A. Agazzi

A. Buness

F. Shakeri

G. Savoini

H. Sauerwein

M. Sylvester

R. Furioso Ferreira

T. Blees

V. Mrljak

English

AIR Universita degli studi di Milano

DOI: !".!""#/jev#.!#"$%ISEV!"!# Abstract BookAbout ISEVThe International Society for Extracellular Vesicles is the leading professional society for researchers and scientists involved inthe study of microvesicles and exosomes. With nearly !,"""members, ISEV continues to be the leader in advancing the study ofextracellular vesicles. Founded in #"!# in Sweden, ISEV has since moved its Headquarters to the United States. Through its pro-grams and services, ISEV provides essential training and research opportunities for those involved in exosome and microvesicleresearch.Mission StatementAdvancing extracellular vesicle research globally.VisionOur vision is to be the leading advocate and guide of extracellular vesicle research and to advance the understanding of extracel-lular vesicle biology.ISEV!"!# Annual MeetingThe International Society for Extracellular Vesicles is the is the premier international conference of extracellular vesicle research,covering the latest in exosomes, microvesicles andmore.With an anticipated !,""" attendees, ISEV#"#!will feature presentationsfrom the top researchers in the &eld, as well as providing opportunities for talks from students and early career researchers.ISEV!"!# International Organizing CommitteeIOC Chairs: Lorraine O’Driscoll (Ireland), Sophie Rome (France)IOCMembers: Antonella Bongiovanni (Italy), DaveCarter (UnitedKingdom), VincentHyenne (France), Soazig Le Lay (France),Andreas Möller (New Zealand), Eva Rohde (Austria), Tang-Long Shen (Taiwan), Carolina Soekmadji (Australia), and KenWitwer (USA)Journal of Extracellular Vesicles: Editors in ChiefJan Lotvall (Sweden)This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the originalwork is properly cited.© #"#! The Authors. Journal of Extracellular Vesicles published by Wiley Periodicals, LLC on behalf of the International Society for Extracellular VesiclesISEV!"!# A$%&'()& B**+ !"# of !$%PS&$.&% Tumor-derived microRNA-'#(a-'p-containing extracellular vesicles induceosteoclastogenesis by activating Dyrk&a/Nfatc&/Angptl! axis to promote the metastasis of prostatecancerJialing Wang, State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren JiHospital, School of Medicine, Shanghai Jiao Tong UniversityYanqing Wang, Department of Urology, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityBaijun Dong, Department of Urology, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityWei Xue, Department of Urology, Renji Hospital, School of Medicine, Shanghai Jiao Tong UniversityWei-Qiang Gao, State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, RenJi Hospital, School of Medicine, Shanghai Jiao Tong UniversityYu-Xiang Fang, State Key Laboratory of Oncogenes and Related Genes, Renji-Med X Clinical Stem Cell Research Center, Ren JiHospital, School of Medicine, Shanghai Jiao Tong UniversityIntroduction: Themajority of the deaths of prostate cancer (PCa) are caused by progression to bonemetastatic PCa. The impor-tance of extracellular vesicles (EVs) in the formation of the pre-metastatic niche has been demonstrated in recent years. However,whether and how tumor-derived EVs interact with pre-osteoclasts to release EV-delivered microRNAs to activate pre-metastaticniche formation and prostate cancer bone metastasis remain unclear.Methods: Bioinformatics and qRT-PCR analyses were used to screen and identify the expression of miR-,-.a-,p in both serumand tissue derived EVs fromprimary ormetastatic PCa patients. Biological function assay studies in vitro and in vivo were imple-mented to identify the functions of miR-,-.a-,p during PCa progression. Dual-luciferase reporter assay, co-IP assay, westernblot assay, IF staining, RIP and ChIP assays were conducted to investigate the underlying mechanism.Results:We found that EV-mediated release of miR-,-.a-,p from tumor cells is upregulated in bone-metastatic PCa for mainte-nance of a low intercellular concentration of miR-,-.a-,p to promote proliferation and anti-apoptosis of PCa cells and aMAOA-mediated epithelial-to-mesenchymal transition (EMT) formigration as well. Furthermore, we demonstrated that the enrichmentofmiR-,-.a-,p in tumor delivered EVswas induced by overexpression of hnRNPA!B#, a RNAbinding protein, as a transfer chap-erone. After miR-,-.a-,p enriched EVs was absorbed by pre-osteoclasts, elevated intercellular concentration of miR-,-.a-,p inpre-osteoclasts promotes osteoclastogenesis by targeting Dyrk#a/Nfatc# signaling pathway. Moreover, inhibition of Dyrk#a bymiR-,-.a-,p attenuated its sequestration to Nfatc# so that the nuclear translocation of Nfatc# was increased to promote expres-sion of the downstream target gene Angptl!. As a feedback, increased secret of Angptl! into the environment was found to becapable to improve PCa progression.Summary/Conclusion: Our /ndings indicate that tumor-derived miR-,-.a-,p-containing EVs plays a signi/cant role in pro-moting prostate cancer bonemetastasis by activating Dyrk#a/Nfatc#/Angptl! axis in pre-osteoclasts to induce osteoclastogenesis,which implicates that miR-,-.a-,p may be a potential predictor of metastatic PCa. Moreover, reducing the release of miR-,-.a-,p-containing EVs or blocking the function of miR-,-.a-,p in pre-osteoclasts might be a potential therapeutic strategy for PCametastasis.PS!) EVs and Our Non-Human FriendsChair: Kwang Pyo Kim, Department of Applied Chemistry, Institute of Natural Science, Global Center for Pharmaceutical Ingre-dient Materials, Kyung Hee University, Yongin, South Korea; Department of Biomedical Science and Technology, Kyung HeeMedical Science Research Institute, Kyung Hee University, Seoul, South KoreaChair: Zheng Lei, Department of Laboratory Medicine, Nanfang Hospital, SouthernMedical University, China (People’s Repub-lic)PS!).)& Comparative proteome pro*ling in exosomes derived from porcine colostrum versusmature milkRafaela Furioso Ferreira, Institute of Animal Science, Physiology Unit, University of BonnThomas Blees, Institute of Animal Science & Physiology Unit, University of BonnFarhad Shakeri, Institute for Medical Biometry, Informatics and Epidemiology, Medical Faculty, University of BonnAndreas Buness, Institute for Medical Biometry, Informatics and Epidemiology, Medical Faculty, University of BonnMarc Sylvester, Institute of Biochemistry and Molecular Biology, Core Facility Mass Spectrometry, Medical Faculty, Universityof BonnAlessandro Agazzi, Department of Health, Animal Science and Food Safety ‘Carlo Cantoni’ (VESPA), University of Milan!"# of !$% ISEV!"!# A$%&'()& B**+Giovanni Savoini, Department of Health, Animal Science and Food Safety ‘Carlo Cantoni’ (VESPA), University of MilanVladimir Mrljak, Faculty of Veterinary Medicine, University of ZagrebHelga Sauerwein, Institute of Animal Science & Physiology Unit, University of BonnIntroduction: Colostrum and milk have high nutritional value and provide a complete diet for neonates, along with bioac-tive substances which modulate various functions such as immune defense. The mechanisms by which milk components canprime the infant’s active immunity are not entirely clear, and EVs are suggested to be essential for the infant’s physiologicaldevelopment.Methods: We assessed the exosomal proteome pro,le from milk samples obtained from #" healthy sows, at day " (colostrum),day -, and #. post partum. Exosomes were isolated by ultracentrifugation coupled with size exclusion chromatography, and werecharacterized by nanoparticle tracking analysis, transmission electron microscopy and Western blotting for exosome markers.Isolated exosomeswere in-gel digested and after TMT-labelling of the peptides, theywere subjected to LC-MS/MS. The statisticalanalyses were performed in R using an in-house developed work/ow. Non-unique peptides, single-hit proteins, and fractionswith low intensity per protein were ,ltered out. The data were transformed and normalized using the VSN package and thenaggregated to protein-level by Tukey’s median polish procedure. The P-values were adjusted for multiple testing by Benjamini-Hochberg method.Results:After exclusion criteria were applied, a total of 0#1 proteins in each timepoint were statistically analyzed. Exosomes fromcolostrum presented #2! di3erentially abundant proteins (DAP) (4! increased and 4" decreased) as compared to exosomes frommilk at day -, and #-"DAP (4# increased and 41 decreased) frommilk at day #., respectively. Comparison betweenmilk exosomesat day - and day #. showed no DAP. The DAP identi,ed were related to biological functions such as uptake of metabolites,regulation of hemostasis and cellular development.Summary/Conclusion:Colostral exosomes have di3erent proteome pro,les than exosomes frommaturemilk, with signi,cancenot only for the physiological understanding of EV’s impact on the interaction between mother and newborn, but also for theprospective use of supplements with high economic potential such as milk and colostrum replacers.PS!&.&! Comparison of extracellular vesicles and proteins as a source of small RNA biomarkers incanine urineJenni Karttunen, PhD, Department of Veterinary Medicine, University of CambridgeLajos Kalmar, PhD, Department of Veterinary Medicine, University of Cambridge, UKSarah E. Stewart, Department of Biochemistry and Genetics, La Trobe Institute for Molecular Science, La Trobe University,Melbourne, VIC 0"40, AustraliaAndrew Grant, PhD, Department of Veterinary Medicine, University of Cambridge, UKFiona Karet Frankl, PhD, FRCP, FMedSci, Department of Medical Genetics, University of Cambridge, UKTimWilliams, PhD, Department of Veterinary Medicine, University of Cambridge, UKIntroduction:Urinary extracellular vesicles (EVs) and their RNA cargo are a novel source of biomarkers, however non-vesicularRNA is also present within urine. Here, we compared the small RNA pro,les of the EV and protein fractions of canine urine, todetermine their potential as a source of small RNA biomarkers.Methods: EV and protein fractions were obtained by size-exclusion chromatography (SEC) of ".!!µm ,ltered urine from,ve healthy control (HC) dogs and ,ve dogs with urinary tract infections (UTI). RNA from both fractions was analysedusing the Agilent Bioanalyzer small RNA chip and small RNA sequencing. The raw sequences were compared to examine thefraction-speci,c pro,les. EV and protein fractions fromHC and UTI samples were compared to identify di3erentially expressedsequences.Results: In HC samples, the protein fractions included more small RNA compared to EV fractions (0"..-0"-.! vs !.2-!".0 ng/mlof starting urine, respectively). After sequencing, ##"""–!"""" and #-"""–-0""" di3erent raw sequences (>5 copies) were iden-ti,ed from the HC protein and EV fractions respectively, -""–5""" of which were shared. When UTI samples were compared toHC, no sequences were di3erentially expressed in the EV fractions (adjusted p-value< "."5) and #-!were di3erentially expressedin protein fractions. Principle component analysis separated the UTI and HC groups based on the small RNA pro,les of the pro-tein fractions, but not based on the small RNA pro,les of EV fractions.Summary/Conclusion: EV and protein fractions obtained after SEC of ".!! um ,ltered dog urine have distinct small RNApro,les. When HC and UTI samples were compared, di3erentially expressed RNA sequences were identi,ed in the proteinfractions, but no signi,cant di3erences in the EV fractions were demonstrated. These results suggest that UTI does not lead tochanges in the small RNA pro,le of small EVs in urine.

Comparative proteome profiling in exosomes derived from porcine colostrum versus mature milk

https://air.unimi.it/bitstream/2434/887805/2/Agazzi%20et%20al.%20ISEV%20%202021%20A.pdf

Comparative proteome profiling in exosomes derived from porcine colostrum versus mature milk

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