Abstracts from the Third International Meeting of ISEV 2. Rotterdam, The Netherlands, April 3. May 3rd, 2. 01. 4J Extracell Vesicles.
Copyright © 2. 01. Journal of Extracellular Vesicles. This is an Open Access article distributed under the terms of the Creative Commons Attribution- Noncommercial 3. Unported License, permitting all non- commercial use, distribution, and reproduction in any medium, provided the original work is properly cited. Registration 8: 3. Welcome & Networking coffee Arcadis Room 8: 3.
Arcadis Room. Setting up posters (Poster Sessions 1. A, 1. B, 1. C, 3. A, 3. C, 2. B, 1. A) 8: 3. 0- 9: 0. Sponsor Exhibition Jurriaans Foyer/Mandele room 1. Opening Session Willem Burger room 1.
Questions and Answers from the Community. It doesn't. The page that you see when you ask a new question is the page that everyone will see. SAM.gov The System for Award Management (SAM) is the Official U.S. Government system that consolidated the capabilities of CCR/FedReg, ORCA, and EPLS. Plan1 Capability List 2524624-6D 2524634-1 2524634-8 2524638B 2524640-1 2524640-8 2524645B 2524648B 2524650B 2524654-21 2524665 2524665-1 2524673-8 2524681-1.
Welcome/Development ISEV 2. Jan Lötvall, President of ISEV ISEV 2.
· NHLBI 081. Passive MRI. it is sufficient to state for purposes of a Justification and Approval pursuant to FAR 6.302-5 that the project is a SBIR.
Rotterdam 1. Marca Wauben, President LOCWillem Burger room. Symposium Session 1. A - EV in the tumor micro- environment. Chair: Antonella Bongiovanni and Janusz Rak 1.
O1. A- 0. 01. Intravital imaging of extracellular vesicle exchange between tumour cells. Anoek Zomer and Jacco Van Rheenen. Cancer Biophysics, Hubrecht Institute, Utrecht, The Netherlands Introduction: Most cancer cells release heterogeneous populations of extracellular vesicles containing proteins, lipids and nucleic acids that reflect, at least in part, the cell of origin. Accumulating data obtained in vitro has demonstrated that the molecules packaged in extracellular vesicles can be functionally transferred into a variety of recipient cells affecting their gene expression and behaviour. Vesicles released by human tumour cells have been shown to interfere with anti- tumor immunity, and it has been suggested that tumor- released vesicles play a role in autocrine signalling, the formation of a pre- metastatic niche, drug resistance, angiogenesis and stromal remodelling. However, how tumour cell behaviour is altered upon extracellular vesicle uptake in vivo is largely unknown.
Methods: We developed a novel method to visualize the functional transfer of molecules packaged in extracellular vesicles in real- time in living mice, providing further opportunities to study in detail the distribution and biological relevance of extracellular vesicles between various cells and tissues. Results: Using this method, we show in living mice that local and systemic transfer of molecules carried by extracellular vesicles is a physiological process that is directly coupled to the migratory behaviour and metastatic capacity of recipient breast cancer cells. Summary/conclusion: The data presented here are consistent with the idea that tumour cells do not act autonomously, but can share proteins and nucleic acids with other tumour cells, locally and systemically. These results shed light on the mutual influence of cancer cells and draw a new perspective on the complexity of intercellular communication in diseases such as cancer. O1. A- 0. 02. High- resolution imaging of uptake and processing of prostate cancer- derived exosomes in living prostate epithelial cells. Thomas Hartjes. 1, Diederick Duijvesz.
Johan Slotman. 3, Adriaan Houtsmuller. Guido Jenster. 2 and Martin van Royen. Department of Pathology, Erasmus MC, Rotterdam, The Netherlands; 2. Department of Urology, Erasmus MC, Rotterdam, The Netherlands; 3. Erasmus Optical Imaging Centre, Erasmus MC, Rotterdam, The Netherlands Introduction: Prostate cancer (PCa) is the most common malignancy in men.
Whereas overall survival of patients with early- diagnosed localized PCa is high, metastasized PCa decreases survival dramatically. Tumor cells influence their microenvironment to enhance tumour progression and metastasis. Recently it was found that tumor- derived exosomes play a role in this communication between tumour cells and surrounding stromal and epithelial cells. We aim to provide novel insight into the exosome- mediated mechanisms by which PCa cells influence their microenvironment. Using live cell confocal imaging and high- resolution microscopy, the different stages of uptake and intracellular processing of PCa- derived exosomes by prostate epithelial cells were visualized.
This will be accompanied by development of approaches to block exosome function. Methods: Exosomes from PCa cell line DU1. PKH2. 6/6. 7). Uptake and further processing of fluorescently labelled exosomes by non- tumorigenic prostate epithelial cells are followed on different time scales, from seconds to multiple hours, by live cell imaging using conventional confocal microscopy and, for high- speed imaging, spinning disk microscopy. Different stages of exosome uptake and co- localization of exosomes with specific proteins are studied in more detail using structured illumination super- resolution microscopy (SIM). Results: Confocal time- lapse images show a rapidly initiated and continuous uptake of individual fluorescently labelled exosomes by living PNT2.
C2 cells. High- speed spinning disk microscopy shows that internalized exosomes are transported via microtubules. This was extended with super- resolution co- localization studies between exosomes and proteins involved in endocytosis that showed processing of internalized exosomes through endosome and lysosome pathways.
Summary/conclusion: Different imaging approaches enabled us to visualize subsequent steps and dynamics of exosome interaction, uptake and further processing by target cells. Using high- speed imaging and super- resolution SIM allows us to further unravel the molecular mechanisms of action and the role of exosomes in PCa. O1. A- 0. 03. Hodgkin lymphoma cells dispatch shedding- sensitive signalling proteins on microvesicles to transiently interact with the tumour microenvironment. Hinrich P. Hansen. Adriana F. Paes Leme. Maria Dams. 1 and Elke Pogge von Strandmann.
University of Cologne, Cologne, Germany; 2. Brazilian Biosciences National Laboratory, LNBio, CNPEM, Campinas, Brazil Introduction: Hodgkin lymphoma (HL)- affected lymphoid tissue contains only a few disseminated tumour cells, which stimulate immune cells in the tumour microenvironment not to suppress but to support tumour growth. Extracellular vesicles (EVs) from the tumour cells participate in this communication (Hansen et al., 2. J Pathol). Methods: EVs were isolated by sequential ultracentrifugation and investigated by electron and confocal microscopy as well as bead- coupled flow cytometry. Tryptic peptides of EV proteins were analysed by LTQ Velos Orbitrap mass spectrometry. Immune cell migration was performed by chemotaxis experiments and video time- lapse microscopy.
Results: Mass spectrometry of EVs from 4 different cell lines showed a strong expression of the sheddases ADAM1. ADAM1. 7, and among their potential substrates were many functional molecules, including CD3.
CD4. 4, CD1. 66/ALCAM, Notch. Eighteen- hour incubation of isolated EVs reduced the CD3. CD4. 4 to 4. 1. 3% versus metalloproteinase- inhibited aliquots (BB- 9. M), whereas the expression of CD4. CD7. 0 (9. 0. 6%) remained rather uninfluenced. CD3. 0 is a HL- selective receptor of the tumour necrosis factor receptor family (TNFRSF8).
We showed that EVs expressing CD3. CD3. 0- dependent manner.
The shedding product (s. CD3. 0) lacked this function but itself served as a chemokine for polymorphonuclear leukocytes. Summary/conclusion: Metalloproteinases alter the functionality of EVs through selective cleavage of certain membrane proteins. As an example, the soluble ectodomain of CD3. CD3. 0) and CD3. 0+ EVs showed different functionality, suggesting that metalloproteinases, by converting selected functional proteins, are able to coordinate a time- limited functionality of EVs in tumour tissue. O1. A- 0. 04. Role of nasopharyngeal carcinoma- derived exosomes on the development of tolerogenic immature dendritic cells.
Dhafer Mrizak. 1, Joshua Mason. Zachary Fitzpatrick. Yvan De Launoit. 2, Pierre Busson. Olivier Morales. 2 and Nadira Delhem. Institut de Biologie de Lille/CNRS, Lille, France; 2. UMR 8. 16. 1, Institut de Biologie de Lille/CNRS, Lille, France; 3. UMR 8. 12. 6, Institut Gustave Roussy, Villejuif, France Introduction: We have shown previously that nasopharyngeal carcinoma (NPC) immune escape is closely related to the presence of regulatory T cells (Treg) recruited by NPC- derived exosomes.
We investigate here the role of these tumour exosomes on dendritic cells (DCs), which are highly represented in the immature state surrounding the tumour and that are able to induce the differentiation of naive CD4 T cells into Treg.