{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,2]],"date-time":"2026-09-02T11:30:35Z","timestamp":1788348635664,"version":"build-2803163510"},"reference-count":46,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2022,11,17]],"date-time":"2022-11-17T00:00:00Z","timestamp":1668643200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Dutch Technology Foundation TTW","award":["NWO-VIDI 12950"],"award-info":[{"award-number":["NWO-VIDI 12950"]}]},{"name":"Dutch Technology Foundation TTW","award":["2099-196678"],"award-info":[{"award-number":["2099-196678"]}]},{"name":"Ministry of Economic Affairs","award":["NWO-VIDI 12950"],"award-info":[{"award-number":["NWO-VIDI 12950"]}]},{"name":"Ministry of Economic Affairs","award":["2099-196678"],"award-info":[{"award-number":["2099-196678"]}]},{"name":"de Cock-Hadders Stichting","award":["NWO-VIDI 12950"],"award-info":[{"award-number":["NWO-VIDI 12950"]}]},{"name":"de Cock-Hadders Stichting","award":["2099-196678"],"award-info":[{"award-number":["2099-196678"]}]},{"name":"Erasmus Mundus India\u2013EU mobility consortium","award":["NWO-VIDI 12950"],"award-info":[{"award-number":["NWO-VIDI 12950"]}]},{"name":"Erasmus Mundus India\u2013EU mobility consortium","award":["2099-196678"],"award-info":[{"award-number":["2099-196678"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Pharmaceutics"],"abstract":"<jats:p>Cell-derived extracellular vesicles (EVs) are effectors of cell-to-cell communication that are in the spotlight as promising candidates for in vivo drug delivery because of their ability to enter cells and deliver cargo. For example, proteins of interest can be loaded into EVs to mediate protein transfer into target cells. To determine causality between EV content and function, which is also important to assess the clinical safety of EVs, it is crucial to comprehensively characterize their complete molecular composition. Here, we investigated EVs loaded with the chaperone protein DNAJB6. Chaperone proteins assist in protein folding and have been suggested to alleviate protein aggregation diseases, such as Alzheimer\u2019s disease and Huntington\u2019s disease. We analyzed and compared the proteome of EVs isolated from wildtype HEK293T cells with that of EVs from HEK 293T cells overexpressing DNAJB6-WT or loss-of-function mutant DNAJB6-M3. Comprehensive analysis of proteomics data showed enhanced levels of DNAJB6 as well as protein-folding-related proteins in EVs derived from DNAJB6-overexpression cells. Interestingly, upregulation of a chaperone and its protein-folding-related proteins resulted in downregulation of another chaperone plus its related proteins, and vice versa. This implies the presence of compensatory mechanisms in the cellular expression of chaperones. Collectively, we provide the proteomic EV signatures underlying EV mediated DNAJB6 transmission by HEK293T cells, with the aim of establishing a causal relationship between EV protein content and EV function.<\/jats:p>","DOI":"10.3390\/pharmaceutics14112485","type":"journal-article","created":{"date-parts":[[2022,11,17]],"date-time":"2022-11-17T03:26:04Z","timestamp":1668655564000},"page":"2485","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["DNAJB6-Containing Extracellular Vesicles as Chaperone Delivery Systems: A Proteomic Analysis"],"prefix":"10.3390","volume":"14","author":[{"given":"Bhagyashree S.","family":"Joshi","sequence":"first","affiliation":[{"name":"Department of Biomedical Engineering, University Medical Center Groningen, University of Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Hector","family":"Garcia Romeu","sequence":"additional","affiliation":[{"name":"Groningen Research Institute of Pharmacy, University Medical Center Groningen, University of Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Aldy","family":"Aliyandi","sequence":"additional","affiliation":[{"name":"Groningen Research Institute of Pharmacy, University Medical Center Groningen, University of Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marcel P.","family":"de Vries","sequence":"additional","affiliation":[{"name":"Interfaculty Mass Spectrometry Center, University Medical Center Groningen, University of Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7695-915X","authenticated-orcid":false,"given":"Inge S.","family":"Zuhorn","sequence":"additional","affiliation":[{"name":"Department of Biomedical Engineering, University Medical Center Groningen, University of Groningen, Antonius Deusinglaan 1, 9713AV Groningen, The Netherlands"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2022,11,17]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"1226","DOI":"10.1016\/j.cell.2016.01.043","article-title":"Communication by Extracellular Vesicles: Where We Are and Where We Need to Go","volume":"164","author":"Tkach","year":"2016","journal-title":"Cell"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"27066","DOI":"10.3402\/jev.v4.27066","article-title":"Biological properties of extracellular vesicles and their physiological functions","volume":"4","author":"Siljander","year":"2015","journal-title":"J. Extracell. Vesicles"},{"key":"ref_3","first-page":"203","article-title":"Emerging roles of exosomes in normal and pathological conditions: New insights for diagnosis and therapeutic applications","volume":"6","author":"Herschlik","year":"2015","journal-title":"Front. Immunol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"6478","DOI":"10.1126\/science.aau6977","article-title":"The biology, function, and biomedical applications of exosomes","volume":"367","author":"Kalluri","year":"2020","journal-title":"Science"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"329","DOI":"10.1038\/nature15756","article-title":"Tumour exosome integrins determine organotropic metastasis","volume":"527","author":"Hoshino","year":"2015","journal-title":"Nature"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1305","DOI":"10.1038\/s41467-018-03733-8","article-title":"Designer exosomes produced by implanted cells intracerebrally deliver therapeutic cargo for Parkinson\u2019s disease treatment","volume":"9","author":"Kojima","year":"2018","journal-title":"Nat. Commun."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1269","DOI":"10.1016\/j.ymthe.2017.03.030","article-title":"Engineered Exosomes as Vehicles for Biologically Active Proteins","volume":"25","author":"Sterzenbach","year":"2017","journal-title":"Mol. Ther."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"1133","DOI":"10.1158\/1535-7163.MCT-17-0827","article-title":"Anti-HER2 scFv-Directed Extracellular Vesicle-Mediated mRNA-Based Gene Delivery Inhibits Growth of HER2-Positive Human Breast Tumor Xenografts by Prodrug Activation","volume":"17","author":"Wang","year":"2018","journal-title":"Mol. Cancer Ther."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"18","DOI":"10.1016\/j.jconrel.2015.03.033","article-title":"Exosomes as drug delivery vehicles for Parkinson\u2019s disease therapy","volume":"207","author":"Haney","year":"2015","journal-title":"J. Control. Release"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"e1904040","DOI":"10.1002\/adma.201904040","article-title":"Exosomes and Nanoengineering: A Match Made for Precision Therapeutics","volume":"32","author":"Tran","year":"2020","journal-title":"Adv. Mater."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"16086","DOI":"10.1073\/pnas.1902513116","article-title":"Exosomes regulate neurogenesis and circuit assembly","volume":"116","author":"Sharma","year":"2019","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"72","DOI":"10.1016\/j.jconrel.2014.07.049","article-title":"Extracellular vesicles as drug delivery systems: Lessons from the liposome field","volume":"195","author":"Fens","year":"2014","journal-title":"J. Control. Release"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"1761","DOI":"10.1021\/acs.accounts.9b00109","article-title":"Drug Delivery with Extracellular Vesicles: From Imagination to Innovation","volume":"52","author":"Kooijmans","year":"2019","journal-title":"Acc. Chem. Res."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"E2497","DOI":"10.1073\/pnas.1412651112","article-title":"Intercellular chaperone transmission via exosomes contributes to maintenance of protein homeostasis at the organismal level","volume":"112","author":"Takeuchi","year":"2015","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"108","DOI":"10.1016\/j.nbd.2018.10.017","article-title":"Astrocytic expression of the chaperone DNAJB6 results in non-cell autonomous protection in Huntington\u2019s disease","volume":"124","author":"Bason","year":"2019","journal-title":"Neurobiol. Dis."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"1332","DOI":"10.1038\/s41593-018-0235-9","article-title":"Protein misfolding, aggregation, and conformational strains in neurodegenerative diseases","volume":"21","author":"Soto","year":"2018","journal-title":"Nat. Neurosci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1002\/mas.21420","article-title":"Proteomics of extracellular vesicles: Exosomes and ectosomes","volume":"34","author":"Choi","year":"2015","journal-title":"Mass Spectrom Rev."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"32570","DOI":"10.3402\/jev.v5.32570","article-title":"High-resolution proteomic and lipidomic analysis of exosomes and microvesicles from different cell sources","volume":"5","author":"Haraszti","year":"2016","journal-title":"J. Extracell. Vesicles"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"10813","DOI":"10.1038\/s41598-018-28485-9","article-title":"Heterogeneity and interplay of the extracellular vesicle small RNA transcriptome and proteome","volume":"8","author":"Sork","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_20","doi-asserted-by":"crossref","unstructured":"Joshi, B.S., and Zuhorn, I.S. (2022). Preparation of chaperone protein-loaded neural stem cell-derived extracellular vesicles to reduce protein aggregation in cellular models of Huntington\u2019s disease. STAR Protoc.","DOI":"10.1016\/j.xpro.2023.102134"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"1367","DOI":"10.1038\/nbt.1511","article-title":"MaxQuant enables high peptide identification rates, individualized p.p.b.-range mass accuracies and proteome-wide protein quantification","volume":"26","author":"Cox","year":"2008","journal-title":"Nat. Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1093\/nar\/gkn923","article-title":"Bioinformatics enrichment tools: Paths toward the comprehensive functional analysis of large gene lists","volume":"37","author":"Sherman","year":"2009","journal-title":"Nucleic Acids Res."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"44","DOI":"10.1038\/nprot.2008.211","article-title":"Systematic and integrative analysis of large gene lists using DAVID bioinformatics resources","volume":"4","author":"Sherman","year":"2009","journal-title":"Nat. Protoc."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"420","DOI":"10.1038\/s41587-019-0036-z","article-title":"SignalP 5.0 improves signal peptide predictions using deep neural networks","volume":"37","author":"Tsirigos","year":"2019","journal-title":"Nat. Biotechnol."},{"key":"ref_25","unstructured":"Oliveros, J.C. (2022, October 01). VENNY. An Interactive Tool for Comparing Lists with Venn Diagrams. Available online: http:\/\/bioinfogp.cnb.csic.es\/tools\/venny\/index.html."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"272","DOI":"10.1016\/j.molcel.2016.03.017","article-title":"The S\/T-Rich Motif in the DNAJB6 Chaperone Delays Polyglutamine Aggregation and the Onset of Disease in a Mouse Model","volume":"62","author":"Kakkar","year":"2016","journal-title":"Mol. Cell"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1016\/j.molcel.2010.01.001","article-title":"A DNAJB chaperone subfamily with HDAC-dependent activities suppresses toxic protein aggregation","volume":"37","author":"Hageman","year":"2010","journal-title":"Mol. Cell"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"R22","DOI":"10.1186\/bcr1874","article-title":"Large isoform of MRJ (DNAJB6) reduces malignant activity of breast cancer","volume":"10","author":"Mitra","year":"2008","journal-title":"Breast Cancer Res."},{"key":"ref_29","doi-asserted-by":"crossref","unstructured":"Pei, Y., Fu, W., Yang, E., Shen, A., Chen, Y.C., Gong, H., Chen, J., Huang, J., Xiao, G., and Liu, F. (2012). A Hsp40 chaperone protein interacts with and modulates the cellular distribution of the primase protein of human cytomegalovirus. PLoS Pathog., 8.","DOI":"10.1371\/journal.ppat.1002968"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"149","DOI":"10.1023\/A:1024916223616","article-title":"Characterization of two isoforms of a human DnaJ homologue, HSJ2","volume":"30","author":"Hanai","year":"2003","journal-title":"Mol. Biol. Rep."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"9039","DOI":"10.1038\/s41598-017-08324-z","article-title":"The molecular chaperones DNAJB6 and Hsp70 cooperate to suppress alpha-synuclein aggregation","volume":"7","author":"Aprile","year":"2017","journal-title":"Sci. Rep."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"18374","DOI":"10.3402\/jev.v1i0.18374","article-title":"ExoCarta as a resource for exosomal research","volume":"1","author":"Simpson","year":"2012","journal-title":"J. Extracell. Vesicles"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"869","DOI":"10.1096\/fj.01-0995fje","article-title":"Preferential transformation of human neuronal cells by human adenoviruses and the origin of HEK 293 cells","volume":"16","author":"Shaw","year":"2002","journal-title":"FASEB J."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"4767","DOI":"10.1038\/ncomms5767","article-title":"Genome dynamics of the human embryonic kidney 293 lineage in response to cell biology manipulations","volume":"5","author":"Lin","year":"2014","journal-title":"Nat. Commun."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"2840","DOI":"10.1093\/bioinformatics\/btu383","article-title":"Literome: PubMed-scale genomic knowledge base in the cloud","volume":"30","author":"Poon","year":"2014","journal-title":"Bioinformatics"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"jcs227033","DOI":"10.1242\/jcs.227033","article-title":"DnaJB6 is a RanGTP-regulated protein required for microtubule organization during mitosis","volume":"132","author":"Scrofani","year":"2019","journal-title":"J. Cell Sci."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"675","DOI":"10.1007\/s10495-016-1244-3","article-title":"Clinical and pathological significance of N-Myc downstream-regulated gene 2 (NDRG2) in diverse human cancers","volume":"21","author":"Hu","year":"2016","journal-title":"Apoptosis"},{"key":"ref_38","first-page":"D433","article-title":"STRING: Known and predicted protein-protein associations, integrated and transferred across organisms","volume":"33","author":"Jensen","year":"2005","journal-title":"Nucleic Acids Res."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"569","DOI":"10.1038\/nri855","article-title":"Exosomes: Composition, biogenesis and function","volume":"2","author":"Thery","year":"2002","journal-title":"Nat. Rev. Immunol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"940","DOI":"10.1016\/j.bbagen.2012.03.017","article-title":"Exosomes: Current knowledge of their composition, biological functions, and diagnostic and therapeutic potentials","volume":"1820","author":"Vlassov","year":"2012","journal-title":"Biochim. Biophys. Acta"},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"133","DOI":"10.1016\/j.molcel.2010.06.010","article-title":"Secreted monocytic miR-150 enhances targeted endothelial cell migration","volume":"39","author":"Zhang","year":"2010","journal-title":"Mol. Cell"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"883","DOI":"10.1038\/nm.2753","article-title":"Melanoma exosomes educate bone marrow progenitor cells toward a pro-metastatic phenotype through MET","volume":"18","author":"Peinado","year":"2012","journal-title":"Nat. Med."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"816","DOI":"10.1038\/ncb3169","article-title":"Pancreatic cancer exosomes initiate pre-metastatic niche formation in the liver","volume":"17","author":"Aiello","year":"2015","journal-title":"Nat. Cell Biol."},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"310","DOI":"10.1016\/j.addr.2009.02.003","article-title":"Induction of heat shock proteins for protection against oxidative stress","volume":"61","author":"Kalmar","year":"2009","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"317","DOI":"10.5114\/ceji.2016.63133","article-title":"Heat shock proteins (HSPs) in the homeostasis of regulatory T cells (Tregs)","volume":"41","author":"Kolinski","year":"2016","journal-title":"Cent. Eur. J. Immunol."},{"key":"ref_46","doi-asserted-by":"crossref","unstructured":"Lee, S., Mankhong, S., and Kang, J.H. (2019). Extracellular Vesicle as a Source of Alzheimer\u2019s Biomarkers: Opportunities and Challenges. Int. J. Mol. Sci., 20.","DOI":"10.3390\/ijms20071728"}],"container-title":["Pharmaceutics"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1999-4923\/14\/11\/2485\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T01:19:52Z","timestamp":1760145592000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1999-4923\/14\/11\/2485"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2022,11,17]]},"references-count":46,"journal-issue":{"issue":"11","published-online":{"date-parts":[[2022,11]]}},"alternative-id":["pharmaceutics14112485"],"URL":"https:\/\/doi.org\/10.3390\/pharmaceutics14112485","relation":{},"ISSN":["1999-4923"],"issn-type":[{"value":"1999-4923","type":"electronic"}],"subject":[],"published":{"date-parts":[[2022,11,17]]}}}