Am J Cancer Res 2011;1(1):98-110 www.ajcr.us /ISSN:2156-6976/ajcr0000008
Review Article
Exosome/microvesicle-mediated epigenetic reprogramming of cells
Giovanni Camussi1, Maria-Chiara Deregibus1, Stefania Bruno2, Cristina Grange1, Valentina Fonsato1, Ciro Tetta3
1Department of Internal Medicine, Centre for Molecular Biotechnology and Centre for Research in Experimental Medicine (CeRMS), Torino, Italy and 2Sis-Ter, Palazzo Pignano, Italy and 3Fresenius Medical Care, Bad Homburg, Germany.
Received October 19, 2010; accepted October 21, 2010; Epub October 22, 2010; Published January 1, 2011
Abstract: Microvesicles (MVs) are released by different cell types and may remain in the extracellular space in prox- imity of the cell of origin or may enter the biological fluids. MVs released by tumor cells are detectable in patients with cancer and their number in the circulation correlates with poor prognosis. Recent studies demonstrated that MVs may act as mediator of cell-to-cell communication thus ensuring short- and long-range exchange of information. Due to their pleyotropic effects, MVs may play a role in the prothrombotic state associated with cancer as well as in can- cer development and progression. It has been recently shown that MVs may induce epigenetic changes in target cells by transferring genetic information. This finding suggests that tumor and stromal cells may talk each other via MVs to establish a favorable tumor niche and to promote tumor growth, invasiveness and progression. Moreover, MVs con- tain genetic material under the form of mRNA and microRNA, that may allow an easy screening for cancer genetic markers and offer new diagnostic and prognostic information. This review presents an overview of the many biologi- cal actions of MVs and of the potential role of MV-mediated exchange of genetic information among cells in tumor biology.
Keywords: Microvesicles, exosomes, angiogenesis, tumor niche, stem cells
Introduction
Exchange of information between cells may in- volve soluble factors or direct cell-to-cell con- tact. This may include cytonemes and tunneling nanotubules. Cytonemes connect neighboring cells enabling ligand-receptor-mediated transfer of surface-associated molecules. Tunneling nanotubules, by establishing conduits between cells, allow the transfer not only of surface molecules but also of cytoplasmic components [1, 2]. In addition, cells may communicate through membrane transfer by the secretion of exosomes/microvesicles [3]. Such membrane transfer is associated with the acquisition of functional properties in the recipient cells re- lated to the transferred proteins, receptors and/ or bioactive lipids. Recent studies indicate that exosomes/microvesicles shuttle mRNAs and microRNAs, raising the possibility that the trans-
fer of genetic information might alter the func- tion of recipient cells. The vesicles detectable both in vitro and in vivo are a mixed population of exosomes derived from the endosomal mem- brane compartment [4, 5] and of shedding vesi- cles, originated by direct budding from the cell plasma membrane [6]. For this reason in the present review we will call them collectively mi- crovesicles (MVs). Once secreted, MVs may re- main in the extracellular space in proximity of the cell of origin or may enter the biological flu- ids such as plasma, urine, milk, cerebrospinal fluid, amniotic fluid and tumor effusions, thus allowing long-range exchange of MV-mediated information. In normal subjects, the majority of MVs present in the circulation, designed also as microparticles, are derived from platelets [7], and in a smaller amount from other blood cells and endothelial cells [8]. However, many cell types including tumor cells are able to release

MVs and in cancer patients tumor-derived MVs are detectable within the biological fluids [9, 10].
Intracellular origin and characteristics of re- leased MVs
Irrespectively from their origin, MVs are circular membrane fragments retaining the characteris- tics of the cell of origin and containing cytosol. Depending on their intracellular origin and the mechanisms of formation, MVs may be distin- guished in shedding vesicles or exosomes.
Shedding vesicles, also named as ectosomes, microparticles or exovesicles, are rather hetero- geneous with a size ranging from 100 nm to 1 μm. They are formed by budding of the cell membrane producing small cytoplasmic protru- sions that undergo detachment from the cell surface (Figure 1A). This process depends on calcium influx, cytoskeleton reorganization and curvature-mediated lateral redistribution of membrane components with the formation of membrane nanodomains [11]. Shedding vesi- cles expose on their surface large amounts of phosphatydilserine and are enriched in proteins associated with membrane lipid rafts [12]. Their formation involves an increase of calcium ions that inhibits translocase and induces activation of scramblase that translocates phosphatydil- serine from the inner leaflet of the cell mem- brane bilayer to the outer [13]. Moreover, cal- cium ions by activation of calpain favor the reor- ganization of cytoskeleton, leading to detach- ment of plasmamembrane protrusions from the cortical actin [14].
Exosomes are more homogenous and smaller than shedding vesicles, with a size ranging from 30 to 120 nm and have an endosomal origin [4]. The exosomes share the biochemical char- acteristics with the internal vesicles of the mul- tivesicular bodies. It has been suggested that they are stored as intraluminal vesicles within multivesicular bodies of the late-endosome. The release of exosomes follows the fusion of mul- tivesicular bodies with the cell membrane (Figure 1B). Tetraspanins, Alix and TSG101 are considered markers of exosomes [15]. The mechanism of assembly and sorting of the exosomes is still largely unknown since a com- mon sorting signal for all cell types has not been yet identified [16]. Therefore, MVs differ on size and molecular composition depending on the
Figure 1. Production and release of shedding vesi- cles and exosomes. (A) Schematic representation of production and release from the cell surface of the shedding vesicles. Shedding vesicles are produced by budding of cell plasmamembrane. (B) Schematic representation of exosome release. The exocytic multivesicular bodies may fuse with membrane and release exosomes.
cell of origin and on the mechanism of forma- tion. In addition, the content of released MVs may vary depending on whether the secretion is constitutive or consequent to cell activation.
MVs as mediators of intercellular communica- tion
MVs released from a given cell type express on their surface the adhesion molecules of the cell of origin. Therefore, MVs may be captured through specific receptor-ligand interaction by target cells that specifically recognize them rather than just by any cell present in the micro- environment [17]. Following interaction, MVs
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Figure 2. Schematic representation of MV-mediated cell-to-cell interaction. (1) MVs may signal through surface expressed receptors leading stimulation of target cells. (2) MVs may transfer receptors from the cell of origin to the target cell. (3) MVs may transfer oncogene products, transcription factors or infectious particles to target cells. (4) MVs may mediate a hori- zontal transfer of mRNA and microRNA (miRNA) in- ducing epigenetic changes in the target cell.
may influence the behavior of the recipient cells in different ways (Figure 2).
MVs my directly stimulate the cells by a surface interaction
For instance, after activation, platelet-shed MVs coated with tissue factor (TF) are able to inter- act with molecules, such as P-selectin, ex- pressed on the surface of macrophages, poly- morphonuclear neutrophils and platelets [18]. Platelet-derived MVs remain on the surface of these cells and their phosphatydilserine en- riched membranes provide a surface for the assembly of clotting factors.
Moreover, platelet-derived MVs, may directly activate endothelial cells [19], polymorphonu- clear neutrophils [20] and monocytes [21] and influence the functions of normal and malignant human hemopoietic cells [3].
MVs may act by transferring receptors between cells
For instance, MVs can transfer the adhesion
molecule CD41 from platelets to endothelial cells [22] or to tumor cells [23] conferring them pro-adhesive properties. Other receptors that have been shown to be transferred by MVs in- clude: Fas ligand that after transferring from tumor cells to activated T cells may induce T cell apoptosis [24]; CXCR4 and CCR5 chemokine receptors that may act as co-receptors for HIV1 virus favoring the entry of the virus into cells other than the lympho-haemopoietic lineage [5, 25].
MVs may deliver proteins to target cells
For instance, it has been found that endotoxin- stimulated monocytes induce the cell death of vascular smooth muscle cells by transfer of a cell death message via encapsulated caspase-1 [26]. It has been also suggested that MVs may contribute to dissemination of certain infective agents, such as HIV or prions [27, 28]. In addi- tion tumor-derived MVs may transfer the prod- ucts of oncogenes to neighboring cells [29].
MVs may induce epigenetic changes in target cells by transferring genetic information
Tumor-derived MVs may transfer not only sur- face determinants but also mRNA of tumor cells to monocytes [30]. Indeed, MVs contain se- lected patterns of mRNA and microRNA (miRNA) associated with ribonucleoproteins involved in the intracellular traffic of RNA, suggesting a dy- namic regulation of RNA compartmentalization in MVs [31].
An epigenetic reprogramming of adult haemato- poietic stem/progenitor cells by MVs derived from murine embryonic stem cells was demon- strated by Ratajczak J et al. [32]. In these cells, MVs induce an up-regulation of early pluripotent and early hematopioetic markers and phos- phorylation of MAPKp42/44 and Akt. This bio- logic effect has been attributed to a horizontal transfer of mRNA mediated by MVs [32]. Simi- larly, Valadi et al. [33] demonstrated that exosomes from a mouse and a human mast cell line shuttle RNAs that can be transferred to other mouse and human mast cells. After trans- fer to human mast cells, new mouse proteins become detectable in the recipient cells, indi- cating that transferred exosomal mRNA can be translated after entering the target cells. We demonstrated that MVs derived from human endothelial progenitor cells (EPC) can also act
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as a vehicle for mRNA transport among cells [34]. Indeed, EPC-derived MVs activate an an- giogenic program in recipient quiescent endo- thelial cells by transferring selected patterns of mRNA. Evidence for MV-mediated transfer of genetic information has been provided by ex- periments showing translation into protein of reporter mRNA such as the green fluorescence protein mRNA (GFP) [34]. Endothelial cells tar- geted with MVs carrying GFP mRNA produce the GFP proteins [34]. In addition, we demonstrated that MVs derived from human stem cells may deliver also in vivo human mRNA to mouse cells, resulting in protein translation [35, 36]. Recently Alliotta et al. [37] demonstrated that MV entry into bone marrow cells induces tissue- specific changes in mRNA not only by direct delivery of mRNA but also by induction of tissue specific mRNA. Besides mRNA, MVs may trans- fer miRNA into target cells [31, 33, 38]. Yuan et al. [38] demonstrated that MVs derived from mouse embryonic stem cells contain abundant miRNA and that they can transfer a subset of miRNA to mouse embryonic fibroblasts in vitro. We, recently, characterized miRNA shuttled by MVs released by adult human mesenchymal stem cells [31]. Hierarchical clustering and simi- larity analysis of miRNA showed that some miRNA are selectively accumulated within MVs and absent in the cells after MV release whereas other are retained within the cells and not secreted in MVs. This suggests a regulated process of miRNA compartmentalization and secretion by MVs. Gene ontology analysis of predicted and validated targets showed that the highly expressed miRNA in MVs derived from mesenchymal stem cells may be involved in multi-organ development, cell survival, differen- tiation and immune system regulation. More- over, we demonstrated that miRNA carried by in MVs may be transferred to target cells after MV incorporation. These observations open the pos- sibility that the MV-mediated transfer of miRNA, which are naturally occurring regulators of pro- tein translation, can alter the expression of gene products in neighboring cells. Quesenberry and Aliotta [39] suggested that MV-mediated transfer of genetic information may play a criti- cal role also in stem cell biology. It has been suggested that transfer of genetic information by MVs play a pivotal role in stem cell plasticity and tissue regeneration [37, 41]. This mecha- nism possibly contributes to the paracrine ac- tion of stem cells in the repair of tissue injury [42].
Role of MVs in tumor biology
Tumor cells release large amount of MVs and the number of circulating MVs is increased in patients with cancer and correlates with poor prognosis [9]. This may depend on the pleyo- tropic effects of MVs.
Tumor induced blood coagulation and MVs
Thrombotic events are common among different cancer types and stages. In this context TF is emerging as one of the main mediators involved in the hypercoagulability of cancer patients. MVs expressing TF have a central role in trigger- ing the coagulation cascade. It has been shown that the majority of TF-bearing MVs are of tumor origin [43]. Indeed, cancer cells overexpress TF [44] and may shed it from their surface [45]. A correlation between the presence of TF-bearing MVs and a increased risk of thromboembolic events has been established [43]. The pro- coagulant properties of MVs depend also on their over-expression of phosphatydilserine pro- viding a catalytic site for the coagulation cas- cade [8, 46]. The surface expression of phos- phatydilserine facilitates the assembly and acti- vation of tenase and pro-thrombinase com- plexes, thus propagating the coagulation proc- ess [47]. Besides tumor cells, MVs in cancer patients may derive from other cells of the host, such as monocytes, erythrocytes, endothelial cells, platelets and tumor stromal cells [48]. Monocyte-derived TF-bearing MVs originate from lipid rafts and fuse with activated platelets to initiate coagulation [12]. Indeed, MVs ex- pressing P-Selectin glycoprotein ligand 1 may interact with P-Selectin-bearing platelets and increase TF-FVIIIa activation. Also MVs released from activated endothelial cells may induce thrombosis by a TF-dependent mechanism [49]. The role of MV-triggered coagulation has been implicated not only in the thromboembolic events but also in delivering signals within the tumor microenvironment, leading to prolifera- tion of dormant cancer stem cells [50, 51] or to activation of the angiogenic shift of tumors [47].
Tumor niche, angiogenesis, metastasis and MVs
Tumor growth involves a coordinated effort of different cell types to establish a favourable microenvironment. In this context, tumor cells are thought to orchestrate the behaviour of en-
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 Figure 3. Schematic representation of exchange of genetic information among tumor and stroma cells. Tumor and normal stroma cells talk each other to establish a favorable tumor niche and to promote tumor growth, invasiveness and progression via mu- tual shedding of MVs. This induces changes in the behaviour of normal cells and increases tumorigenic- ity. The MVs released from cancer cells may repro- gram the phenotype of cells present in the tumor microenvironment by delivering to cells the mRNA and/or microRNA (miRNA). This may induce epige- netic alterations in tumor endothelial cells promoting the angiogenic shift of the tumor. Moreover, tumor- derived MVs, by entering the circulation and biologi- cal fluids, may allow long distance cell-to-cell commu- nication favoring the development of a pre-metastatic niche.

dothelial cells, fibroblasts, monocytes, adipo- cytes and immune cells [47]. Besides soluble factors, MVs emerged in recent years as poten- tial candidates of cell-to-cell communication (Figure 3). An example of such role of MVs is the release by tumors of EMMPRIN a transmem- brane glycoprotein identified as a tumor-derived factor that can stimulate matrix metallopro- teinase expression in fibroblasts and conse-
quently facilitate tumor invasion and metastasis [52]. It has been shown that EMMPRIN is re- leased from the surface of tumor cells via MV shedding by a pathway dependent on protein kinase C, calcium mobilization and mitogen- activated protein kinase. These results outline a feature of tumor-stromal interaction whereby degradation of extracellular matrix by fibroblasts is controlled through the microvesicular release of EMMPRIN from tumor cells. Recently, Castel- lana et al. [47] provided evidence that tumor and normal stroma cells communicate via mu- tual shedding of MVs. This induces changes in the behaviour of normal cells and increases tumorigenicity [53]. MVs derived from prostate carcinoma cell lines express matrix metallopro- teinases (MMP) and extracellular MMP inducer at their surface, suggesting a role in extracellu- lar matrix degradation. Moreover, MVs induce the activation of fibroblasts increasing motility and resistance to apoptosis and promote MV shedding from activated fibroblasts. In turn, MVs derived from activated fibroblasts increase migration and invasion of highly metastatic prostate carcinoma cells by a mechanism, at least in part, dependent on membrane-bound CX3CL1/fractalkine ligand for chemokine recep- tor CX3CR1 [53]. Lima et al. [54] demonstrated that tumor-derived MVs modulate the establish- ment of metastatic melanomas by a mechanism dependent on phosphatidylserine, possibly by down-regulating the host’s inflammatory and immune responses.
It has been also shown that the rat pancreatic adenocarcinoma-derived MVs may form the pre- metastatic niche that allows the development of lung metastasis [55]. In this study Jung et al. [55] demonstrated that in vivo pre-metastatic changes result from cooperation of both, exosomes and soluble matrix. Recently, Al- Nedawi et al. [29] demonstrated that tumor derived MVs may act as intercellular transfer of an oncogene. Some cells of aggressive human brain tumours express a truncated and onco- genic form of the epidermal growth factor recep- tor, known as EGFRvIII. This oncogene receptor can be ‘shared’ between glioma cells by intercel- lular transfer of membrane-derived MVs, lead- ing to the transfer of oncogenic activity [29]. These experiments suggest that tumor derived MVs may allow a horizontal propagation of on- cogenes among different subsets of cancer cells thereby transforming their phenotype. The shedding of MVs from tumor cells into the sur-
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rounding environment is regulated by a small GTP-binding protein ARF6. Inhibition of ARF6 activation is associated with PKC-mediated phosphorylation of myosin light-chain, which blocks MV shedding [56]. Released MVs contain selected cellular components involved in cell adhesion and motility [56]. Wysoczynski and Ratajczak [57] demonstrated that lung cancer cells once secreted MVs in response to non- apoptotic doses of hypoxia and irradiation are able to activate and chemoattract stroma fibro- blasts and endothelial cells thus inducing the expression of several pro-angiopoietic factors in stromal cells. Moreover, stroma cells stimulated by tumor-derived MVs enhance the metastatic potential of lung cancer cells in vivo suggesting that MVs are important constituents of tumor microenvironment with a pivotal role in tumor progression, metastasis and angiogenesis. Tu- mor-derived MVs carrying hsp90a may also favor cancer cell invasion by activating plasmin [58].
Moreover, tumor-derived MVs may modulate tumor angiogenesis. MVs shed by ovarian can- cer cells may induce endothelial cell activation and angiogenesis by a CD147-mediated mecha- nism [59]. Human colorectal cancer cells ex- press TF-bearing MVs under control of activation of K-ras oncogene and inactivation of the p53 tumor suppressor, in a manner dependent on MEK/mitogen-activated protein kinase (MAPK) and phosphatidylinositol 3′-kinase (PI3K) [44]. This study established a link between the levels of MV-associated TF activity and the genetic status of cancer cells. Moreover, it suggested that TF-bearing MVs are an important effector of the K-ras-dependent tumorigenic and angio- genic phenotype in vivo. Beside TF, tumor- derived MVs may stimulate tumor growth and metastasis by promoting endothelial cell migra- tion, invasion, and tube formation, and inducing in vivo neovascularization by a mechanism de- pendent on sphingomyelin expression [60]. Tet- raspanin, a constitutive component of exosomes released by tumor cells, may also contribute to induce endothelial cell activation and angiogenesis [61]. Tetraspanin-enriched MVs have been suggested to play a central role in pre-metastatic niche preparation [62].
Al-Nedawi et al. [63] suggested that the angio- genic switch in tumors may be induced by MV- mediated transfer of the oncogenic EGFR that in turn activate an autocrine expression of VEGF.
Moreover, activated endothelial cells may com- municate at distance by transferring Delta-like 4 Notch ligand via MVs, thus propagating the an- giogenic signal [64].
Metalloproteinases harboured by endothelial MVs regulate the proteolytic activity on matrix required to elicit angiogenesis [65]. MVs shed by activated endothelial cells express on their surface matrix metalloproteinases that may allow focal proteolytic activity favoring endothe- lial cell invasion [65]. MV-associated urokinase plasminogen activator also favors the invasion of prostate cancer cells [66]. By carrying active metalloproteinases, MVs may contribute to stro- mal remodelling leading to tumor cell invasion [67]. It has been shown that CD147/ extracellular MMP inducer is expressed in MVs derived from epithelial ovarian cancer cells and that CD147-positive MVs may promote an an- giogenic phenotype in endothelial cells in vitro. The treatment of ovarian cancer cells with small interfering RNA against CD147 suppresses the angiogenic potential of MVs suggesting that vesicles shed by ovarian cancer cells may in- duce proangiogenic activities by a CD147- mediated mechanism [59].
Data relative to the role of EPC in tumor angio- genesis remain controversial [68]. Some experi- mental studies have shown that EPC are capa- ble of incorporating and differentiating in vessel -like structures contributing to tumor angiogene- sis [69, 70]. However, other studies suggested that EPC promote angiogenesis without a direct contribution in the formation of vessels with only a perivascular localization without incorpo- ration into the vessel wall [71, 72]. In the hu- man tumor vasculature, co-expression of Y chro- mosome and endothelial markers varies from 1 up to 12%, depending on the tumor type in pa- tients submitted to bone marrow transplanta- tion from donors of opposite sex [73]. There- fore, it has been suggested that EPC stimulate in a paracrine manner resident endothelial cells [68, 73]. In line with this possibility, we found that MVs released by human EPC activate an angiogenic program in normal endothelial cells by a horizontal transfer of mRNA [34]. In vitro, after incorporation in normal endothelial cells by interaction with α4- and β1-integrins ex- pressed on MV surface, MVs promote endothe- lial cell survival, proliferation and organization in capillary-like structures [34]. This effect also occurs in vivo in SCID mice, where MV-
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stimulated human endothelial cells implanted subcutaneously within Matrigel organize in a patent vessel network connected with the mur- ine vasculature. Pre-treatment of MVs with ele- vated concentration of RNase reduces their angiogenic activity suggesting a critical role for RNA transfer following MV incorporation. The angiogenic effect correlates with transfer of mRNA following the MV incorporation within the normal endothelial cells after adhesion receptor -mediated interaction. The molecular analysis of mRNA indicates that MVs derived from EPC shuttle a specific subset of cellular mRNA, in- cluding mRNA associated with pathways rele- vant to angiogenesis such as the PI3K/AKT and eNOS signaling pathways. Protein expression and functional studies demonstrated that PI3K and eNOS are up-regulated in target cells after MV incorporation. The transfer of genetic infor- mation by MVs has been also shown for MVs derived from tumor cells (Figure 3). Indeed, MVs released by colorectal cancer cells are enriched in cell cycle-related mRNAs and promote endo- thelial cell proliferation, suggesting that tumor derived MVs may be involved in the angiogenic shift and in tumor progression [74]. Skog et al. [75] demonstrated that MVs released by glioblastoma tumour contain selected patterns of mRNA, miRNA and angiogenic proteins. These MVs after incorporation by normal host cells, such as brain microvascular endothelial cells, transfer mRNA for a reporter protein that can be translated by the recipient cells. More- over, these MVs carry angiogenic proteins and in vitro induce tube formation by endothelial cells. This study confirms that MVs released by cancer cells may deliver specific and functional RNAs and proteins to recipient cells in the tu- mor microenvironment. In fact, recent studies suggest epigenetic mutations in stroma and tumor-derived endothelial cells [76].
Tumor-derived MVs in immuno-escape and chemoresistance
Recently, the role of MVs in immune response has been extensively reviewed by Thery C et al. [77]. Depending on the cell of origin and the molecular composition MVs may either stimu- late or inhibit the immune response. Indeed, MVs may act as direct peptide-MHC complex presentation to T cell, may transfer antigen or peptide-MHC complex to dendritic cells leading to indirect antigen presentation, may activate natural killer cells and macrophages or confer
protection of T cells against “activation-induced cell death”. Moreover, CD40L bearing MVs may stimulate B cell activation and antibody produc- tion [78]. Based on these properties MVs de- rived from mature dendritic cells have been used as vaccines to stimulate efficient antitu- mor cytotoxic T-lymphocyte response [79, 80].
On the other hand MVs may inhibit immune re- sponse favoring escape of tumor cells from im- mune surveillance. This may depend on the ability of tumor-derived MVs to induce apoptosis in activated anti-tumor T cells, impairment of monocyte differentiation into dendritic cells and induction of myeloid suppressive cells [10, 81]. Moreover, vesicular shedding of terminal com- ponents of complement from the cell plasma membrane [82] by a mechanism called “complement resistance” may confer protection to tumor cells from antibody mediated immune response. Similarly shedding of Fas ligand from tumor cell surface reduces sensitivity to T cell Fas-mediated apoptosis [83].
MVs may also facilitate tumor cell survival by expulsion of therapeutic drugs from cancer cells. A correlation between MV release and multidrug resistance has been established. Can- cer cells resistant to chemotherapy were found to release significant more MVs than those sen- sitive to chemotherapy [84]. In addition, these MVs contained significant more chemotherapic drugs than those derived from cancer cells sen- sitive to chemotherapy. Based on this observa- tion it has been suggested that chemotherapeu- tic agents may be extruded from cells via MVs [85].
MVs as diagnostic tool in cancer
The proteomic and the cytofluorimetric analysis [86, 87] of MVs present in the circulation may provide qualitative and quantitative information on MVs present in blood. It has been suggested that mucin-bearing MVs may be useful in the early detection of adenocarcinomas [88, 89]. In glioblastomas the tumor-specific EGFRvIII marker is expressed by MVs and may provide diagnostic information [75]. The proteomic analysis on urinary MVs allowed identification of potential biomarkers of bladder cancer [90, 91]. Using a sandwich ELISA to capture and quantify exosomes in plasma Logozzi et al. [92] demon- strated high levels of exosomes expressing CD63 and caveolin-1 in plasma of patients with
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melanoma. Several studies have suggested that the level of circulating MVs correlate with the prognosis and survival of patients with cancer [9, 93].
The recent identification that MVs carry specific patterns of mRNA and miRNA stimulate re- search aimed to find a molecular signature of circulating MVs in cancer patients that may be relevant for diagnostic or prognostic purpose. Cancer-specific mRNA has been detected in circulating MVs in patients with glioblastoma [75], in gastric cancer [94] and in breast cancer patients [95]. The profile of miRNA carried by MVs may also be exploited for diagnostic pur- poses [96]. miRNA in plasma would be de- graded by RNase if not protected by a mem- brane envelope. This envelope is provided by MVs that therefore may allow detection of miRNA in the circulation. The secretory mecha- nisms and intercellular transfer of miRNA via MVs have been recently studied by Kosaka N et al. [97]. Hunter et al. [98] defined the miRNA expression in circulating plasma MVs in normal subjects. Cancer-specific miRNA have been de- tected in patients with ovarian cancer showing that miRNA profile may vary with the disease stage [99]. Studies on lung cancer demon- strated that the profile of circulating miRNA pre- sent in MVs is similar to that of tumor-derived miRNA, suggesting that miRNA detectable in MVs might be useful as a screening test for lung adenocarcinoma [100]. miR-21 which is over- expressed in glioblastoma is also detectable in the circulating MVs [75]. Overall these studies suggest that gaining more information on the molecular composition of MVs may offer new diagnostic and prognostic information. More- over, Renzulli et al. [101], on the base of the observation that prostate cancer tumor cells may induce via MVs prostate specific gene ex- pression in circulating monocytes, stem cells or other cells, altering their phenotype, proposed novel therapeutic strategies to block the MV release from cancer cells or the MV entry in re- cipient cells.
Conclusion
In conclusion, MVs have pleiotropic biological actions implicated in signalling among cells and in transferring gene products. The biological action of MVs may take place either in a defined microenvironment either in long distance as they may enter the circulation and other biologi-
cal fluids. The presence of MVs in body fluids, their number, cellular origin, composition and function, can depend on disease state and represents a new non-invading potential diag- nostic tool. The finding that MVs may exchange genetic information among cells suggests that tumor and stromal cells talk each other to es- tablish a favorable tumor niche and to promote tumor growth, invasiveness and progression.
Moreover, the fact that MVs contain genetic material under the form of RNA, may allow an easy screening for cancer genetic markers. The identification of signals delivered by MVs may also open new therapeutic strategies. In particu- lar, the removal from plasma of harmful MVs may be beneficial in tumors where MVs deliver thrombogenic and cell transforming signals.
Disclosure
Stefania Bruno is a full time employee of Sis- Ter, Palazzo Pignano; Ciro Tetta is a full time employee of Fresenius Medical Care. All the other authors declared no competing interests.
Acknowledgements
Our research is supported by grants from Re- gione Piemonte, Piattaforme Biotecnologiche, project PiSTEM and Oncoprot and from Minis- tero dell’Istruzione, dell’Università e della Ricrca (MIUR) project PRIN08.
Please address correspondence to: Giovanni Camussi, PhD, Dipartimento di Medicina Interna, Ospedale Maggiore S. Giovanni Battista, Corso Dogliotti 14, 10126, Torino, Italy; Phone +39-011-
Microvesicles and cancer
6336708, Fax +39-011-6631184, giovanni.camussi@unito.it
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