title: Comprehensive Proteomic Analysis of Mesenchymal Stem Cell Exosomes Reveals Modulation of Angiogenesis via Nuclear Factor-KappaB Signaling.
abstract: Mesenchymal stem cells (MSC) are known to facilitate healing of ischemic tissue related diseases through proangiogenic secretory proteins. Recent studies further show that MSC derived exosomes function as paracrine effectors of angiogenesis, however, the identity of which components of the exosome proteome responsible for this effect remains elusive. To address this we used high-resolution isoelectric focusing coupled liquid chromatography tandem mass spectrometry, an unbiased high throughput proteomics approach to comprehensively characterize the proteinaceous contents of MSCs and MSC derived exosomes. We probed the proteome of MSCs and MSC derived exosomes from cells cultured under expansion conditions and under ischemic tissue simulated conditions to elucidate key angiogenic paracrine effectors present and potentially differentially expressed in these conditions. In total, 6,342 proteins were identified in MSCs and 1,927 proteins in MSC derived exosomes, representing to our knowledge the first time these proteomes have been probed comprehensively. Multilayered analyses identified several putative paracrine effectors of angiogenesis present in MSC exosomes and increased in expression in MSCs exposed to ischemic tissue-simulated conditions; these include platelet derived growth factor, epidermal growth factor, fibroblast growth factor, and most notably nuclear factor-kappaB (NFkB) signaling pathway proteins. NFkB signaling was identified as a key mediator of MSC exosome induced angiogenesis in endothelial cells by functional in vitro validation using a specific inhibitor. Collectively, the results of our proteomic analysis show that MSC derived exosomes contain a robust profile of angiogenic paracrine effectors, which have potential for the treatment of ischemic tissue-related diseases.
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Johnathon D
authors.initials: JD
authors.lastname: Anderson
authors.affiliation: Cancer Proteomics, Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Henrik J
authors.initials: HJ
authors.lastname: Johansson
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Calvin S
authors.initials: CS
authors.lastname: Graham
authors.affiliation: Cancer Proteomics, Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Mattias
authors.initials: M
authors.lastname: Vesterlund
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Missy T
authors.initials: MT
authors.lastname: Pham
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Charles S
authors.initials: CS
authors.lastname: Bramlett
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Elizabeth N
authors.initials: EN
authors.lastname: Montgomery
authors.affiliation: Surgical and Radiological Sciences, Department of Veterinary Medicine, University of California Davis, Davis, California, USA.
authors.forename: Matt S
authors.initials: MS
authors.lastname: Mellema
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Renee L
authors.initials: RL
authors.lastname: Bardini
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Zelenia
authors.initials: Z
authors.lastname: Contreras
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Madeline
authors.initials: M
authors.lastname: Hoon
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Gerhard
authors.initials: G
authors.lastname: Bauer
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Kyle D
authors.initials: KD
authors.lastname: Fink
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Brian
authors.initials: B
authors.lastname: Fury
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Kyle J
authors.initials: KJ
authors.lastname: Hendrix
authors.affiliation: Department of Molecular and Cellular Biology, University of California Davis, Davis, California, USA.
authors.forename: Frederic
authors.initials: F
authors.lastname: Chedin
authors.affiliation: Department of Laboratory Medicine, Karolinska Institutet, Stockholm, Sweden.|Department of Physiology, Anatomy and Genetics, University of Oxford, Oxford, United Kingdom.
authors.forename: Samir
authors.initials: S
authors.lastname: El-Andaloussi
authors.affiliation: Department of Surgery, University of Washington, Seattle, Washington, USA.
authors.forename: Billie
authors.initials: B
authors.lastname: Hwang
authors.affiliation: Department of Surgery, University of Washington, Seattle, Washington, USA.
authors.forename: Michael S
authors.initials: MS
authors.lastname: Mulligan
authors.affiliation: Cancer Proteomics, Department of Oncology-Pathology, Karolinska Institutet, Stockholm, Sweden.
authors.forename: Janne
authors.initials: J
authors.lastname: Lehtiö
authors.affiliation: Stem Cell Program, Department of Internal Medicine, University of California Davis, Davis, California, USA.
authors.forename: Jan A
authors.initials: JA
authors.lastname: Nolta
mesh_terms: D001854:Bone Marrow Cells / Q000378:metabolism; D055354:Exosomes / Q000235:genetics*; D018507:Gene Expression Regulation, Developmental; D006801:Humans; D008297:Male; D059630:Mesenchymal Stem Cells / Q000378:metabolism*; D016328:NF-kappa B / Q000235:genetics*; D018919:Neovascularization, Physiologic / Q000235:genetics*; D019899:Paracrine Communication / Q000235:genetics; D020543:Proteome / Q000235:genetics; D015398:Signal Transduction; D055815:Young Adult
chemical_list: D016328:NF-kappa B; D020543:Proteome
keywords: Exosomes; High-resolution isoelectric focusing; Liquid chromatography tandem mass spectrometry; Mesenchymal stem cells; Nuclear factor kappaB; Peripheral arterial disease; Proteomics