Advances in membranous vesicle and exosome proteomics improving biological understanding and biomarker discovery

Raimondo, Francesca; Morosi, Lavinia; Chinello, Clizia; Magni, Fulvio; Pitto, Marina

doi:10.1002/pmic.201000422

Cited by 283 publications

(251 citation statements)

References 94 publications

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“…2 and 3) agree with data present in the literature. 30 Moreover, there are no important morphological differences between exosomes in the two groups (patients and controls), as shown also by the morphometric analysis ( Fig. 2A, B and 3).…”

Section: Urinary Vesicle Isolation and Characterizationsupporting

confidence: 51%

“…7B), and we assessed the presence of many typical exosomal proteins, such as the component of the ESCRT machinery (TSG101), proteins involved in trafficking View Article Online and membrane fusion (annexins), and tetraspanins such as CD9, possibly correlated to the exosome biogenesis mechanism. 30 Moreover, many other functional classes were recognized, such as metabolic enzymes (i.e. triosophosphate isomerase, isocitrate dehydrogenase), proteins involved in signalling (i.e.…”

Section: Protein Identificationmentioning

confidence: 99%

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Differential protein profiling of renal cell carcinoma urinary exosomes

et al. 2013

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aRenal cell carcinoma (RCC) accounts for about 3% of all human malignancies and its incidence is increasing. There are no standard biomarkers currently used in the clinical management of patients with renal cell carcinoma. A promising strategy for new biomarker detection is comparative proteomics of urinary exosomes (UE), nanovesicles released by every epithelial cell facing the urinary space, enriched in renal proteins and excluding high-abundance plasmatic proteins, such as albumin. Aim of the work is to establish the protein profile of exosomes isolated from urines of RCC patient compared with control subjects. We enrolled 29 clear cell RCC patients and 23 control healthy subjects (CTRL), age and sex-matched, for urine collection and vesicle isolation by differential centrifugation. Such vesicles were morphologically and biochemically characterized and proved to share exosome properties.Proteomic analysis, performed on 9 urinary exosome (UE) pooled samples by gel based digestion followed by LC-MS/MS, led to the identification of 261 proteins from CTRL subject UE and 186 from RCC patient UE, and demonstrated that most of the identified proteins are membrane associated or cytoplasmic. Moreover, about a half of identified proteins are not shared between RCC and control UE.Starting from these observations, and from the literature, we selected a panel of 10 proteins, whose UE differential content was subjected to immunoblotting validation. Results show for the first time that RCC UE protein content is substantially and reproducibly different from control UE, and that these differences may provide clues for new RCC biomarker discovery.

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Section: Urinary Vesicle Isolation and Characterizationsupporting

confidence: 51%

Section: Protein Identificationmentioning

confidence: 99%

Differential protein profiling of renal cell carcinoma urinary exosomes

et al. 2013

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“…Among them are CD9, CD63, CD81, and CD82, which are often used as exosome markers. 30,113 Some tetraspanins are selectively enriched in exosomes compared with their parent cells. An example is CD9, which was found to be more than 10-fold enriched in dendritic cell-derived exosomes.…”

Section: Tetraspaninsmentioning

confidence: 99%

“…124 Thus, the fusogenic properties of tetraspanins appear to be dependent on cell type. Interestingly, the tetraspanins which are often found to be enriched in exosomes (CD9, CD63, CD81, and CD82) 30,113 have been shown to be involved in the migration of dendritic cells. In a chemotaxis assay, antibodies against the single tetraspanins increased migration of these cells by 50%-70%, while 100% increased migration was observed when these antibodies were combined.…”

Section: Tetraspaninsmentioning

confidence: 99%

Exosome mimetics: a novel class of drug delivery systems

Kooijmans¹,

Vader²,

Dommelen³

et al. 2012

IJN

251

124

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Abstract:The identification of extracellular phospholipid vesicles as conveyors of cellular information has created excitement in the field of drug delivery. Biological therapeutics, including short interfering RNA and recombinant proteins, are prone to degradation, have limited ability to cross biological membranes, and may elicit immune responses. Therefore, delivery systems for such drugs are under intensive investigation. Exploiting extracellular vesicles as carriers for biological therapeutics is a promising strategy to overcome these issues and to achieve efficient delivery to the cytosol of target cells. Exosomes are a well studied class of extracellular vesicles known to carry proteins and nucleic acids, making them especially suitable for such strategies. However, the considerable complexity and the related high chance of off-target effects of these carriers are major barriers for translation to the clinic. Given that it is well possible that not all components of exosomes are required for their proper functioning, an alternative strategy would be to mimic these vesicles synthetically. By assembly of liposomes harboring only crucial components of natural exosomes, functional exosome mimetics may be created. The low complexity and use of well characterized components strongly increase the pharmaceutical acceptability of such systems. However, exosomal components that would be required for the assembly of functional exosome mimetics remain to be identified. This review provides insights into the composition and functional properties of exosomes, and focuses on components which could be used to enhance the drug delivery properties of exosome mimetics.

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“…This chapter will review current knowledge pertaining to exosomes, describe possible uses of exosomes in immunotherapy, and address challenges and future directions in bringing exosome-based cancer vaccines or immunotherapies closer to clinical reality. amount of publications devoted to the subject just within the past decade (Raimondo et al, 2011). Many types of vesicles have been described in the literature having quite heterogeneous size, protein content, RNA content and origin.…”

Section: Introductionmentioning

confidence: 99%