Extracellular Vesicles: Potential Applications in Cancer Diagnosis, Prognosis, and Epidemiology

Mukesh Verma; Tram Kim Lam; Elizabeth Hebert; Rao L Divi

BMC Clin Pathol. 2015;15(6)

Conclusions

Despite the challenges cited above, the scientific community is interested in these tissue-derived vesicles and their multiple roles and functions. EVs hold great promise for cancer diagnosis and treatment. Likewise, there are many potential uses of EVs in cancer research and epidemiology.

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Abbreviations
EGFR: Epidermal growth factor receptor, EpCAM: Epithelial cell adhesion molecule, ESCRT: Endosomal sorting complex required for transport, EVs: Evesicles, mRNA: messenger RNA, miRNA: microRNA, MVs: Microvesicles

Acknowledgements
We are thankful to Joanne Brodsky of Scientific Consulting group (SCG) for reading the manuscript and providing comments.

Table 1. Extracellular vesicles' analysis in different tumor types
Cancer types	Comments	References
Bladder cancer	Exosomes from urine contain the angiogenesis-promoting protein EDIL-3	[99]
Breast cancer	Microvesicle numbers and the amount of focal adhesion kinase and EGFR in plasma fractions were associated with different stages of breast cancer	[89]
Colorectal cancer	Proteomic analysis was conducted on EVs from colorectal cancer cells	[103]
Gastric cancer	Proteomic analysis was conducted on EVs from gastric cancer patients	[31]
Glioblastoma	Microvesicle RNA biomarkers of glioblastoma multiforme were identified	[29,55]
Head and neck cancer	Exosomes and microvesicles from patient saliva were used for diagnosis	[104]
Lung cancer	Proteins isolated from microvesicles in pleural effusions due to lung cancer were characterized to identify diagnostic markers	[3]
Melanoma	Proteomic analysis was conducted on exosomes from melanoma patients were used for	[105]
Ovarian cancer	Exosomes from ovarian cancer patients contain different sets of proteins and miRNAs compared to exosomes from normal subjects; the amount of circulating exosomes was 4 times higher in patients	[1,2,92,93,106]
Pancreatic cancer	EVs were used in diagnosing pancreatic cancer	[30]
Prostate cancer	Urine exosomes expressed higher levels of PCA-3 and TMPRSS2-ERG	[58]

Table 2. Biofluids/biospecimens used to isolate extracellular vesicles for cancer diagnosis, prognosis, and epidemiology
Biospecimen	Comments	References
Ascites	Used in diagnosing ovarian cancer and determining its aggressiveness	[39]
Blood and plasma	Used in diagnosing ovarian cancer and breast cancer	[89,93]
Breast milk	Exosomes were isolated from breast milk	[36]
Mesenchymal stem cells	Suppression of angiogenesis shown in tumor cells mediated by miR-16 isolated from exosomes	[5]
Pregnancy-associated sera	Exosomes were isolated in different stages of pregnancy	[107]
Saliva	Exosomes and MVs found in patient saliva	[104]
Stem cells	Renal stem cells contained MVs with angiogenesis-specific mRNAs and miRNAs	[94]
Tissues	Ovarian cancer tissues were used to isolate exosomes and then in isolating miRNAs from them	[1,14]
Urine	Urine was used to isolate exosomes and in analyzing proteins by MS or transcriptome analysis	[58,99]

Table 3. Methods for isolating extracellular vesicles (EVs)
EV Isolation Method	Sample	~ Time required for isolation	Reference/Company
Deferential centrifugation	Serum, urine and cell culture supernatant	2–15 h	[79]
Deferential centrifugation	Cell culture supernatant or saliva	2–15 h	[108,109]
Density gradient centrifugation	Cell culture supernatant	24 h	[80]
Sequential membrane filtration	Cell culture supernatant and biofluids	24 h	[81,110]
Nanomembrane	Urine	<2 h	[82]
Size exclusion chromatography	Plasma	20 min	[111]
Microfluidics	Cell culture supernatant	-	[112]
Nanoshearing	Cell culture supernatant and serum	-	[46]
ExoCap	Cell culture supernatant and biofluids	30 min	MBL International
ExomiR	Cell-free biofluids' exosomal RNA	20 min	Bioo Scientific
Exo-spin	Cell culture supernatant, urine and saliva	3 h	Cell Guidance Systems
ExoQuick	Serum, plasma, ascites, urine, CSF and cell culture supernatant	2–15 h	System Bioscinces
miRCURY	Serum, plasma, cells, urine and CSF	2 h	Exiqon
Total exosome isolation	Cell culture supernatant and biofluids	2 h	Life Technologies
PureExo	Serum, plasma and cell culture supernatant	2 h	101Bio
ME Kit based on Vn96 peptide binding to heat shock proteins on exosomes	Serum, plasma, urine, cell culture supernatant	30 min	New England Peptide
Streptavidin-biotin-specific antibody to a known antigen on exosome	Cell culture supernatant	>12 h	Life Technologies
Anti-tetraspanin antibody-magnetic bead based	Cell culture supernatant	>12 h	Life Technologies
Anti-EpCAM-antibody magnetic bead based	Cell culture supernatant	>4 h	Life Technologies