Enriched protein screening of human bone marrow mesenchymal stromal cell secretions reveals MFAP5 and PENK as novel IL-10 modulators

Mol Ther. 2014 May;22(5):999-1007. doi: 10.1038/mt.2014.17. Epub 2014 Feb 5.

Abstract

The secreted proteins from a cell constitute a natural biologic library that can offer significant insight into human health and disease. Discovering new secreted proteins from cells is bounded by the limitations of traditional separation and detection tools to physically fractionate and analyze samples. Here, we present a new method to systematically identify bioactive cell-secreted proteins that circumvent traditional proteomic methods by first enriching for protein candidates by differential gene expression profiling. The bone marrow stromal cell secretome was analyzed using enriched gene expression datasets in combination with potency assay testing. Four proteins expressed by stromal cells with previously unknown anti-inflammatory properties were identified, two of which provided a significant survival benefit to mice challenged with lethal endotoxic shock. Greater than 85% of secreted factors were recaptured that were otherwise undetected by proteomic methods, and remarkable hit rates of 18% in vitro and 9% in vivo were achieved.

Publication types

  • Research Support, N.I.H., Extramural

MeSH terms

  • Animals
  • Bone Marrow Cells / immunology
  • Bone Marrow Cells / metabolism
  • Contractile Proteins / genetics*
  • Contractile Proteins / metabolism*
  • Enkephalins / genetics*
  • Enkephalins / metabolism
  • Extracellular Matrix Proteins / genetics
  • Extracellular Matrix Proteins / metabolism*
  • Gene Expression Profiling
  • Glycoproteins / genetics*
  • Humans
  • Intercellular Signaling Peptides and Proteins
  • Interleukin-10 / genetics
  • Interleukin-10 / metabolism*
  • Mesenchymal Stem Cells / immunology
  • Mesenchymal Stem Cells / metabolism
  • Mice
  • Protein Biosynthesis / genetics
  • Protein Precursors / genetics*
  • Protein Precursors / metabolism
  • Proteins / genetics
  • Proteins / metabolism*
  • Proteomics
  • Shock, Septic / genetics
  • Shock, Septic / therapy*

Substances

  • Contractile Proteins
  • Enkephalins
  • Extracellular Matrix Proteins
  • Glycoproteins
  • IL10 protein, human
  • Intercellular Signaling Peptides and Proteins
  • MAGP2 protein, human
  • Protein Precursors
  • Proteins
  • microfibrillar protein
  • proenkephalin
  • Interleukin-10