer-derived, EV/NS-associated oncogenic gDNA in the circulation, and a possible novel platform for liquid biopsy in cancer. In addition, uptake of this material alters the phenotype of myeloid cells, induces procoagulant and proinflammatory activity and may contribute to systemic effects associated with cancer.

Keywords: extracellular vesicles; neutrophils; nucleosomes; oncogenes; thrombosis.

Publication types

  • Comparative Study
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antithrombin III
  • Blood Platelets / chemistry
  • Breast Neoplasms / pathology
  • Cell Line, Tumor
  • Cell Survival
  • Cell Transformation, Neoplastic
  • DNA, Neoplasm / blood*
  • DNA, Neoplasm / pharmacokinetics
  • Exosomes / chemistry
  • Extracellular Vesicles / chemistry*
  • Female
  • Genes, erbB-2*
  • Genes, ras*
  • HL-60 Cells
  • Heterografts
  • Humans
  • Interleukin-8 / biosynthesis
  • Mice
  • Mice, SCID
  • Myeloid Cells / chemistry*
  • Myeloid Cells / metabolism
  • Neoplasm Transplantation
  • Neutrophils / chemistry*
  • Neutrophils / metabolism
  • Nucleosomes / chemistry
  • Peptide Hydrolases / blood
  • Plasma / chemistry
  • Rats
  • THP-1 Cells
  • Thromboplastin / biosynthesis
  • Tumor Burden

Substances

  • DNA, Neoplasm
  • Interleukin-8
  • Nucleosomes
  • antithrombin III-protease complex
  • Antithrombin III
  • Thromboplastin
  • Peptide Hydrolases

Grant support