Table 2. Clinical application of EV markers for PC.
Abbreviations: AC: ampullary carcinoma; AEP: asparaginyl endopeptidase; AUC: area under curve; bPaTu: benign pancreatic tumor; BPD: benign pancreatic disease; BPT: benign pancreatic tumors; ELISA: enzyme-linked immunosorbent assay; ddPCR: droplet digital polymerase chain reaction; DFS: disease-free survival rates; ExoTENPO: Exosome Track-Etched Magnetic Nanopore; IPMN: intraductal papillary mucinous neoplasm; LOD: limit of detection; LSPR: localized surface plasmon resonance; MAF: mutant allele frequency; MIF: macrophage migration inhibitory factor; NED: no evidence of disease post-diagnosis; NC: normal control; nPES: nanoplasmon-enhanced scattering; OS: overall survival; PC: Pancreatic Cancer; PCPL: pancreatic cancer precursor lesions; PDAC: pancreatic ductal adenocarcinoma; Pt: chronic pancreatitis; POD: progression of disease post-diagnosis; PaCIC: pancreatic cancer initiating cells; SCA: serous cystadenoma; UC: ultracentrifugation.
| EV markers for pancreatic cancer | EV Isolation | EV marker analysis | Analytical performance | Clinical Utility | Sample size | Clinical performance /Statistical analysis | Blinded? Yes/No | Sample type/single site collection? | Ref |
|---|---|---|---|---|---|---|---|---|---|
| Glypican-1 | UC | Flow cytometry | Not specified | Early detection | Discovery Cohort: NC: N=100, BPD: N=26; PCPL: N=5; PDAC: N=190, Validation Cohort:, NC: N=20; BPD: N=6; PDAC: N=56 | PC vs other groups AUC: 1 Sensitivity: 100% Specificity: 100% |
Yes | Serum/ discovery cohort yes; validation cohort yes | 38 |
| Metastatic disease burden | No metastases: N=18, Lymph node metastases: N=134, Distant metastases: N=32 | Distant metastases vs No metastases or Lymph node metastases statistically significant | Yes | Serum/not specified | |||||
| Monitor surgery responses | BPD: N=4, PCPL: N=4, PDAC: N=29 | Pre vs post resection (PDAC and PCPL statistically significant) | |||||||
| Prognosis | PDAC: N=29 | Improved OS and DFS if drop of exosomal GPC1+ >= median | |||||||
| Glypican-1 | UC | Tandem mass spectrometry | Accuracy Precision Sensitivity Matrix effect Spike recovery Sample Stability |
Diagnosis | NC: N= 6, Pt: N = 3, PDAC: N = 3 | Not statistically significant | Not specified | EDTA Plasma/ not specified | 39 |
| Monitor surgery responses | Matched pre- and post- surgical resection: N=3 | Not statistically significant | |||||||
| Glypican-1 | UC | ELISA | Not specified | Diagnosis | BPD: N=16, IPMN: N=7 Pt: N=6, SCA: N=3 PDAC: N=27 |
No significant difference AUC= 0.59, Sensitivity=74%specificity=44% for detecting PDAC. | Not specified | EDTA Plasma /Yes | 40 |
| Monitor surgery responses | Matched pre- and post- surgical resection: N=11 | Statistically significant | |||||||
| Glypican-1 and CD63 | None | AC electrokinetic microarray chip | Not specified | Diagnosis | NC: N=11, BPD: N=7, PDAC: N=20 | PDAC vs other groups, AUC: 0.79, Sensitivity: 81%, Specificity: 70% | Yes | Serum/Yes | 41 |
| Glypican-1 | Spin column-based method | Flow cytometry | Not specified | Diagnosis | NC: N=16, PC: N=28 | Statistically significant | Not specified | Plasma ((anticoagulant not specify)/Yes | 42 |
| Prognosis | PC: N=28 | Improved OS with great decrease of Glypican-1+ EV | |||||||
| EphA2-EV | Isolation free | nPES | Cutoff, Precision LOD, Linearity | Early detection | Discovery Cohort: NC: n = 10, Pt: n = 10, PC: n = 10 Validation Cohort: NC: n = 48, Pt: n = 48, PC: n = 49 (8 stage I, 29 stage II, and 12 stage III) | AUC: 0.93–0.96 Sensitivity: 86%−94% Specificity: 85% |
No | Plasma (anticoagulant not specify)/yes | 43 |
| Not specified | Monitor treatment responses | good/partial responses: N= 13 poor responses: N= 10 |
good/partial vs poor responses: statistically significant | No | Plasma (anticoagulant not specify) /yes | ||||
| EphA2 | UC | ELISA | LOD Linearity | Diagnosis | NC: n = 10, Pt: n = 10, PC: n = 10 | PC vs other groups Statistically significant |
No | Plasma (anticoagulant not specify) /yes | |
| Integrin αv | UC | ELISA | Not specified | Prognosis | NC: N=13, NED: N=14, POD to liver: N=13 | NED vs POD to liver, Statistically significant | Not specified | Plasma (anticoagulant not specify)or serum/No | 44 |
| MIF | UC | ELISA | Not specified | Prognosis | NC: N=15, NED= 10, POD N=12, POD to liver: N=18 | NED vs POD Statistically significant |
No | EDTA plasma/No | 45 |
| AEP | ExoQuick | Western blot | Not specified | Exploratory | PDAC: N=3,Pt: N=3 | Statistically significant | N/A | Serum/Yes | 46 |
| CD44v6, Tspan8, EpCAM, MET, CD104, miR‐1246, miR‐4644, miR‐3976 and miR‐4306 | UC | Flow cytometry, RT-qPCR | Cutoff | Diagnosis | Discovery cohort: NC: N=6, Non-PC cancer:
N=3, Pt: N=7, PC: N=37 bPaTu: N=5, Validation cohort: NC: N=12,Non-PC cancer: N=8, Pt: N=16, PC: N=75, bPaTu: N=17 |
Combining all markers PC vs non-PC sensitivity: 100% specificity: 80% |
Yes | Serum/No | 47 |
| miR-17–5p, miR-21 | UC | RT-qPCR | Not specified | Diagnosis | NC: N=8, Pt: N=6, AC: N=6, PC: N=22, BPT: N=7 | PC vs non-PC, AUC: 0.887 and 0.897, Sensitivity: 92.6% and 81.5%, Specificity: 72.7% and 95.5% | Not specified | Serum/Yes | 48 |
| miR-10b | UC | LSPR-based sensor | LOD, Method
comparison Linearity |
Exploratory | NC: N=3, Pt: N=3, PDAC: N=3 | Statistically significant | Yes | Trisodium citrate Plasma/Not specified | 49 |
| miR-1246 and miR-4644 | Total Exosome Isolation Reagent | RT-qPCR | Threshold cycle values Cutoff | Exploratory | PC: N=12 NC: N=13 |
AUC: 0.763–0.833 Sensitivity: 0.667–0.833 Specificity: 0.769–1 |
Not specified | Saliva/Yes | 50 |
| miR-196a and miR-1246 | Exoquick | RT-qPCR | Not specified | Early detection | NC: N=15; PC stage I or IIA: N=15 | AUC: 0.73–0.81 | Not specified | Plasma (anticoagulant not specify)/No | 51 |
| miR-23b-3p | UC | RT-qPCR | Not specified | Diagnosis | NC: N=20, Pt: N=18, PC: N=16 | PC vs NC and Pt, Statistically significant | Not specified | Serum/Yes | 52 |
| miR-10b, −21, −30c, −106b, −20a, −181a, −483, -let7a, and −122 | UC | RT-qPCR | Cutoff | Diagnosis | NC: N= 6, Pt: N = 11 PDAC: N = 29 |
PDAC vs Normal, AUC: 0.57–1, Sensitivity: 62%−100%, Specificity: 100% | Not specified | EDTA Plasma/Not specified |
39 |
| miR-301a-3p | Total exosomes isolation reagent | RT-qPCR | Not specified | Diagnosis | PC: N=50, NC: N=12 | Statistically significant | Not specified | Serum/No | 53 |
| Prognosis | PC: N=50, miR-301a-3p low: N=20; high: N=30 | higher miR-301a expression was associated with poor OS | |||||||
| miR-122–5p, miR-193b-3p | ExoQuick | RT-qPCR | Not specified | Diagnosis | PC: N=31 NC: N=37 |
AUC=0.849 | Not specified | EDTA plasma/yes | 54 |
| miR-451a | UC | RT-qPCR | Not specified | Early detection | PC: Stage I N=7, Stage II N=43, NC: N=20 | Statistically significant | Not specified | Plasma (anticoagulant not specify)/Yes | 55 |
| Prognosis | high-miR-451a expression group:
N=25; |
The high miR-451a group showed a
significantly worse OS (P = 0.001) and DFS Significantly shorter OS and DFS in the high-miR-451a group |
|||||||
| miR-191, miR-21 and miR-451a | ExoQuick | RT-qPCR | Not specified | Early detection | PC: N= 32, IPMN: N= 29, Control: N= 22 | AUC: 0.768–0.862 Sensitivity: 69.6%−86.4% Specificity: 79%*81% |
Not specified | Serum/Yes | 56 |
| miR-21 | cutoff | Prognosis | Not specified | Significantly shorter OS in the high-miR-21 group | |||||
| miR-21 and miR-451a | Not specified | Monitor treatment responses | Not specified | Statistically significant | |||||
| Sox2ot | ExoQuick | Microarray and qRT-PCR | Not specified | Prognosis | High Sox2ot expression group,
N=25; Low Sox2ot expression group, N=31 |
patients with high exosomal Sox2ot expression had lower overall survival rates | Not specified | Plasma (anticoagulant not specify)/Yes | 57 |
| Monitor surgery responses | Matched pre- and post- surgical resection: N=16 | Statistically significant | |||||||
| Mutant KRAS and TP53 DNA | UC | ddPCR | Threshold (intensity) limit for mutant alleles | Exploratory | PDAC: N=48 IPMN: N = 7, Pt: N = 9, Others:
N = 12, NC: N= 114 |
PDAC
KRASG12D:
39.6% TP53R273H : 4.2% |
Not specified | Serum/No | 58 |
| KRAS MAF | UC | ddPCR | Cutoff | Prognosis | Localized PDAC: N=13 | Longer DFS if exoKRAS MAF was <1% | Yes | Sodium Heparin plasma/not specify | 59 |
| KRAS MAF | UC vs UC+ immunocapture pulldown | ddPCR | Not specified | Exploratory | PDAC No-pulldown: N=136 Pulldown: N=37 |
Significantly higher KRAS MAF in the pulldown-cohort | Not specified | Acid Citrate Dextrose Plasma/Not specified | 60 |
| mRNA (ARG1, CK18, CD63, Erbb3, KRAS, GAPDH, H3F3A, ODC1) | EpCAM-based isolation on the ExoTENPO | RT-qPCR | Not specified | Exploratory | Training cohort, NC: N = 5, PC: N = 5, Validation cohort, NC: N = 12, PC: N = 12 | Sensitivity: 100% Specificity: 100% |
Yes for validation | EDTA plasma, Streck Cell-Free DNA BCT, or serum/Yes | 61 |