## 4 Studies of the Molecular Profiling of EVs as Potential Biomarkers
### Proteome Profiling Analysis of EVs in Multiple Cancers
\begin{table}
\begin{tabular}{c c c c} \hline
**Cancer Type** & **Biomarker** & **Sample** & **Authors** \\ \hline Thyroid Cancer & TLN1, ITGB2, SRC and CAPNS1 & serum derived-EVs & [4] \\ Breast Cancer & CD24 and EpCAM & serum derived-EVs & [84] \\ Breast Cancer & HER2 & plasma derived-EVs & [87] \\ Breast Cancer & GPC-1, ADAM10, GLUT-1 and desintegrin & _In vitro_: MDA-MB-231 and MCF-10A cell lines & [88] \\  & Del-1, 14-3-3 epsilon protein, \(\beta\)-actin, & _In vitro_: cell line MDA-MB-231 & [89] \\ Breast Cancer & annexin A1 / 5, heat shock protein 71, and galectin-binding protein 3 & _In vitro_: cell line MDA-MB-231 & [89] \\ Pancreatic Cancer & GPC-1 & serum derived-EVs & [90] \\ Colorectal Cancer & GPC-1 & plasma derived-EVs & [91] \\ Colorectal Cancer & CK19, TAG72, and CA125 & plasma derived-EVs, CRC cells, tumor & [92] \\ Ovarian Cancer & CA125 and HE4 & interstitial fluid & [93, 94] \\ Colon Cancer & CD147 & _In vitro_: HCT15 and HCT116 & [95] \\ Colon Cancer & TSPAN1 & _In vitro_: HCT-116 and HT-29 CC cell lines & [96] \\ Colon Cancer & annexin & plasma derived-EVs & [97] \\ Hepatic Cancer & AMPN, VNN1, plgR, FCN1 and NEP & serum derived-EVs & [98] \\ Hepatic Cancer & FIBG, A1 AG1 and S100A8 & serum derived-EVs & [98] \\  & FCN2, ITH4, FIBG; MUC1, EGFR, EpCAM, and others. & serum derived-EVs; EGI1, TFK1 cell & \\ Cholangiocarcinoma & FCN2, ITH4, FIBG; MUC1, EGFR, EpCAM, and others. & lines and non-tumor & [98] \\  & EpCAM, ASPR1, annexin V and taMPs & serum derived-EVs & [99] \\ Cholangiocarcinoma & fetuin-A and HSP90B & _In vitro_: M213 and M213D5 cell lines & [100] \\ Hepatic Cancer & CAP1 & _In vitro_: MHCC97-H and MHCC97-L cell lines & [101] \\ Gastric Cancer & PSMA3 and PSMA6 & serum derived-EVs & [102] \\ \hline \end{tabular}
\end{table} 
Table 1: Summary of the potential protein biomarkers identified in EVs from different cancer types. Another study revealed that glypican-1 (GPC-1), disintegrin and metalloproteinase domain-containing protein 10 (ADAM10), and glucose transporter 1 (GLUT-1) were upregulated in triple negative metastatic cancer cell line MDA-MB-231 (MDA) compared to the control immortalized epithelial breast tissue cell line MCF-10A (MCF) [88] (Table 1). GPC-1 is a cell surface proteoglycan protein involved in the control of cellular growth and differentiation by activation of mitogenic signaling by heparin-binding growth factors. It has been observed to be highly expressed in tissues from BC compared to healthy patients [88]. ADAM10, a transmembrane protease protein, is very abundant in high-grade tumors, and these levels correlated with negative outcomes for the basal subtypes of BC patients [113]. Lastly, GLUT-1 is a solute carrier protein that facilitates the transport of glucose across the plasma membranes. Its expression is correlated with high-grade BC cancer and increased proliferative activity, and its absence significantly increased disease-free survival in BC patients [114]. On the other hand, proteomic analyses from cell culture EVs have also provided other potential biomarker candidates in BC, including 14-3-3 epsilon protein, \(\beta\)-actin, annexin A1/5, heat shock protein 71, and galectin-binding protein 3 [89] (Table 1). In addition, Del-1 has been reported as an early-stage BC EVs biomarker [115]. Del-1 was first identified as an extracellular matrix protein having 3 N-terminal epidermal growth factor-like domains and the discoidin I-like or factor V C domains, C1 and C2 [116]. The striking decrease in Del-1 concentrations in plasma after surgery suggests that this protein could be a useful surveillance biomarker to assess the response of breast cancer patients to cancer therapies [115]. Protemos have also contributed to identifying potential biomarkers in other types of cancer, such as GPC-1, through showing increased serum EVs of pancreatic cancer (PC) patients [90] (Table 1), as well as in plasma from colorectal cancer (CRC) patients [91] (Table 1). All the studies dealing with the presence of GPC-1 in EVs confirm its potential role as a clinical cancer biomarker.