title: Tracing Cellular Origin of Human Exosomes Using Multiplex Proximity Extension Assays.
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.
authors.affiliation: From the ‡Department of Medicine, Unit for Immunology and Allergy, Karolinska Institutet and Karolinska University Hospital, SE-171 76 Stockholm, Sweden.
authors.forename: Pia
authors.initials: P
authors.lastname: Larssen
authors.affiliation: §Department of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, SE-751 08 Uppsala, Sweden.
authors.forename: Lotta
authors.initials: L
authors.lastname: Wik
authors.affiliation: ¶Department of Medicine, Unit for Immunology and Allergy, Science for Life Laboratory, Karolinska Institutet and Karolinska University Hospital, SE-171 76 Stockholm, Sweden.
authors.forename: Paulo
authors.initials: P
authors.lastname: Czarnewski
authors.affiliation: From the ‡Department of Medicine, Unit for Immunology and Allergy, Karolinska Institutet and Karolinska University Hospital, SE-171 76 Stockholm, Sweden.
authors.forename: Maria
authors.initials: M
authors.lastname: Eldh
authors.affiliation: §Department of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, SE-751 08 Uppsala, Sweden.
authors.forename: Liza
authors.initials: L
authors.lastname: Löf
authors.affiliation: ‖Department of Medical Sciences, Clinical Chemistry, Uppsala University, SE-751 85 Uppsala, Sweden.
authors.forename: K Göran
authors.initials: KG
authors.lastname: Ronquist
authors.affiliation: ‖Department of Medical Sciences, Clinical Chemistry, Uppsala University, SE-751 85 Uppsala, Sweden.
authors.forename: Louise
authors.initials: L
authors.lastname: Dubois
authors.affiliation: **Department of Medical Sciences, Cancer Pharmacology and Computational Medicine, National Bioinformatics Infrastructure Sweden, Science for Life Laboratory, Uppsala University, SE-751 85 Uppsala, Sweden.
authors.forename: Eva
authors.initials: E
authors.lastname: Freyhult
authors.affiliation: §Department of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, SE-751 08 Uppsala, Sweden.
authors.forename: Caroline J
authors.initials: CJ
authors.lastname: Gallant
authors.affiliation: §Department of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, SE-751 08 Uppsala, Sweden.
authors.forename: Johan
authors.initials: J
authors.lastname: Oelrich
authors.affiliation: ‖Department of Medical Sciences, Clinical Chemistry, Uppsala University, SE-751 85 Uppsala, Sweden.
authors.forename: Anders
authors.initials: A
authors.lastname: Larsson
authors.affiliation: ‖Department of Medical Sciences, Clinical Chemistry, Uppsala University, SE-751 85 Uppsala, Sweden.
authors.forename: Gunnar
authors.initials: G
authors.lastname: Ronquist
authors.affiliation: ¶Department of Medicine, Unit for Immunology and Allergy, Science for Life Laboratory, Karolinska Institutet and Karolinska University Hospital, SE-171 76 Stockholm, Sweden.
authors.forename: Eduardo J
authors.initials: EJ
authors.lastname: Villablanca
authors.affiliation: §Department of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, SE-751 08 Uppsala, Sweden.
authors.forename: Ulf
authors.initials: U
authors.lastname: Landegren
authors.affiliation: From the ‡Department of Medicine, Unit for Immunology and Allergy, Karolinska Institutet and Karolinska University Hospital, SE-171 76 Stockholm, Sweden.
authors.forename: Susanne
authors.initials: S
authors.lastname: Gabrielsson
authors.affiliation: §Department of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, SE-751 08 Uppsala, Sweden; masood.kamali@igp.uu.se.
authors.forename: Masood
authors.initials: M
authors.lastname: Kamali-Moghaddam
mesh_terms: D015415:Biomarkers / Q000378:metabolism*; D001826:Body Fluids / Q000378:metabolism*; D002460:Cell Line; D055252:Cell-Derived Microparticles / Q000378:metabolism*; D055354:Exosomes / Q000378:metabolism*; D005260:Female; D057166:High-Throughput Screening Assays / Q000379:methods*; D006801:Humans; D020014:K562 Cells; D061986:MCF-7 Cells; D008297:Male; D008895:Milk, Human / Q000378:metabolism; D009928:Organ Specificity; D025341:Principal Component Analysis; D011467:Prostate / Q000378:metabolism
chemical_list: D015415:Biomarkers