| 1. |
- Eldh, Maria, et al.
(författare)
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Proteomic Profiling of Tissue Exosomes Indicates Continuous Release of Malignant Exosomes in Urinary Bladder Cancer Patients, Even with Pathologically Undetectable Tumour
- 2021
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Ingår i: Cancers. - : MDPI. - 2072-6694. ; 13:13
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Tidskriftsartikel (refereegranskat)abstract
- Simple Summary Urinary bladder cancer (UBC) has a high recurrence rate, and biomarkers for different treatment strategies are highly needed. This study investigated the release of nanovesicles called exosomes from urinary bladder tissue from tumour-proximal sites as well as tumour-distant sites in transurethrally resected (TUR-B) patients with or without preoperative neoadjuvant chemotherapy prior to ensuing radical cystectomy-all without remaining visible tumour after TUR-B. We show that cancer-promoting exosomes were detected from both sites, suggesting that the previous tumour has altered the whole bladder tissue into a cancer-supporting milieu. The exosomes may originate from remaining pathologically undetectable cancer cells or transformed epithelial cells, and the study supports the notion of exosomes as mediators of metastatic spread and as potential biomarkers. It also supports early and radical removal of the bladder in urinary bladder cancer patients. Invasive urothelial bladder cancer (UBC) has high recurrence rates even after radical cystectomy (RC). Exosomes are membrane-bound nanovesicles, which have been shown to contribute to carcinogenesis and metastasis. We previously showed that urinary exosomes display a malignant profile in UBC patients despite the absence of detectable tumour. Here, we investigated exosomes from sampling sites close to or distant from the former tumour, aiming to understand the effect of the tumour on the local milieu. Ten patients scheduled for cystectomy after transurethral bladder resection (TUR-B), without remaining detectable tumour, were included. Exosomes were isolated from tissue explants of both the previous tumour site and distant bladder tissue. Proteins were quantified by mass spectrometry in seven patients. Exosomes from the previous tumour site were enriched in inflammatory but not cancer-related pathways compared to distant tissue. However, the 69 most abundant proteins in tissue-derived exosomes regardless of site, 20 of which were also found in urinary exosomes from our previous study, were enriched for cancer-related metabolic pathways and associated with poor prognosis in an external mRNA dataset. The enrichment of cancer-related pathways in the most abundant proteins, regardless of sampling site, confirms our hypothesis that despite the absence of detectable tumour, the entire bladder releases exosomes that contribute to metastasis and highlights the need for early RC.
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| 2. |
- Hiltbrunner, Stefanie
(författare)
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Exosomes : immunomodulators in cancer and therapy
- 2016
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Doktorsavhandling (övrigt vetenskapligt/konstnärligt)abstract
- Exosomes are nano-sized membrane vesicles derived from the late endosomal compartment. They are capable of transferring proteins, lipids and RNA between cells. B cell and dendritic cell (DC)-derived exosomes express major histocompatibility complex (MHC) class I and II, as well as costimulatory molecules (CD80/86) and can initiate T cell responses. Several clinical trials have shown DC-derived exosome-based cancer immune therapy to be safe but limited in inducing antigen-specific T cells. In contrast, tumour cell-derived exosomes can express immune inhibitory molecules and play an important role in spreading oncogenic activity by carrying tumour antigens, inducing angiogenesis at distant sites and preparing tissues for metastasis. This thesis aimed at I) analysing how to enhance the immunogenicity of exosomes for therapy, II) investigating whether MHC complexes on exosomes are needed to induce an anti-tumour immune response, III) comparing microvesicles and exosomes side by side for their immunogenic capacity, IV) understanding the metastatic process induced by tumour-derived exosomes from bladder cancer patients and whether certain exosomal proteins can be used as markers for diagnosis and prognosis. Study I reveals that exosomes loaded with the NKT cell ligand alpha-galactosylceramide (αGC) and the model antigen ovalbumin (OVA) activate NKT cells, induce strong NK and γδ T cell innate immune responses, and induce OVA-specific T and B cell responses far better than only OVA-loaded exosomes. Exosomes loaded with αGC/OVA decreased tumour growth and increased median survival compared to exosomes loaded with OVA only or soluble αGC + OVA alone in a B16 melanoma model. This study demonstrates how to increase the immunogenicity of DC-derived exosomes for cancer treatment. Study II demonstrates that exosomal MHC class I is dispensable for the induction of antigen-specific T cell responses if whole OVA is present. We show that OVA-loaded DC-derived exosomes from MHCI-/- mice induce antigen-specific T cells to the same extent as wild type exosomes. Even exosomes with MHC class I and II mismatch induced tumour-infiltrating CD8+ T cells and increase survival in a B16 melanoma model. This study provides new opportunities for the design of allogeneic exosome-based vaccines and therapies. Study III compares microvesicles (MV) and exosomes from OVA-exposed DCs side by side for their capacity to induce OVA-specific immune responses in vivo. MV and exosomes express similar surface markers but only exosomes induced OVA-specific CD8+ T cells and OVA-specific IgG antibodies. In contrast, MV induced a higher number of plasma cells. Finally, we found that exosomes contain more OVA compared to MV. We conclude that exosomes from DCs are superior in inducing antigen-specific immune responses in vivo compared to MVs, while MVs might activate the immune system unspecifically. Study IV evaluates the proteomic profile of exosomes from tumour tissue explants and urine from urinary bladder cancer patients. We show that exosomes from malignant or benign tissue can be distinguished by the proteomic profile and are involved in platelet, metabolic and immune signalling networks. We show that, even if no tumour is left, exosomes can express a metastatic memory phenotype which might be involved in cancer progression. In summary, this thesis gives new insights into how to design vesicle-based cancer vaccines and provide new opportunities for the use of allogeneic DC-derived exosomes in patients. In addition, we demonstrate that exosomes isolated from the urine of urinary bladder cancer patients express specific markers for malignancy, which provides new possibilities for diagnostic strategies.
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| 3. |
- Hiltbrunner, Stefanie, et al.
(författare)
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Urinary Exosomes from Bladder Cancer Patients Show a Residual Cancer Phenotype despite Complete Pathological Downstaging
- 2020
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Ingår i: Scientific Reports. - : Nature Publishing Group. - 2045-2322. ; 10:1
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Tidskriftsartikel (refereegranskat)abstract
- Invasive urinary bladder cancer shows high recurrence rates after cystectomy even with apparent complete downstaging at cystectomy. Exosomes are nano-sized vesicles important in cell-cell communication, which have been hypothesized to contribute to cancer dissemination and recurrence. The aim of this study was to investigate if pro-carcinogenic exosomes could be detected in urine from histologically downstaged bladder cancer patients. 13 Patients were included in this study. Paired ureter and urine samples from nine patients underwent mass spectrometry, while samples from the remaining patients were used for exosome characterization. At cystectomy, exosomes were isolated from bladder and ureter urine, whereafter quantitative proteome profiling was performed. Urinary exosomes clustered based on whether they came from the bladder, with tumour contact, or the ureters, without tumour contact, even though all came from completely downstaged patients. Proteins overexpressed in exosomes derived from bladder urine contained several oncogenes and were mainly associated with tumour metabolism pathways. Although patients were histologically tumour-free at cystectomy, the bladder urine contained exosomes with a carcinogenic metabolic profile. This suggests a continuous release of exosomes from the bladder, which may promote recurrence at distant sites through metabolic rewiring, even after apparent complete downstaging. These exosomes, coming from either undetected cancer cells or partly transformed cells, are likely to increase the risk of metastasis and encourages cystectomy even in completely downstaged patients.
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| 4. |
- Levänen, Bettina, et al.
(författare)
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Altered microRNA profiles in bronchoalveolar lavage fluid exosomes in asthmatic patients.
- 2013
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Ingår i: Journal of Allergy and Clinical Immunology. - : Elsevier BV. - 0091-6749 .- 1097-6825. ; 131:3, s. 894-903
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Tidskriftsartikel (refereegranskat)abstract
- BACKGROUND: Asthma is characterized by increased airway narrowing in response to nonspecific stimuli. The disorder is influenced by both environmental and genetic factors. Exosomes are nanosized vesicles of endosomal origin released from inflammatory and epithelial cells that have been implicated in asthma. In this study we characterized the microRNA (miRNA) content of exosomes in healthy control subjects and patients with mild intermittent asthma both at unprovoked baseline and in response to environmental challenge.OBJECTIVE: To investigate alterations in bronchoalveolar lavage fluid (BALF) exosomal miRNA profiles due to asthma, and following subway air exposure.METHODS: Exosomes were isolated from BALF from healthy control subjects (n = 10) and patients with mild intermittent asthma (n = 10) after subway and control exposures. Exosomal RNA was analyzed by using microarrays containing probes for 894 human miRNAs, and selected findings were validated with quantitative RT-PCR. Results were analyzed by using multivariate modeling.RESULTS: The presence of miRNAs was confirmed in exosomes from BALF of both asthmatic patients and healthy control subjects. Significant differences in BALF exosomal miRNA was detected for 24 miRNAs with a subset of 16 miRNAs, including members of the let-7 and miRNA-200 families, providing robust classification of patients with mild nonsymptomatic asthma from healthy subjects with 72% cross-validated predictive power (Q(2) = 0.72). In contrast, subway exposure did not cause any significant alterations in miRNA profiles.CONCLUSION: These studies demonstrate substantial differences in exosomal miRNA profiles between healthy subjects and patients with unprovoked, mild, stable asthma. These changes might be important in the inflammatory response leading to bronchial hyperresponsiveness and asthma.
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