Browsing by Subject "Exosome"
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Item Open Access Biological properties of extracellular vesicles and their physiological functions(Taylor & Francis, 2015) Yáñez-Mó, M.; Siljander, P. R. M.; Andreu, Z.; Zavec, A. B.; Borràs, F. E.; Buzas, E. I.; Buzas, K.; Casal, E.; Cappello, F.; Carvalho, J.; Colás, E.; Cordeiro-Da, S. A.; Fais, S.; Falcon-Perez, J. M.; Ghobrial, I. M.; Giebel, B.; Gimona, M.; Graner, M.; Gursel, I.; Gursel, M.; Heegaard, N. H. H.; Hendrix, A.; Kierulf, P.; Kokubun, K.; Kosanovic, M.; Kralj-Iglic, V.; Krämer-Albers, E. M.; Laitinen, S.; Lässer, C.; Lener, T.; Ligeti, E.; Line, A.; Lipps, G.; Llorente, A.; Lötvall, J.; Manček-Keber, M.; Marcilla, A.; Mittelbrunn, M.; Nazarenko, I.; Nolte-'t Hoen, E. N. M.; Nyman, T. A.; O'Driscoll, L.; Olivan, M.; Oliveira, C.; Pállinger, E.; Del Portillo, H. A.; Reventós, J.; Rigau, M.; Rohde, E.; Sammar, M.; Sánchez-Madrid, F.; Santarém, N.; Schallmoser, K.; Ostenfeld, M. S.; Stoorvogel, W.; Stukelj, R.; Grein V. D. S.G.; Helena,ü V. M.; Wauben, M. H. M.; De Wever, O.In the past decade, extracellular vesicles (EVs) have been recognized as potent vehicles of intercellular communication, both in prokaryotes and eukaryotes. This is due to their capacity to transfer proteins, lipids and nucleic acids, thereby influencing various physiological and pathological functions of both recipient and parent cells.While intensive investigation has targeted the role of EVs in different pathological processes, for example, in cancer and autoimmune diseases, the EV-mediated maintenance of homeostasis and the regulation of physiological functions have remained less explored. Here, we provide a comprehensive overview of the current understanding of the physiological roles of EVs, which has been written by crowd-sourcing, drawing on the unique EV expertise of academia-based scientists, clinicians and industry based in 27 European countries, the United States and Australia. This review is intended to be of relevance to both researchers already working on EV biology and to newcomers who will encounter this universal cell biological system. Therefore, here we address the molecular contents and functions of EVs in various tissues and body fluids from cell systems to organs. We also review the physiological mechanisms of EVs in bacteria, lower eukaryotes and plants to highlight the functional uniformity of this emerging communication system.Item Open Access Circulating LL37 targets plasma extracellular vesicles to immune cells and intensifies Behçet's disease severity(Taylor and Francis, 2017-02) Kahraman, T.; Gucluler, G.; Simsek, I.; Yagci, F. C.; Yildirim, M.; Ozen, C.; Dinc, A.; Gursel, M.; Ikromzoda, L.; Sutlu, T.; Gay, S.; Gursel, I.Behçet's disease (BD) activity is characterised by sustained, over-exuberant immune activation, yet the underlying mechanisms leading to active BD state are poorly defined. Herein, we show that the human cathelicidin derived antimicrobial peptide LL37 associates with and directs plasma extracellular vesicles (EV) to immune cells, thereby leading to enhanced immune activation aggravating BD pathology. Notably, disease activity was correlated with elevated levels of circulating LL37 and EV plasma concentration. Stimulation of healthy PBMC with active BD patient EVs induced heightened IL1β, IFNα, IL6 and IP10 secretion compared to healthy and inactive BD EVs. Remarkably, when mixed with LL37, healthy plasma-EVs triggered a robust immune activation replicating the pathology inducing properties of BD EVs. The findings of this study could be of clinical interest in the management of BD, implicating LL37/EV association as one of the major contributors of BD pathogenesis.Item Open Access CpG ODN loaded exosome nanovesicles: enhanced immunostimulatory activity(Turkish Society of Immunology, 2012-04) Güçlüler, Gözde; Kahraman, Tamer; Gürsel, İhsanExosomes are naturally occurring, membranous nanovesicles known to function as intercellular communication vectors. To explore whether these naturally occuring bilayer vesicles could be exploited as a nucleic acid delivery system we intentionally loaded exosomes with TLR9 ligands and studied their immunostimulatory properties. METHODS: Exosomes were isolated from RAW264.7 and EG-7 cell supernatants by differential centrifugation, filtration and ultracentrifugation and were loaded with CpG ODNs via dehydration-rehydration protocol. Splenocytes were stimulated and analyzed by flow cytometry for cell surface marker upregulation, intracellular cytokine production and co-stimulatory molecule upregulation. Confocal microscopy analyses were done to assess internalization properties of these nanovesciles. Cell supernatants were studied by ELISA to assess Th1 biased cytokine/chemokine secretion.Item Open Access Evidence-Based Clinical Use of Nanoscale Extracellular Vesicles in Nanomedicine(American Chemical Society, 2016-03) Fais, S.; O'Driscoll, L.; Borras, F. E.; Buzas, E.; Camussi, G.; Cappello, F.; Carvalho, J.; Cordeiro Da Silva, A.; Del Portillo, H.; El Andaloussi, S.; Ficko Trček, T.; Furlan, R.; Hendrix, A.; Gursel, I.; Kralj-Iglic, V.; Kaeffer, B.; Kosanovic, M.; Lekka, M. E.; Lipps, G.; Logozzi, M.; Marcilla, A.; Sammar, M.; Llorente, A.; Nazarenko, I.; Oliveira, C.; Pocsfalvi, G.; Rajendran, L.; Raposo, G.; Rohde, E.; Siljander, P.; Van, N. G.; Vasconcelos, M. H.; Yáñez-Mó, M.; Yliperttula, M. L.; Zarovni, N.; Zavec, A. B.; Giebel, B.Recent research has demonstrated that all body fluids assessed contain substantial amounts of vesicles that range in size from 30 to 1000 nm and that are surrounded by phospholipid membranes containing different membrane microdomains such as lipid rafts and caveolae. The most prominent representatives of these so-called extracellular vesicles (EVs) are nanosized exosomes (70-150 nm), which are derivatives of the endosomal system, and microvesicles (100-1000 nm), which are produced by outward budding of the plasma membrane. Nanosized EVs are released by almost all cell types and mediate targeted intercellular communication under physiological and pathophysiological conditions. Containing cell-type-specific signatures, EVs have been proposed as biomarkers in a variety of diseases. Furthermore, according to their physical functions, EVs of selected cell types have been used as therapeutic agents in immune therapy, vaccination trials, regenerative medicine, and drug delivery. Undoubtedly, the rapidly emerging field of basic and applied EV research will significantly influence the biomedicinal landscape in the future. In this Perspective, we, a network of European scientists from clinical, academic, and industry settings collaborating through the H2020 European Cooperation in Science and Technology (COST) program European Network on Microvesicles and Exosomes in Health and Disease (ME-HAD), demonstrate the high potential of nanosized EVs for both diagnostic and therapeutic (i.e., theranostic) areas of nanomedicine.Item Open Access In vitro and in vivo immunomodulatory effects of extracellular vesicles(Bilkent University, 2013) Şahin, MehmetThe major theme of this thesis was to characterize and understand the immunomodulatory potential of extracellular vesicles (EVs) isolated from different mouse and human cell lines on immune cells. To this end, we first purified and characterized the particle nature of exosomes and microparticles (MPs) and exosomes by AFM and DLS analyses. Next, we documented their in vitro or in vivo differential uptake/internalization kinetics by immune cells. Finally, we tested the potential application of EVs as a drug delivery system. The physicochemical characterization studies confirmed that EVs were in vesicular form and upon reconstitution followed by lyophilization they retained their original sizes (i.e. size ranges were 100-150 nm for exosomes, and 250-500 nm for MPs, respectively). MPs derived from mouse cell lines (macrophage, T-cell and fibroblast) were stained and administered in vitro and in vivo to track their internalization by immune cells. When incubated in culture, origins of MPs greatly affected uptake rate and ratio by different immune cells. This differential internalization pattern of distinct MPs was reproduced when they were injected i.p. to mice. When incubated in culture B-cells took up the most MPs from fibroblast origin cells. Macrophages of peritoneal exudate cells took up RAW264.7 derived MPs at the highest level upon 24h post-ip injection. We tested whether MPs have a role in transportation or presentation of bacterial products during an ongoing infection by adhering circulating pathogenic by-products and carrying them to distant immune cells. Our data suggest that MPs could contribute to the severity of ongoing infection in vivo, since MPs from different cells can adhere distinct ligands such as LPS or DNA or even RNA and transmit these ligands to naive innate immune cells augmenting the immunostimulatory response raised against these ligands. Lastly, the immunotherapeutic potential of EVs harboring immunosuppressive synthetic oligodeoxynucleotide sequence namely, A151 was tested following dehydrationrehydration method on mouse splenocytes. EV-associated A151 displayed improved inhibition of immune activation triggered by TLR7/8 and TLR9 ligands on spleen cells. When taken together, this study established that various types of EVs derived from different cells induces plethora of activities on immune cells. Furthermore, EVs are potential drug carrier systems when loaded externally with suitable agents and can be harnessed in immunotherapy of diseases.Item Open Access Label-free identification of exosomes using raman spectroscopy and machine learning(Wiley-VCH Verlag GmbH & Co. KGaA, 2023-01-15) Parlatan, U.; Ozen, M.O.; Kecoglu, I.; Koyuncu, B.; Torun, H.; Khalafkhany, D.; Loc, I.; Ogut, M.G.; Inci, Fatih; Akin, D.; Solaroglu, I.; Ozoren, N.; Unlu, M. B.; Demirci, U.Exosomes, nano-sized extracellular vesicles (EVs) secreted from cells, carry various cargo molecules reflecting their cells of origin. As EV content, structure, and size are highly heterogeneous, their classification via cargo molecules by determining their origin is challenging. Here, a method is presented combining surface-enhanced Raman spectroscopy (SERS) with machine learning algorithms to employ the classification of EVs derived from five different cell lines to reveal their cellular origins. Using an artificial neural network algorithm, it is shown that the label-free Raman spectroscopy method's prediction ratio correlates with the ratio of HT-1080 exosomes in the mixture. This machine learning-assisted SERS method enables a new direction through label-free investigation of EV preparations by differentiating cancer cell-derived exosomes from those of healthy. This approach will potentially open up new avenues of research for early detection and monitoring of various diseases, including cancer.Item Open Access Mirna based identification of prostate cancer by investigation of urinary exosomes(Bilkent University, 2020-08) Bozbeyoğlu, NazProstate cancer is one of the most incident cancer subtypes with high mortality rate. Currently, diagnosis of prostate cancer is based on rectal examination, Prostate Specific Antigen (PSA) testing and biopsy. Normal range of PSA is defined as 0-4 ng/ml and individuals with higher PSA levels are considered as potential prostate cancer patients. However, PSA fluctuates as a result of many factors and it is shown to increase with age. Thus, PSA testing causes significantly high false positive results and many healthy men have biopsy unnecessarily due to high PSA levels or they even get overtreated. This situation has huge psychological as well as financial effects on these people. Herein, we investigated the diagnostic potential of urinary exosomal microRNAs (miRNA) in prostate cancer. Rather than investigation of cellular miRNAs, we focused on exosomal miRNAs because of high integrity of exosomes and their abundance in many biofluids including urine. Development of a sensitive diagnostic method from urine would be advantageous because accurate diagnosis of prostate cancer would be possible by a non-invasive procedure. miRNAs are the small non-coding RNAs and they suppress expression of target genes via degradation of mRNA or post-translational regulation. miRNAs have high potential as cancer biomarkers because they can act as tumor suppressor and repress oncogenic gene expression or function as oncogenic miRNA and suppress tumor suppressor gene expression. For this reason, we identified several candidate exosomal tumor suppressor and oncogenic miRNAs and continued our study with the most potent ones. At the beginning of the study, we validated that we efficiently isolated exosomes via several techniques such as flow cytometry, Dynamic Light Scattering (DLS), Nanoparticle Tracking Analysis (NTA) and Transmission Electron Microscopy (TEM). Then, we studied differential expression of candidate miRNAs in prostate cancer PC-3 cell line. Similar to our literature search findings, we found that expression levels of miR-107, miR-139, miR-145 and miR-204 were significantly lower in PC-3 exosomes in comparison to healthy urinary exosomes. On the other hand, oncomiRs; miR-21-5p, miR-375-5p and miR-574 3p were upregulated in PC-3 cell line exosomes. Of note, expression of another candidate miRNA; miR-30a was almost the same in PC-3 exosomes with healthy controls. We used these results as preliminary data and collected urine specimens from 17 prostate cancer patients. We firstly analyzed their PSA level-age and PSA Level- Gleason Score correlations. Our analyses revealed that PSA level was increasing with age and PSA was not correlated with Gleason Score. We concluded that PSA was being affected by several reasons in addition to tumor formation and it was not correlated with disease progression. Again, this was a finding which supported that a more sensitive diagnosis method than PSA testing was necessary for prostate cancer. When we detected expression levels of candidate miRNAs in urinary exosomes of prostate cancer patients and healthy controls, we observed that miR-107, miR-139, miR-145 and miR-204 were downregulated whereas miR-375-5p was upregulated in patients’ urinary exosomes. For miR-21, there was a very slight upregulation and miR-30a-5p and miR-574-3p levels were almost the same with healthy controls. Further, we wondered the diagnostic potential of these miRNAs in our patient cohort and we performed Receiver Operator Characteristic (ROC) Curve analysis for them. When they were used as combination, our miRNAs had 77% (AUC=0.7731) accuracy in distinguishing patients from healthy controls. After that, we examined miRNA dysregulations in patients with different PSA levels with the hypothesis that they may have different miRNA expression profiles. We saw that expression of miRNAs in exosomes patients with PSA<10 ng/ml and PSA=10-15 ng/ml were very similar with our expectations; downregulation of tumor suppressor and upregulation in oncogenic miRNAs whereas patients with PSA>15 ng/ml had a unique profile and all miRNAs were downregulated. Due to sample size limitations, we only tested diagnostic potential of candidate miRNAs in exosomes of patients with PSA<10 ng/ml and observed that AUC was 0.8500 so we could discriminate patients and healthy controls with 85% accuracy by using our candidate miRNA panel in testing. Taken together, our findings indicated that candidate urinary exosomal tumor suppressor and oncogenic miRNAs which we suggested in thesis are very potent in diagnosis of prostate cancer with differential expressions in prostate cancer patients with different PSA levels and this study opens the way for development of a noninvasive prostate cancer diagnosis method.Item Open Access The effect of exosomes released from apheresis platelet concentrates under the impact of gamma irradiation and storage time upon platelet aggregation and hemostasis(Edizioni SIMTI, 2023) Bal, Salih H.; Sağdilek, E.; Karaçay, M.; Kızmaz, Muhammed A.; Kumaş, Levent T.; Can, Fatma E.; Yıldırım, M.; Canavar-Yıldırım, Tuğçe; Koşay-Gülkaya, D.; Yöyen-Ermiş, D.; Budak, F.; Heper, Y.