Tracing Cellular Origin of Human Exosomes Using Multiplex Proximity Extension Assays

Mol Cell Proteomics. 2017 Mar;16(3):502-511. doi: 10.1074/mcp.M116.064725. Epub 2017 Jan 22.

Abstract

Extracellular vesicles (EVs) are membrane-coated objects such as exosomes and microvesicles, released by many cell-types. Their presence in body fluids and the variable surface composition and content render them attractive potential biomarkers. The ability to determine their cellular origin could greatly move the field forward. We used multiplex proximity extension assays (PEA) to identify with high specificity and sensitivity the protein profiles of exosomes of different origins, including seven cell lines and two different body fluids. By comparing cells and exosomes, we successfully identified the cells originating the exosomes. Furthermore, by principal component analysis of protein patterns human milk EVs and prostasomes released from prostate acinar cells clustered with cell lines from breast and prostate tissues, respectively. Milk exosomes uniquely expressed CXCL5, MIA, and KLK6, whereas prostasomes carried NKX31, GSTP1, and SRC, highlighting that EVs originating from different origins express distinct proteins. In conclusion, PEA provides a powerful protein screening tool in exosome research, for purposes of identifying the cell source of exosomes, or new biomarkers in diseases such as cancer and inflammation.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Biomarkers / metabolism*
  • Body Fluids / metabolism*
  • Cell Line
  • Cell-Derived Microparticles / metabolism*
  • Exosomes / metabolism*
  • Female
  • High-Throughput Screening Assays / methods*
  • Humans
  • K562 Cells
  • MCF-7 Cells
  • Male
  • Milk, Human / metabolism
  • Organ Specificity
  • Principal Component Analysis
  • Prostate / metabolism

Substances

  • Biomarkers