Table 5.
Minimal Information for Studies of EVs (MISEV) guidelines related information about EV isolation and characterization in the studies discussed in this review, investigating human- and mouse-derived EVs.
| Patient studies | |||||||||
|---|---|---|---|---|---|---|---|---|---|
| Patient info (disease/controls) | Biofluid/tissue as EV source | EV cellular source (marker) | Complement content EVs | EV isolation | EV characterization | Ref. | |||
| Global quantification | Protein marker detection | ||||||||
| Dementia | FTLD (genetic GRN/C9orf72, sporadic) | HC | Plasma | Bulk | ↑ C1q, C3, C4 cargo per EV in FTLD ↓ C4 cargo in GRN+(homo) vs GRN+(het) and sporadic ↑ C4 cargo in sporadic vs GRN+(het) ↑ C4 EV/plasma ratio in sporadic vs GRN+ FTLD ↓ C4 EV/plasma ratio in GRN+ (homo) vs HC and other FTLD groups ↑ C3 EV/plasma ratio in sporadic vs HC and other FTLD groups |
Total Exosome Isolation kit | ☐ A ☒ B (NTA) ☐ C ☐ D ☐ E |
☒ 1 (CD9) ☒ 2 (TSG101) ☐ 3 ☒ 4 (Calnexin) |
(136) |
| AD, FTLD | A/G HC | Plasma | Bulk (CD81+) Astrocytes (GLAST+) Neurons (L1CAM+) |
AEVs and NEVs of AD patients are neurotoxic (induction Membrane Attack Complex (MAC) expression, membrane disruption, reduced neurite density, decreased cell viability) | ExoQuick + IP | ☒ A (0,5 ml) ☒ B (NTA) ☒ C (Bradford) ☐ D ☒ E |
☒ 1 (CD81, CD9, CD63) ☒ 2 (ALIX) ☒ 3 (ApoA1) ☒ 4 (GM130) |
(137) | |
| AD (mild) AD (moderate) |
A/G HC Pre-clinical AD |
Plasma | Astrocytes (GLAST+) |
↑ C1q, C4b, C3d, factor B, factor D, fBb, C3b and TCC (C5b-C9) in AEVs of mild AD vs HC. Mean complement levels higher in moderate AD vs preclinical AD. |
ExoQuick + IP | ☒ A (250 µl) ☐ B ☐ C ☐ D ☐ E |
☒ 1 (CD81, CD59, CD55) ☐ 2 ☐ 3 ☐ 4 |
(138) | |
| MCIC AD (mild, moderate) |
MCIS A/G HC |
Plasma | Astrocytes (GLAST+) |
↑ C1q, C4b, fD, fragment Bb, C5b, C3b, C5b-C9 in AEVs of MCIC vs MCIS. ↓ CD46, CD59, and type 1 complement receptor in AEVs in MCIC vs MCIS. |
ExoQuick + IP | ☒ A (250 µl) ☐ B ☐ C ☐ D ☐ E |
☒ 1 (CD81, CD59, CD55) ☐ 2 ☐ 3 ☐ 4 |
(139) | |
| AD Braak stage V-VI |
A/G HC | Brain tissue (parietal cortex) | Microglia (CD11b+) |
Proteomic EV analysis ↑ C4, CD59 in MEVs from AD vs HC |
GentleMACS tissue dissociation Sucrose density gradient UC + IP |
☐ A ☒ B (TRPS) ☐ C ☒ D (lipidomics) ☒ E |
☒ 1 (CD9, CD81, CD63, CD11b) ☒ 2 (syntenin-1) ☒ 3 ☒ 4 (GM130, Calnexin) |
(140) | |
| Parkinson’s disease | PD HY stages II and III |
HC | Plasma | Bulk | Proteomic EV analysis ↓ Clusterin, C1r in PD vs HC |
SEC (EV-Second) | ☒ A (200 µl) ☐ B ☒ C (Bradford) ☐ D ☒ E |
☒ 1 (CD81, CD9) ☐ 2 ☐ 3 ☐ 4 |
(141) |
| PD Mild and severe |
HC | Serum | Bulk | Proteomic EV analysis ↓ C1q in PD vs HC ↑ Clusterin, C1r in progression from mild to severe PD ↓ C1q in progression from mild to severe PD |
DUC | ☒ A (5 ml) ☐ B ☒ C (BCA) ☐ D ☐ E |
☐ 1 ☐ 2 ☐ 3 ☐ 4 |
(142) | |
| Multiple sclerosis | RRMS, pMS | HC | Plasma | Astrocytes (GLAST+) |
↑ C1q, C3b/iC3b, C5, C5a, fH (pMS vs. HC) ↑ C1q, C3, C3b/iC3b, C5, C5a, fH (RRMS vs HC) No difference: C4, C9, fB (MS vs. HC) |
ExoQuick + IP | ☒ A (0,5 ml) ☒ B (NTA) ☐ C ☐ D ☐ E |
☐ 1 ☐ 2 ☐ 3 ☐ 4 |
(143) |
| RRMS | IIH | CSF | Bulk | Proteomics on CSF-EVs C3b, C4, C6, fB, fH uniquely enriched in RRMS-EVs vs RRMS-CSF, while not enriched in IIH. |
Precipitation + SEC (ExoSpin) | ☒ A (5 ml) ☒ B (NTA) ☒ C (NanoDrop) ☐ D ☒ E |
☒ 1 (CD81, CD9) ☒ 2 (TSG101) ☒ 3 (HSA) ☐ 4 |
(144) | |
| MS | NA | CSF | NA | MAC-containing “vesicles” | NA | ☐ A ☐ B ☐ C ☐ D ☐ E |
☐ 1 ☐ 2 ☐ 3 ☐ 4 |
(145) | |
| Ischemic stroke | Symptomatic IS | HC | Serum | Bulk | Proteomic EV analysis ↑ C1qB, C1r in IS vs HC |
ExoTrap | ☒ A (500 µl) ☐ B ☒ C (BCA) ☐ D ☐ E |
☒ 1 (CD9, CD81) ☒ 2 (TSG101) ☐ 3 ☒ 4 (Calnexin) |
(146) |
| IS (CS, CSC) |
HC | Serum | Bulk Neurons (L1CAM+) |
Proteomic EV analysis C1qA: unique for serum EVs and NEVs from CSC-IS patients. C3 abundant in IS (CSC and CS) and HC in serum EVs and NEVs. |
ExoQuick Ultra (Bulk) SmartSEC + IP (NEVs) |
☐ A ☒ B (NTA) ☐ C ☐ D ☒ E |
☒ 1 (CD63, CD81) ☒ 2 (ALIX) ☒ 3 (Albumin) ☐ 4 |
(147) | |
| Brain tumor | GBM | HC | Plasma | Bulk | Proteomic EV analysis ↑ C3, C5, C1q, fH in GBM vs HC |
DUC | ☒ A (5 ml) ☒ B (NTA) ☒ C (MicroBCA) ☐ D ☒ E |
☒ 1 (CD81, CD9, CD63, CD41a) ☒ 2 (HSP70) ☐ 3 ☒ 4 (GM130) |
(148) |
| GBM | HC | Plasma | Bulk | Proteomic EV analysis ↑ C3, C4b in GBM vs HC |
DUC | ☒ A (15 ml blood) ☒ B (NTA) ☐ C ☐ D ☒ E |
☒ 1 (CD9, CD63) ☒ 2 (TSG101) ☐ 3 ☐ 4 |
(149) | |
| GBM Grade II-IV |
MEN HC |
Plasma | Bulk | Proteomic EV analysis ↑ C3 in GBM vs controls |
SEC (qEV, Izon) | ☒ A (0.5 ml) ☒ B (NTA) ☐ C ☐ D ☒ E |
☒ 1 (CD9, ITGA2B, ITGA6, PDCD6IP) ☒ 2 (ANXA1/2/6, FLOT1, HSP90A1B, GAPDH, HIST1H4A) ☐ 3 ☒ 4 |
(150) | |
| Mouse studies/primary culture studies | |||||||||
| Mouse model Cell culture |
Biofluid/tissue as EV source | EV cellular source (marker) | Complement content EVs | EV isolation | EV characterization | Ref. | |||
| Global quantification | Protein marker detection | ||||||||
| Dementia | AβO CPE primary culture |
Scram-bled CPE primary culture |
Culture medium | CPE cells | Proteomic EV analysis ↑ C3 in AβO-stimulated CPE vs scrambled |
SEC (qEV) | ☒ A (500 µl) ☒ B (NTA) ☐ C ☐ D ☒ E |
☒ 1 (CD81, CD9) ☒ 2 (TSG101) ☐ 3 ☒ 4 (Calnexin) |
(151) |
| 2xTg-AD 5xFAD 3xTg-AD |
WT | Plasma | Neuron (L1CAM+) Astrocyte (GLAST+) |
↑ AEV C1q in 3xTg-AD vs WT | IP (ExoSORT) | ☐ A ☒ B (NTA) ☒ C (Bradford) ☐ D ☐ E |
☒ 1 (CD81, CD9, CD63) ☒ 2 (FLOT1) ☒ 3 (APOA, albumin) ☒ 4 (Calnexin) |
(152) | |
| CAST.APP/PS1 | CAST WT HC | Brain tissue | Bulk | Proteomic EV analysis C1qa, C1qc increased in EVs from CAST APP/PS1 vs WT, but not significant |
DUC + sucrose gradient | ☒ A (0.4g) ☒ B (NTA) ☒ C (BCA) ☐ D ☒ E |
☒ 1 (CD9, CD81, CD63, ITGA) ☒ 2 (ANXA5) ☐ 3 ☒ 4 (GM130, CYC1) |
(153) | |
| Brain tumor | GBM mouse model Longitudinal samplings (baseline, T1, T2) |
NA | Serum | Bulk | Proteomic EV analysis Complements among group of deregulated proteins in EVs C1ra and C1sa deregulated in EVs between T2 and T1 stages. |
Precipitation (Total Exosome Isolation reagent) or SEC | ☒ A (50µl) ☒ B (DLS) ☐ C ☐ D ☒ E |
☒ 1 (CD9, ITGA) ☒ 2 (ANXA4, ANXA5, ANXA7) ☐ 3 ☐ 4 |
(154) |
| GBM mouse model Longitudinal samplings (baseline, pre-symptomatic T1, symptomatic T2) |
NA | Serum | Bulk | Proteomic EV analysis ↓ C4b in T1 vs baseline ↑ C1qa, C1ra, C1s1 in T1 and T2 vs baseline |
SEC (qEV – 70nm) | ☒ A (50µl) ☐ B ☒ C (MicroBCA) ☐ D ☐ E |
☒ 1 (ITGB1) ☒ 2 (HSPA8, ACT) ☒ 3 (Albumin) ☐ 4 |
(155) | |
For global EV quantification requirements, consult checkboxes A-E. Checkbox A) Cell number/fluid volume/tissue mass from which EVs were isolated. Checkbox B) Analysis of particle number. Checkbox C) Analysis of protein amount. Checkbox D) Analysis of lipid amount. Checkbox E) Analysis by electron microscopy. For information regarding EV protein marker detection that has been conducted by the indicated studies, consult checkboxes 1-4. Checkbox 1) Transmembrane or Glycosylphosphatidylinositol (GPI)-anchored protein(s) localized in cells at plasma membrane or endosomes. Checkbox 2) Cytosolic protein(s) with membrane-binding or -association capacity. Checkbox 3) Assessment of presence/absence of expected contaminants. Checkbox 4) For small EVs <200nm: verifying protein(s) associated with compartments other than plasma membrane or endosomes. extracellular vesicles (EVs), astrocyte-EV (AEV), neuronal-EV (NEV), microglial-EV (MEV), not described (ND), not applicable (NA), idiopathic intracranial hypertension (IIH); multiple sclerosis (MS), relapsing-remitting MS, progressive MS (pMS), Alzheimer’s disease (AD), Fronto-Temporal Lobar Degeneration (FTLD), Parkinson’s disease (PD), Glioblastoma multiforme (GBM), meningioma (MEN), ischemic stroke (IS), subcortical (SC), cortical-subcortical (CSC), mild cognitive impairment (MCI), MCI stable (MCIS), MCI converting to dementia (MCIC), Hoehn and Yahr (HY), healthy controls (HC), age-gender matched healthy controls (A/G HC), homozygous (hom), heterozygous (het), Nanoparticle Tracking Analysis (NTA), Bicinchoninic Acid Assay (BCA) Bradford assay (BA), mass spectrometry (MS), differential ultracentrifugation (DUC), ultracentrifugation (UC), Annexin A5 (ANXA5), flow cytometry (FC), transmission electron microscopy (TEM), size exclusion chromatography (SEC), high sensitivity flow cytometry (hsFC), immunoprecipitation (IP), Tunable Resistive Pulse Sensing (TRPS), factor H (fH), factor B (fB), factor D (fD), terminal complement complex (TCC), human serum albumin (HSA), heat-shock protein (HSP), A-beta oligomers (AβO), choroid plexus epithelial cells (CPE), wildtype (WT).