Larssen Pia, Wik Lotta, Czarnewski Paulo, Eldh Maria, Löf Liza, Ronquist K Göran, Dubois Louise, Freyhult Eva, Gallant Caroline J, Oelrich Johan, Larsson Anders, Ronquist Gunnar, Villablanca Eduardo J, Landegren Ulf, Gabrielsson Susanne, Kamali-Moghaddam Masood
From the ‡Department of Medicine, Unit for Immunology and Allergy, Karolinska Institutet and Karolinska University Hospital, SE-171 76 Stockholm, Sweden.
§Department of Immunology, Genetics & Pathology, Science for Life Laboratory, Uppsala University, SE-751 08 Uppsala, Sweden.
Mol Cell Proteomics. 2017 Mar;16(3):502-511. doi: 10.1074/mcp.M116.064725. Epub 2017 Jan 22.
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.
细胞外囊泡（EVs）是由多种细胞类型释放的膜包被结构，如外泌体和微囊泡。它们在体液中的存在以及可变的表面组成和内容物使其成为有吸引力的潜在生物标志物。确定其细胞来源的能力可以极大地推动该领域的发展。我们使用多重邻近延伸分析（PEA）以高特异性和敏感性鉴定不同来源外泌体的蛋白质谱，包括七种细胞系和两种不同的体液。通过比较细胞和外泌体，我们成功鉴定了产生外泌体的细胞。此外，通过对蛋白质模式的主成分分析，人乳EVs和前列腺腺泡细胞释放的前列腺小体分别与来自乳腺和前列腺组织的细胞系聚类。乳外泌体独特地表达CXCL5、MIA和KLK6，而前列腺小体携带NKX31、GSTP1和SRC，这突出表明来自不同来源的EVs表达不同的蛋白质。总之，PEA为外泌体研究提供了一种强大的蛋白质筛选工具，用于鉴定外泌体的细胞来源或癌症和炎症等疾病中的新生物标志物。
