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

Jack M Milwid; Jessica S Elman; Matthew Li; Keyue Shen; Arjun Manrai; Aaron Gabow; Joshua Yarmush; Yunxin Jiao; Anne Fletcher; Jungwoo Lee; Michael J Cima; Martin L Yarmush; Biju Parekkadan

doi:10.1038/mt.2014.17

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.

Authors

Jack M Milwid¹, Jessica S Elman², Matthew Li³, Keyue Shen², Arjun Manrai⁴, Aaron Gabow⁵, Joshua Yarmush⁶, Yunxin Jiao², Anne Fletcher⁷, Jungwoo Lee², Michael J Cima⁸, Martin L Yarmush⁹, Biju Parekkadan¹⁰

Affiliations

¹ 1] Department of Surgery, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA [2] Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
² Department of Surgery, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
³ 1] Department of Surgery, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA [2] Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA [3] David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
⁴ Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
⁵ Computational Biology Center, Memorial Sloan-Kettering Cancer Center, New York, New York, USA.
⁶ Department of Biomedical Engineering, Rutgers University, Piscataway, New Jersey, USA.
⁷ Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, USA.
⁸ David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
⁹ 1] Department of Surgery, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA [2] Department of Biomedical Engineering, Rutgers University, Piscataway, New Jersey, USA.
¹⁰ 1] Department of Surgery, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA [2] Harvard Stem Cell Institute, Cambridge, Massachusetts, USA.

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
RNA Splicing Factors
Shock, Septic / genetics
Shock, Septic / therapy*

Substances

Contractile Proteins
Enkephalins
Extracellular Matrix Proteins
Glycoproteins
IL10 protein, human
Intercellular Signaling Peptides and Proteins
MFAP5 protein, human
Protein Precursors
Proteins
RNA Splicing Factors
microfibrillar protein
proenkephalin
Interleukin-10

Abstract

Publication types

MeSH terms

Substances

Grants and funding