Enriched protein screening of human bone marrow mesenchymal stromal cell secretions reveals MFAP5 and PENK as novel IL-10 modulators.
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.
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.
Jack M
JM
Milwid
Department of Surgery, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Jessica S
JS
Elman
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.
Matthew
M
Li
Department of Surgery, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Keyue
K
Shen
Harvard-MIT Division of Health Sciences and Technology, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Arjun
A
Manrai
Computational Biology Center, Memorial Sloan-Kettering Cancer Center, New York, New York, USA.
Aaron
A
Gabow
Department of Biomedical Engineering, Rutgers University, Piscataway, New Jersey, USA.
Joshua
J
Yarmush
Department of Surgery, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Yunxin
Y
Jiao
Dana-Farber Cancer Institute, Harvard Medical School, Boston, Massachusetts, USA.
Anne
A
Fletcher
Department of Surgery, Center for Engineering in Medicine and Surgical Services, Massachusetts General Hospital, Harvard Medical School, Boston, Massachusetts, USA.
Jungwoo
J
Lee
David H. Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA.
Michael J
MJ
Cima
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.
Martin L
ML
Yarmush
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.
Biju
B
Parekkadan
D000818:Animals; D001854:Bone Marrow Cells / Q000276:immunology / Q000378:metabolism; D003285:Contractile Proteins / Q000235:genetics* / Q000378:metabolism*; D004745:Enkephalins / Q000235:genetics* / Q000378:metabolism; D016326:Extracellular Matrix Proteins / Q000235:genetics / Q000378:metabolism*; D020869:Gene Expression Profiling; D006023:Glycoproteins / Q000235:genetics*; D006801:Humans; D036341:Intercellular Signaling Peptides and Proteins; D016753:Interleukin-10 / Q000235:genetics / Q000378:metabolism*; D059630:Mesenchymal Stem Cells / Q000276:immunology / Q000378:metabolism; D051379:Mice; D014176:Protein Biosynthesis / Q000235:genetics; D011498:Protein Precursors / Q000235:genetics* / Q000378:metabolism; D011506:Proteins / Q000235:genetics / Q000378:metabolism*; D040901:Proteomics; D000072260:RNA Splicing Factors; D012772:Shock, Septic / Q000235:genetics / Q000628:therapy*
D003285:Contractile Proteins; D004745:Enkephalins; D016326:Extracellular Matrix Proteins; D006023:Glycoproteins; C508609:IL10 protein, human; D036341:Intercellular Signaling Peptides and Proteins; C115627:MFAP5 protein, human; D011498:Protein Precursors; D011506:Proteins; D000072260:RNA Splicing Factors; C016598:microfibrillar protein; C029992:proenkephalin; D016753:Interleukin-10