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Published OnlineFirst March 31, 2016; DOI: 10.1158/0008-5472.CAN-15-2804
Cancer
Research
Review
Implications of Extracellular Vesicle Transfer on
Cellular Heterogeneity in Cancer: What Are the
Potential Clinical Ramifications?
Anoek Zomer and Jacco van Rheenen
Abstract
The functional and phenotypic heterogeneity of tumor cells
represents one of the greatest challenges in the successful treatment of cancer patients, because it increases the risk that certain
individual tumor cells possess the ability to, for example, metastasize or to tolerate cytotoxic drugs. This heterogeneity in cellular
behavior is driven by genetic and epigenetic changes and environmental differences. Recent studies suggest that an additional
layer of complexity of tumor heterogeneity exists, based on
the ability of cells to share functional biomolecules through local
and systemic transfer of extracellular vesicles (EV), with profound
effects on cellular behavior. The transfer of functional biomolecules between various populations of tumor cells and between
tumor cells and nontumor cells has large consequences for both
the tumor cells and the microenvironment that support the
cellular behavior of tumor cells, and therefore for the clinical
outcome of cancer. Here, we discuss the latest findings on EV
transfer and the potential implications of EV-mediated local and
systemic transmission of phenotypic behavior, particularly in the
context of tumor heterogeneity, metastatic disease, and treatment
response. Cancer Res; 76(8); 2071–5. 2016 AACR.
Introduction
over time, resulting in continuously changing nongenetic tumor
heterogeneity. Soluble factors such as secreted ligands that can
bind plasma membrane receptors of both tumor cells and stromal
cells have been extensively studied and are considered to be the
central players in the communication between different cell types.
For example, it has been shown that (epi)genetically distinct
tumor cell subsets enhance tumor growth (6, 7) or invasion
(8) by transferring soluble proteins from one cell population to
another. In recent years, it has become increasingly clear that cells
also use other mechanisms to facilitate intercellular exchange of
cellular signals including cell–cell fusion (9), gap junctions (10),
tunneling nanotubes (11), and extracellular vesicles (12). In this
review, we focus on the recent findings that suggest that EVs
influence tumor heterogeneity, thereby regulating multiple
aspects of tumor progression, and we discuss the potential clinical
implications.
Most cancers consist of a complex mixture of tumor cells,
nontransformed stromal cells and noncellular factors including
growth factors and the extracellular matrix (1). Tumor cells
themselves also display a large genetic heterogeneity due to
genomic instability that leads to an increased mutation rate and
the emergence of different somatic aberrations (2). The behavior
of individual tumor cells is not solely determined by genetic
alterations. Heterogeneity between tumor cells can be further
increased due to regional differences in the tumor microenvironment (TME), for example, in hypoxia or the presence of growth
factors, which drives heterogeneous expression profiles even in
genetically homogenous cells (1, 3, 4). The genetic and nongenetic tumor heterogeneity creates a large variation in behavior of
tumor cells. The existence of these different cell populations
currently represents one of the greatest challenges in successfully
treating cancer patients since it increases the chance that certain
individual tumor cells possess the ability to, for example, metastasize or tolerate cytotoxic drugs. Intravital microscopy studies
showed that the presence of only a few individual cells with these
types of traits within a tumor is sufficient to drive processes such as
cancer progression and metastasis (5).
Because the TME is highly dynamic and constantly modulated
during tumor progression, the interactions between different cell
types and noncellular microenvironmental factors also change
Cancer Genomics Netherlands, Hubrecht Institute-KNAW and University Medical Center Utrecht, Utrecht, the Netherlands.
Corresponding Author: Jacco van Rheenen, Hubrecht Institute, Uppsalalaan 8,
Utrecht, 3584CT, the Netherlands. Phone: 31-30-2121905; Fax: 31-30-251-64-64;
E-mail: [email protected]
doi: 10.1158/0008-5472.CAN-15-2804
2016 American Association for Cancer Research.
Cell–Cell Communication through the
Transfer of Extracellular Vesicles
Almost all cells release different types of extracellular vesicles.
These are typically classified as either microvesicles, which bud
from the cellular plasma membrane, or exosomes, which originate
from multivesicular bodies and shed upon fusion of multivesicular bodies with the plasma membrane (12). Since it is not clear
yet whether the various vesicles that are present in an in vivo
microenvironment have distinct or overlapping functions, we use
the generic term extracellular vesicles (EV) in this review. In cancer
patients, tumor-derived EVs have been found in diverse body
fluids such as blood (13) and ascites (14). It is increasingly evident
that EVs have a key role in regulating cell–cell communication
through the transfer of molecular information (12). An early
landmark paper showed that EVs released by mast cells contain
messenger (m)RNA molecules, and that these transcripts can
be transported to recipient cells where they are translated into
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2071
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Zomer and van Rheenen
functional proteins (15). Besides mRNA, EVs are now known to
shuttle many other functional biomolecules, including proteins,
lipids, microRNA (miRNA), and DNA, between different cell types
present in multicellular organisms (12). Transfer of vesicular
content can influence the phenotypic behavior or fate of recipient
cells, for example, by inducing differentiation or de-differentiation, or by promoting apoptosis (reviewed in ref.16). Although it
is currently not well understood how long EV-mediated effects last,
the duration may be dependent on the type of biomolecules being
transferred. Possibly, vesicular transfer of DNA, miRNA, mRNA,
and/or transcription factors leads to epigenetic reprogramming of
recipient cells, resulting in a stable behavioral change. Conversely,
EV-mediated transfer of, for example, membrane receptors, proteins that are considered to actively turnover, may only alter the
phenotype of recipient cells for a short period of time.
Studying the Uptake of EVs Released by
Experimentally Defined Cells
Accumulating in vitro evidence suggests that EVs contribute to
the heterogeneous nature of tumors by transferring molecular
information between different cell types or cells with differential
behavior. However, it has been challenging to fully characterize
these individual cells in an in vivo setting, because in vivo only a few
cells may take up EVs and no techniques existed to distinguish
those cells from the vast majority of cells that do not take up EVs.
Moreover, since virtually all cells produce EVs, it has been challenging to study the consequence of the uptake of EVs that are
produced by specific cell types. Recently, the Cre-LoxP system was
utilized to overcome these technical limitations and to directly
identify tumor cells that are exposed to the content of EVs released
by a researcher-defined population of cells in living mice (17–19).
In this Cre-LoxP system, cells express a reporter construct that
switches from DsRed to eGFP expression upon the functional
transfer of Cre-recombinase (Cre) from Cre-expressing cells
(19). Cells that have and have not taken up EVs from Cre-expressing cells can be discriminated on the basis of their color (green
versus red) and therefore this system allows the identification of
reporter cells that are exposed to the cargo of only the EVs released
by Cre-expressing cells. Differential behavior of these two cell
populations (that have or have not taken up EVs) can be studied
to unravel functional implications of EV transfer. Importantly, and
in contrast with other reporter systems that make use of fusing
fluorescent proteins to EV and/or membrane markers (20, 21), the
Cre-LoxP system reports both the uptake and the functional release
of the cargo of EVs. Furthermore, the Cre-LoxP method provides
the ability to detect several modes of Cre exchange between Creexpressing cells and reporter-expressing cells including cell–cell
fusion and the transfer of biomolecules through direct cell–cell
contact, although to our knowledge it has, thus far, only been used
to study EV exchange (17–19). We have recently used this method
to show that more malignant cells phenocopy their behavior to less
malignant cells through EV transfer in living mice (19).
Phenocopying of Malignant Behavior
through the Transfer of EVs
High-resolution intravital microscopy has been extensively
used to study the heterogeneous behavior of cancer cells within
tumors (5). We used intravital microscopy to study the release of
EVs from tumor cells in vivo and showed that highly migratory
2072 Cancer Res; 76(8) April 15, 2016
MDA-MB-231 breast tumor cells release a heterogeneous population of small and large EVs into the TME (19). Next, we isolated
these EVs and tumor cells and identified that the mRNAs of
around 200 genes involved in metastasis are enriched in these
EVs. Because it has already been shown that heterogeneous
tumors consist of subpopulations of cells with differing metastatic
potential (22), we questioned whether the "malignant" message
encoded in EVs released by highly migratory tumor cells could be
transmitted to other, less aggressive tumor cells. To address this,
we used the Cre-LoxP system and tested whether the less malignant T47D reporter cells report the uptake of EVs released by Creexpressing highly malignant MDA-MB-231 cells (19). Interestingly, we observed that EVs released by MDA-MB-231 cells are
taken up by the T47D cells located within the same tumor and
within distant tumors, suggesting local and systemic EV transfer.
Next, we combined intravital imaging to visualize cellular behavior of individual cells and the Cre-LoxP system to distinguish
between cells that did or did not take up EVs released by Creexpressing cells. By intravital imaging, we showed that T47D cells
that report the uptake EVs released by the aggressive Cre-expressing MDA-MB-231 cells display an increased migratory behavior
compared with T47D cells that do not report the uptake of these
EVs. Since migration is one of the limiting steps in the metastatic
cascade, we next tested whether metastasis was also affected. In
line with the enhanced migration, we found that the capacity to
form lung metastases increases for those T47D cells that report the
uptake of EVs released by the highly metastatic cells compared
with their counterparts that did not take up EVs. These findings
suggest that within primary tumors, cells with a high metastatic
potential transfer EVs to less metastatic cells, thereby equipping
them with migratory properties and allowing these cells to leave
the primary tumor.
EV Transfer May Contribute to Tumor
Progression by Directing the Behavior of
Multiple Cell Types
The existence of in vivo EV transfer between tumor cells and
between tumor and nontransformed cells (18, 19) adds another
layer of complexity to the concept of cell–cell communication and
tumor heterogeneity. The functional effects of EV transfer between
different cell types present in tumors have been extensively
investigated in vitro. First, EVs may mediate cross-talk between
different subsets of cancer cells. A key study showed that a highly
aggressive population of glioma cells, expressing a truncated and
oncogenic form of mutated EGFR (EGFRVIII), can transfer this
protein to EGFRVIII-negative glioma cells via EVs (23). This
transfer leads in the recipient cells to an increased oncogenic
activity including activation of the mitogenic MAPK and survival
Akt pathway, and an increase in anchorage-independent growth.
Similarly, Higginbotham and colleagues showed that tumor EVs
containing EGFR ligands can increase the invasiveness of recipient
cancer cells (24). Second, EVs released by tumor cells have been
shown to be capable of transforming healthy nontumorigenic
cells. For example, breast cancer EVs containing RISC-associated
miRNAs mediate silencing of target mRNAs in nontumorigenic
recipient cells, thereby changing the transcriptome of these cells
and instigating malignant transformation (25). Other studies
have shown that tumor EVs can be transferred to nontransformed
stromal cells such as endothelial cells or immune cells to shape the
TME and accelerate tumor progression. In vitro evidence suggests
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Copying Metastatic Behavior through Extracellular Vesicles
Primary tumor
Metastasis
Extracellular
vesicle exchange
Figure 1.
Reciprocal EV-mediated
communication between cells in
primary tumors and distant
metastases. Different cell types
present in heterogeneous tumors,
including tumor cells and stromal
cells, release EVs that can be taken up
by neighboring cells or can be
systemically transferred to distant
cells present in the (pre)metastatic
niche. Conversely, cells present in
metastases may release EVs that
change the behavior of cells in the
primary tumor. In this way, removal of
the primary tumor or metastases,
for example, by surgery or
chemotherapy, may affect the
behavior of cells at distant sites in an
EV-dependent manner.
Lung
Tumor cells
Stromal cells
Extracellular vesicles
Endothelial cells
Extracellular matrix
© 2016 American Association for Cancer Research
that EVs released by tumor cells have the potential to induce
angiogenesis (26, 27) and inhibit antitumor immune responses
(reviewed in ref.28). These results have been recently validated in
in vivo experiments using the Cre-LoxP system, showing that
tumor EVs are taken up by myeloid-derived suppressor cells
(MDSC) present in the local TME (18). MDSCs that take up these
EVs display enhanced immunosuppressive properties compared
with MDSCs that do not take up tumor EVs. Third, several studies
demonstrated functional EV transfer from stromal cells to tumor
cells. For example, EVs released by fibroblasts and activated T cells
have been shown to increase the invasive phenotype of tumor
cells (29, 30).
Collectively, these studies suggest that EV transfer between
different cell types (transformed and nontransformed) can
influence tumor progression. However, it is important to validate these findings under physiologic conditions where both
EV release and EV uptake occur in an in vivo setting. Recent
reports, showing EV exchange between tumor cells and CD45þ
leukocytes or CD45 cells in living mice by means of the CreLoxP system (18, 19), illustrate the great potential of this
approach to further characterize the molecular and functional
effects of in vivo EV transfer between the many different cell
types that are present in heterogeneous tumors and in distant
organs (Fig. 1).
Systemic EV Transfer
EVs released in vivo by primary tumor cells have been shown to
travel through body fluids, in this way conveying functional
information to physically separated primary tumor cells and to
distant nontransformed stromal cells (Fig. 1). A few pivotal
studies showed that systemically administrated tumor EVs can
modulate distant stromal cells to facilitate metastatic outgrowth
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in distant organs (31–36). First, EVs released from highly metastatic melanoma cells were shown to prepare a premetastatic
niche in the lungs and bone (31). Upon injection in naive mice,
these EVs are taken up by bone marrow-derived cells, resulting in
the "education" of these cells by increasing their amounts of
MET and activated phospho-MET. The reprogrammed bone marrow-derived cells display a provasculogenic phenotype and mobilize to distant organs to prepare them for the arrival of tumor cells
by promoting angiogenesis formation. A second study by Lyden
and colleagues showed that pancreatic cancer-derived EVs can
prime the liver for metastatic spread (32). EVs derived from a
pancreatic cancer cell line are taken up by liver-resident Kupffer
cells with subsequent effects on their gene expression profile. In
response to an increased TGFb release by Kupffer cells, hepatic
stellate cells increase the production of fibronectin, enhancing
the recruitment of macrophages from the bone marrow. These
pancreatic cancer EVs-induced changes in the liver microenvironment facilitate the outgrowth of tumor cells that escape
from the primary tumor and increased liver metastatic burden.
These results are supported by a study showing that cancerderived EVs can be taken up by distant macrophages that
subsequently activate NF-kB and secrete proinflammatory cytokines (33). Another recent paper showed that tumor-derived
EVs can suppress glucose uptake by nontumor cells in the
premetastatic niche thereby increasing nutrient availability for
metastasized tumor cells (34). In addition to these studies
showing that tumor-derived EVs can create a premetastatic
niche by instructing stromal cells to create a favorable environment for arriving tumor cells, other evidence indicates
that EVs released by tumor cells can promote destruction of
the vasculature in distant organs, including the blood–brain
barrier, thereby facilitating the migration of tumor cells
through these natural barriers (35, 36).
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Zomer and van Rheenen
Taken together, these studies have revealed that tumor-derived
EVs can modulate distant stromal cells to favor the outgrowth of
metastases. Recent experiments utilizing the Cre-LoxP method
confirmed these results by showing that nontransformed cells in
the lungs, spleen and lymph nodes take up EVs directly released
from tumor cells in vivo (19). Future studies should determine
whether the potential of EVs to travel far from their site of origin is
only relevant for creating a premetastatic niche or whether this
feature also contributes to later stages of metastasis formation.
Potential Clinical Implications of Systemic
EV Transfer
A vast amount of literature suggests that complex and bidirectional interactions occur between the primary tumor and metastatic
foci (37). For example, a number of clinical case studies suggest that
surgical removal of the primary tumor can accelerate metastatic
development (e.g., refs. 38, 39). Although serum transplantation
experiments between animals provided evidence for a serum
growth factor being responsible for communication between primary tumor and metastatic cells (40), the exact molecular mechanism underlying the clinical observations is still unknown. Potentially, part of this cross-talk could occur through systemic transfer of
EVs (Fig. 1). Tumor cells that are able to metastasize to a secondary
site may transfer EVs and increase the metastatic capacity of other
tumor cells in the primary tumor, thereby creating a feed-forward
loop reinforcing metastasis formation. In line with this idea, two
recent studies showed that systemic delivery of EVs derived from
highly metastatic tumors can enhance metastasis formation of
otherwise weakly metastatic cells (19, 41). Conversely, EVs released
by less-malignant primary tumor cells may slow down metastatic
outgrowth, thereby explaining clinical observations that report an
acceleration of metastatic outgrowth upon removal of the primary
tumor (37). Intravital imaging experiments demonstrated that
cancer cells that have taken up EVs released by weakly metastatic
tumor cells display a small decrease in in vivo migration compared
with cells that did not take up these EVs, indicating that less
malignant behavior also has the potential to be phenocopied
through the transfer of EVs (19).
Anticancer Therapies Are Potentially
Influenced by EV Transfer
A relatively new area of investigation involves the relationship
between EV transfer and response to anticancer treatments, and so
far in only a few studies, this clinically relevant subject was
approached. EVs can mediate drug resistance in a direct manner
through uptake of drugs in vesicles thereby limiting the bioavailability of the drug to tumor cells. Especially under low pH
conditions, tumor cells release cisplatin-containing EVs, resulting
in lower amounts of cisplatin in tumor cells (42). Apart from the
sequestration of drugs in vesicles, EVs can be involved in mediating drug resistance in an indirect manner by transferring biomolecules to recipient cells that direct these cells toward a drug
resistant phenotype. In vitro experiments showed that chemoresistant cells can confer resistance on chemosensitive cells through
EVs by, for example, stimulating multidrug efflux transporter
P-glycoprotein production (43, 44) or by stimulating the prosurvival AKT/mTOR pathway (45). Stromal cells present in the TME
have also shown to promote tumor cell survival after radiation or
chemotherapy through the transfer of EVs (46).
2074 Cancer Res; 76(8) April 15, 2016
It is not extensively explored whether functional biomolecules
can be transferred through apoptotic bodies, a type of EVs formed
upon programmed cell death, thereby mediating cross-talk
between the many different cell types in tumors especially upon
therapy. Apoptotic bodies were previously considered as "waste
bags" containing the expelled content of dying cells, but findings
from the last decade challenged this notion as it became clear that
also apoptotic bodies are engulfed by other cells with functional
effects on behavior. For example, two studies showed that apoptotic bodies released by transformed cells can horizontally
transfer DNA to fibroblasts and endothelial cells and that the
DNA can be propagated upon division of recipient cells, provided
that the cells are p53 negative (47, 48). As in 50% of all human
tumors, the p53 gene is mutated or deleted, DNA transfer through
apoptotic bodies may be extremely relevant in a tumor setting,
and may have profound consequences for tumor progression.
Especially upon treatment, dying tumor cells may spread DNA
and mutations to surviving tumor cells, thereby accelerating the
process of tumor evolution. Also, it has been shown that concomitant transfer of tumor DNA with genes that inactivate the p53
pathway allows tumor DNA propagation in nontransformed cells
(48), indicating that apoptotic tumor cells could affect the behavior of nontransformed cells. However, in-depth in vivo studies are
essential to test such hypotheses in a physiologic setting and to
fully understand the clinical implications of the transfer of apoptotic bodies.
Concluding Remarks
A plethora of studies that used in vitro-isolated EVs emphasize
the importance of EV-mediated cross-talk between cells in tumor
progression and response to anticancer treatments. These data in
combination with the recent in vivo evidence of EV transfer (18, 19,
31, 32) revolutionized our ideas on cell–cell communication in
heterogeneous tumors. Future studies, for example, using the CreLoxP system, should focus on validating in vitro findings under
physiologic conditions where both EV release and EV uptake take
place in an in vivo setting. Moreover, future studies that will reveal
the molecular mechanisms of EV loading, release and uptake of
the various subtypes such as microvesicles and exosomes, will
further help to investigate the role and relevance of EV transfer in
cancers. Together, these findings will significantly contribute to
understanding the complex communication mechanisms that
take place in heterogeneous tumors and will determine the
clinical implications of EV transfer.
Disclosure of Potential Conflicts of Interest
No potential conflicts of interest were disclosed.
Acknowledgments
The authors thank all members of the van Rheenen group for critically
reading this manuscript.
Grant Support
This work was supported by a VIDI fellowship (91710330); research grants
from the Dutch Organization of Scientific Research (NWO; 823.02.017), the
Dutch Cancer Society (KWF; HUBR 2009-4621) and Cancer Genomics Netherlands (all J. van Rheenen); and equipment grants (175.010.2007.00 and
834.11.002) from the Dutch Organization of Scientific Research (NWO).
Received October 8, 2015; revised December 11, 2015; accepted December
11, 2015; published OnlineFirst March 31, 2016.
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Copying Metastatic Behavior through Extracellular Vesicles
References
1. Junttila MR, de Sauvage FJ. Influence of tumour micro-environment
heterogeneity on therapeutic response. Nature 2013;501:346–54.
2. Swanton C. Intratumor heterogeneity: evolution through space and time.
Cancer Res 2012;72:4875–82.
3. Shahrzad S, Bertrand K, Minhas K, Coomber BL. Induction of DNA
hypomethylation by tumor hypoxia. Epigenetics 2007;2:119–25.
4. Hamm CA, Stevens JW, Xie H, Vanin EF, Morcuende JA, Abdulkawy H, et al.
Microenvironment alters epigenetic and gene expression profiles in Swarm
rat chondrosarcoma tumors. BMC Cancer 2010;10:471.
5. Ellenbroek SI, van Rheenen J. Imaging hallmarks of cancer in living mice.
Nat Rev Cancer 2014;14:406–18.
6. Cleary AS, Leonard TL, Gestl SA, Gunther EJ. Tumour cell heterogeneity
maintained by cooperating subclones in Wnt-driven mammary cancers.
Nature 2014;508:113–7.
7. Inda MM, Bonavia R, Mukasa A, Narita Y, Sah DW, Vandenberg S, et al.
Tumor heterogeneity is an active process maintained by a mutant EGFRinduced cytokine circuit in glioblastoma. Genes Dev 2010;24:1731–45.
8. Calbo J, van Montfort E, Proost N, van Drunen E, Beverloo HB, Meuwissen
R, et al. A functional role for tumor cell heterogeneity in a mouse model of
small cell lung cancer. Cancer Cell 2011;19:244–56.
9. Ogle BM, Cascalho M, Platt JL. Biological implications of cell fusion. Nat
Rev Mol Cell Biol 2005;6:567–75.
10. Alexander DB, Goldberg GS. Transfer of biologically important molecules
between cells through gap junction channels. Curr Med Chem 2003;10:
2045–58.
11. Rustom A, Saffrich R, Markovic I, Walther P, Gerdes HH. Nanotubular
highways for intercellular organelle transport. Science 2004;303:1007–10.
12. Raposo G, Stoorvogel W. Extracellular vesicles: exosomes, microvesicles,
and friends. J Cell Biol 2013;200:373–83.
13. Melo SA, Luecke LB, Kahlert C, Fernandez AF, Gammon ST, Kaye J, et al.
Glypican-1 identifies cancer exosomes and detects early pancreatic cancer.
Nature 2015;523:177–82.
14. Andre F, Schartz NE, Movassagh M, Flament C, Pautier P, Morice P, et al.
Malignant effusions and immunogenic tumour-derived exosomes. Lancet
2002;360:295–305.
15. Valadi H, Ekstrom K, Bossios A, Sjostrand M, Lee JJ, Lotvall JO. Exosomemediated transfer of mRNAs and microRNAs is a novel mechanism of
genetic exchange between cells. Nat Cell Biol 2007;9:654–9.
16. Mittelbrunn M, Sanchez-Madrid F. Intercellular communication: diverse
structures for exchange of genetic information. Nat Rev Mol Cell Biol 2012;
13:328–35.
17. Ridder K, Keller S, Dams M, Rupp AK, Schlaudraff J, Del Turco D, et al.
Extracellular vesicle-mediated transfer of genetic information between the
hematopoietic system and the brain in response to inflammation. PLoS
Biol 2014;12:e1001874.
18. Ridder K, Sevko A, Heide J, Dams M, Rupp AK, Macas J, et al. Extracellular
vesicle-mediated transfer of functional RNA in the tumor microenvironment. Oncoimmunology 2015;4:e1008371.
19. Zomer A, Maynard C, Verweij FJ, Kamermans A, Schafer R, Beerling E, et al.
In Vivo imaging reveals extracellular vesicle-mediated phenocopying of
metastatic behavior. Cell 2015;161:1046–57.
20. Lai CP, Kim EY, Badr CE, Weissleder R, Mempel TR, Tannous BA, et al.
Visualization and tracking of tumour extracellular vesicle delivery and RNA
translation using multiplexed reporters. Nat Commun 2015;6:7029.
21. Sung BH, Ketova T, Hoshino D, Zijlstra A, Weaver AM. Directional cell
movement through tissues is controlled by exosome secretion. Nat Commun 2015;6:7164.
22. Fidler IJ. Tumor heterogeneity and the biology of cancer invasion and
metastasis. Cancer Res 1978;38:2651–60.
23. Al-Nedawi K, Meehan B, Micallef J, Lhotak V, May L, Guha A, et al.
Intercellular transfer of the oncogenic receptor EGFRvIII by microvesicles
derived from tumour cells. Nat Cell Biol 2008;10:619–24.
24. Higginbotham JN, Demory Beckler M, Gephart JD, Franklin JL, Bogatcheva
G, Kremers GJ, et al. Amphiregulin exosomes increase cancer cell invasion.
Curr Biol 2011;21:779–86.
25. Melo SA, Sugimoto H, O'Connell JT, Kato N, Villanueva A, Vidal A, et al.
Cancer exosomes perform cell-independent microRNA biogenesis and
promote tumorigenesis. Cancer Cell 2014;26:707–21.
www.aacrjournals.org
26. Skog J, Wurdinger T, van Rijn S, Meijer DH, Gainche L, Sena-Esteves M, et al.
Glioblastoma microvesicles transport RNA and proteins that promote tumour
growth and provide diagnostic biomarkers. Nat Cell Biol 2008;10:1470–6.
27. Al-Nedawi K, Meehan B, Kerbel RS, Allison AC, Rak J. Endothelial expression of autocrine VEGF upon the uptake of tumor-derived microvesicles
containing oncogenic EGFR. Proc Natl Acad Sci U S A 2009;106:3794–9.
28. Robbins PD, Morelli AE. Regulation of immune responses by extracellular
vesicles. Nat Rev Immunol 2014;14:195–208.
29. Cai Z, Yang F, Yu L, Yu Z, Jiang L, Wang Q, et al. Activated T cell exosomes
promote tumor invasion via Fas signaling pathway. J Immunol 2012;188:
5954–61.
30. Luga V, Zhang L, Viloria-Petit AM, Ogunjimi AA, Inanlou MR, Chiu E, et al.
Exosomes mediate stromal mobilization of autocrine Wnt-PCP signaling
in breast cancer cell migration. Cell 2012;151:1542–56.
31. Peinado H, Aleckovic M, Lavotshkin S, Matei I, Costa-Silva B, Moreno-Bueno
G, et al. Melanoma exosomes educate bone marrow progenitor cells toward a
pro-metastatic phenotype through MET. Nat Med 2012;18:883–91.
32. Costa-Silva B, Aiello NM, Ocean AJ, Singh S, Zhang H, Thakur BK, et al.
Pancreatic cancer exosomes initiate pre-metastatic niche formation in the
liver. Nat Cell Biol 2015;17:816–26.
33. Chow A, Zhou W, Liu L, Fong MY, Champer J, Van Haute D, et al. Macrophage
immunomodulation by breast cancer-derived exosomes requires Toll-like
receptor 2-mediated activation of NF-kappaB. Sci Rep 2014;4:5750.
34. Fong MY, Zhou W, Liu L, Alontaga AY, Chandra M, Ashby J, et al. Breastcancer-secreted miR-122 reprograms glucose metabolism in premetastatic
niche to promote metastasis. Nat Cell Biol 2015;17:183–94.
35. Zhou W, Fong MY, Min Y, Somlo G, Liu L, Palomares MR, et al. Cancersecreted miR-105 destroys vascular endothelial barriers to promote metastasis. Cancer Cell 2014;25:501–15.
36. Tominaga N, Kosaka N, Ono M, Katsuda T, Yoshioka Y, Tamura K, et al.
Brain metastatic cancer cells release microRNA-181c-containing extracellular vesicles capable of destructing blood-brain barrier. Nat Commun
2015;6:6716.
37. Demicheli R, Retsky MW, Hrushesky WJ, Baum M, Gukas ID. The effects of
surgery on tumor growth: a century of investigations. Ann Oncol 2008;19:
1821–8.
38. Sugarbaker EV, Thornthwaite J, Ketcham AS. Inhibitory effect of a primary
tumor on metastasis. In: Day SB, Myers WPL, Stansly P, editors. Progress in
cancer research and therapy . New York, NY: Raven Press; 1977. 227–40.
39. Folkman J. Angiogenesis in cancer, vascular, rheumatoid and other disease.
Nat Med 1995;1:27–31.
40. Fisher B, Gunduz N, Coyle J, Rudock C, Saffer E. Presence of a growthstimulating factor in serum following primary tumor removal in mice.
Cancer Res 1989;49:1996–2001.
41. Le MT, Hamar P, Guo C, Basar E, Perdigao-Henriques R, Balaj L, et al. miR200-containing extracellular vesicles promote breast cancer cell metastasis.
J Clin Invest 2014;124:5109–28.
42. Federici C, Petrucci F, Caimi S, Cesolini A, Logozzi M, Borghi M, et al.
Exosome release and low pH belong to a framework of resistance of human
melanoma cells to cisplatin. PLoS ONE 2014;9:e88193.
43. Lv MM, Zhu XY, Chen WX, Zhong SL, Hu Q, Ma TF, et al. Exosomes mediate
drug resistance transfer in MCF-7 breast cancer cells and a probable
mechanism is delivery of P-glycoprotein. Tumour Biol 2014;35:10773–9.
44. Ma X, Chen Z, Hua D, He D, Wang L, Zhang P, et al. Essential role for TrpC5containing extracellular vesicles in breast cancer with chemotherapeutic
resistance. Proc Natl Acad Sci U S A 2014;111:6389–94.
45. Choi DY, You S, Jung JH, Lee JC, Rho JK, Lee KY, et al. Extracellular vesicles
shed from gefitinib-resistant nonsmall cell lung cancer regulate the tumor
microenvironment. Proteomics 2014;14:1845–56.
46. Boelens MC, Wu TJ, Nabet BY, Xu B, Qiu Y, Yoon T, et al. Exosome transfer
from stromal to breast cancer cells regulates therapy resistance pathways.
Cell 2014;159:499–513.
47. Bergsmedh A, Szeles A, Henriksson M, Bratt A, Folkman MJ, Spetz AL, et al.
Horizontal transfer of oncogenes by uptake of apoptotic bodies. Proc Natl
Acad Sci U S A 2001;98:6407–11.
48. Ehnfors J, Kost-Alimova M, Persson NL, Bergsmedh A, Castro J, LevchenkoTegnebratt T, et al. Horizontal transfer of tumor DNA to endothelial cells in
vivo. Cell Death Differ 2009;16:749–57.
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Published OnlineFirst March 31, 2016; DOI: 10.1158/0008-5472.CAN-15-2804
Implications of Extracellular Vesicle Transfer on Cellular
Heterogeneity in Cancer: What Are the Potential Clinical
Ramifications?
Anoek Zomer and Jacco van Rheenen
Cancer Res 2016;76:2071-2075. Published OnlineFirst March 31, 2016.
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