Table 1.
Main results of the systematic review of the studies investigating the potential applications of liquid biopsy and liquid biopsy-based biomarkers in MPNs.
| Author and Year | Study Location | Study Design | MPN | Patients No., Time Point | Parameters Assessed | Methods of Evaluation | Main Results |
|---|---|---|---|---|---|---|---|
| Garcia-Gisbert et al. (2020) [15] | Spain | Cross-sectional | PV, ET, PMF | 107 diagnosis follow-up |
cfDNA | DNA isolation | ↑ cfDNA ↑ cfDNA, cfDNA/WBCs in PMF vs. PV, ET ↑ cfDNA in MPNs with thrombosis at diagnosis/during follow-up ↑ VAF JAK2, MPL, SRSF2 in cfDNA vs. granulocyte DNA |
| Barone et al. (2019) [16] | Italy | Cross-sectional | ET, PMF, SMF | 81 follow-up |
PLTMVs, MKMVs, EMVs, MMVs, | Flow cytometry | ↓ MKMVs in JAK2(+)/CALR(+)/TN MF ↓ MKMVs, ↑ PLTMVs in MF & ET ↑ PLTMVs in ET vs. MF ↑ PLTMVs in JAK2(+)/CALR(+) MF vs. TN MPNs or controls ↓ PLTMVs in TN vs. JAK2(+)/CALR(+) MF ↓ MKMVs, ↑ PLTMVs in JAK2(+)/CALR(+) ET vs. controls & TN-ET ↑ PLTMVs, ↓ MKMVs in high/intermediate 2 vs. low/intermediate-1 risk MF & controls MKMVs in MF: (+) correlation wit PLTs, (−) with IL-6 PLTMVs in MF: (−) correlation with splenomegaly degree, (+) with P-selectin, thrombopoietin ↓ PLTMVs, ↑MKMVs in RUX spleen-responders in MF at baseline MKMVs <19.95% = spleen non-responders RUX ↓PLTMVs, ↑MKMVs in spleen-responders at 6 months ↑EMVs, MMVs in MF RUX ↓ EMVs in spleen-responders |
| Barone et al. (2020) [17] | Italy | Experimental | MF | 30 diagnosis |
MEVs-CK | Flow cytometry | RUX ↑ IL-1β, IL-6, TNF-α in LPS-stimulated MF monocytes |
| Poisson et al. (2020) [18] | France | Experimental | JAK2V617F(+) MPNs | 7 follow-up |
Plasma MVs | Flow cytometry | MVs from JAK2 V617F-positive MPNs ↑phenylephrine-induced contraction in mice aorta |
| Pecci et al. (2015) [19] | Italy | Cross-sectional | PV, ET, PMF | 5 follow-up |
PaCS, proteasome levels | EM+IGA, WB, ELISA |
↑ PaCS in PLTs, granulocytes ↑ proteasome levels in PLTs and granulocytes extracts ↑ proteasome levels in plasma |
| Caivano et al. (2015) [20] | Italy | Cross-sectional | PMF | 5 follow-up |
MVs | Flow cytometry | ↑ small-diameter MVs |
| Fel et al. (2019) [21] | Germany | Case-control | PV | 9 follow-up |
EVs | Liquid chromatography, tandem mass spectrometry | ↑ CD42d+, CD71+, CD62L+ cells in PV ↑ APs, ↑ inflammatory/immune/angiogenic/procoagulant markers ↑ 13x transferrin receptor protein 1 ↑ 11.2x heparanase ↑ 5–6x plasminogen activator inhibitor 1, histone H4 and H2B, angiogenin ↑ 4–5x matrix metalloproteinase-9, neurogenic locus notch homolog protein 3, lysozyme C, histone H3, L-selectin, lactotransferrin, solute carrier family 2 ↑ 3–4x coagulation factor XI, myeloperoxidase, C-reactive protein, vinculin, platelet multimerin-1 |
| Forte et al. (2011) [22] | Italy | Cross-sectional | PMF | 29 follow-up |
EVs | Flow cytometry | ↓ MK-EVs in JAK2 V617F (+) & TN-MF ↓ PLT-EVs in TN-MF, controls vs. JAK2 V617F (+) MF ↑ in vitro survival, ↑ miR-361-5p of TN-MF EVs miR-34a-5p, miR-222-3p, miR-361-5p upregulated in JAK2 V617F (+) & TN-MF miR-127-3p upregulated in JAK2 V617F (+) MF (+) miR-34a-5p, (−) miR-212-3p & JAK2 V617F VAF association |
| Zhang et al. (2017) [23] | China | Cross-sectional | PV, ET, PMF | 92 follow-up |
PMPs, EMPs, RMPs, TF+MPs | Flow cytometry | ↑ RMPs, ↑ PMPs, ↑ EMPs, ↑ TF+MPs PMF vs. PV: ↑ RMPs, ↑ PMPs, ↑ EMPs, ↑ TF+MPs ET vs. PV: ↑ EMPs |
| Villmow et al. (2003) [24] | Germany | Cross-sectional | PV, ET, PMF | 37 follow-up |
PMPs, PMAs, APs, PNCs, PMCs | Flow cytometry | ↑ APs ↑ PMPs in PV, ET, MF vs. CML, controls ↑ PNCs, ↑PMCs in ET, PV vs. MF, CML, controls |
| Trappenburg et al. (2009) [25] | Italy | Cross-sectional | ET | 21 follow-up |
PMPs, EMPs, GMPs, MMPs, TF+MPs | Flow cytometry | ↑ MPs, ↑ CD61(+) PMPs, ↓ CD63(+) PMPs, ↑ vWF, ↑ TF+MPs ↑ EMPs, i.e., CD62E(+), CD144(+) ↑ GMPs, i.e., CD66b(+) and CD66acde(+) ↑ MMPs, i.e., CD14(+) ↑ CD62E(+)/CD41(+) EMPs in ET with ↑ CV risk |
| Taniguchi et al. (2017) [26] | Japan | Cross-sectional | PV, ET, PMF, SMF | 59 follow-up |
PMPs, EMPs, TF+MPs | Flow cytometry | cytoreduction ↓ procoagulant, annexin V(+) MPs, ↓ TF+MPs anticoagulation ↓ MPs in MPNs 70% of annexin V(+) MPs = PMPs, i.e., CD41a(+) 30% of annexin V(+) MPs = EMPs, i.e., CD146(+), or CD45(+), i.e., leukocyte-derived history of thrombosis +/− no cytoreduction =↑ TF+MPs >84.7 TF+MPs/µL, documented CV risk = predictors of thrombosis in MPNs |
| Tan et al. (2013) [27] | China | Cross-sectional | PV | 23 follow-up |
PMPs, GMPs, EMPs, RMPs, TF+MPs | Flow cytometry | ↑MPs, ↑ PMPs, ↑ RMPs, ↑ GMPs, ↑ EMPs in PV vs. SP or controls ↑ PS(+) PLTs, RBCs in PV vs. SP or controls ↓ clotting time, ↑thrombin and FXase generation in PV HU ↓ MPs, ↑ PMPs, ↑ RMPs, PS(+) PLTs/RBCs in PV |
| Piccin et al. (2017) [28] | Italy | Cross-sectional | JAK2V617F(+) ET | 66 follow-up |
PMPs, EMPs, RMPs, TF+MPs | Flow cytometry | ↓ MPs, ↓ PMPs, ↑ NO, ↑ ADM in ET on ASA, HU+ASA ↑ EMPs, ↑ RMPs in untreated ET ↑ EMPs in ET vs. controls ↓ EMPs, ↓ ED-1 in ET on ASA+ANA |
| Ahadon et al. (2018) [29] | Malaysia | Case-control | PV | 15 diagnosis |
PMPs, EMPs | Flow cytometry | ↑ PMPs |
| Aswad et al. (2019) [30] | Czech Republic | Cross-sectional | PV, ET, PMF | 179 diagnosis follow-up |
PMPs, RMPs | Flow cytometry, functional coagulation assays | ↑ PMPs, RMPs ↑ PMPs in PV, ET vs. PMF ↑ procoagulant activity of MPs association of PMPs procoagulant activity and PMPs levels ↓ PMPs in MPNs with (+) history of thrombosis ↑ PMPs in JAK2 V617F (+) MPNs PMPs correlated with Hb, Ht, RBCs, PLTs, WBCs |
| Charpentier et al. (2016) [31] | France | Cross-sectional | ET | 74 diagnosis |
PMPs, RMPs, MMPs, GMPs, EMPs | Flow cytometry | ↑ total MPs, RMPs, PMPs in JAK2 V617F (+) vs. CALR(+)/TN-ET (+) associations of MPs with thrombin generation, phospholipid-dependent procoagulant activity ↑ procoagulant activity in JAK2 V617F (+) vs. CALR(+)/TN-ET ↑ MPs in high vs. intermediate/low thrombotic risk ET >4600 MPs/µL = high-risk of thrombosis in ET |
| Marchetti et al. (2014) [32] | Italy | Cross-sectional | ET | 73 follow-up |
MPs | Flow cytometry | ↑ ETP, ↑ peak of thrombin, ↓ lag-time, ↓ time to peak, ↓ clotting times ↑ ETP, ↑ peak of thrombin, ↓ lag-time, ↓time to peak in JAK2 V617F (+) vs. (−) ↓ clotting times in JAK2 V617F (+) vs. (−) JAK2 V617F predicts shortened clotting times (+) association of PCA and lag-time, time to peak (−) association of PCA and peak of thrombin, ETP removal of MPs ↓ EDT, ↑ clotting times in ET, controls ↑ TF, ↑ FVIIa/AT in JAK2 V617F (+) ET |
| Moles-Moreau et al. (2009) [33] | France | Cross-sectional | ET | 37 diagnosis |
PMPs | Flow cytometry | ↑ PMPs, PMPs/PLTs, ↑ CD36+ cells ↑ PMPs/PLTs ratio, ↑ CD36+ cells in ET vs. RT ↑ PMPs, PMPs/PLTs ratio in RT vs. controls |
| Connor et al. (2013) [34] | Australia | Cross-sectional | ET | 10 follow-up |
PMPs | Flow cytometry | ↑PMPs |
| Duchemin et al. (2010) [35] | France | Cross-sectional | PV, ET | 44 follow-up |
MPs | Functional assays | ↑ CPA, ↓ ETP in MPNs pre-/post-filtration of MPs ↑ CPA in JAK2 V617F (+), homozygous genotype ↑ TM-resistance, ↓free protein S (+) association of CPA and neutrophils, RBCs, PLTs (−) association of ETP and JAK2 V617F allele burden ↑ CPA, ↓ ETP in JAK2 V617F homozygous genotype MPNs post-filtration of MPs |
| Kissova et al. (2015) [36] | Czech Republic | Cross-sectional | PV, ET, PMF | 126 follow-up |
MPs | Flow cytometry | ↑ MPs ↑ PCA of MPs in PV vs. ET/ PMF ↑ PCA of MPs in JAK2 V617F (+) vs. (−) MPNs (+) association of PCA of MPs with Hb, Ht in PV association of PCA with PLTs ↑ PCA of MPs in MPNs with venous thrombosis history cytoreduction ↓PCA of MPs |
| Alonci et al. (2008) [37] | Italy | Cross-sectional | PV, ET, PMF | 40 follow-up |
CECs | Flow cytometry | ↑ CD34+ CECs, ↑ CD34+ CD133- VEGFR2+ CECs ↑ CD34+ CECs in PMF vs. ET, PV ↑ CD34+ CD133+ VEGFR2+ CECs in PMF, PV vs. controls, ET ↑ CD34+ CD133- VEGFR2+ CECs in PMF vs. PV |
| Belotti et al. (2011) [38] | Italy | Cross-sectional | ET | 39 follow-up |
CECs | Flow cytometry | ↑ CD146+ CD45-CECs, soluble E-selectin |
| Torres et al. (2013) [39] | Portugal | Cross-sectional | PV, ET | 17 follow-up |
CECs | Flow cytometry | ↑ MPNs, VTE vs. controls ↓ progenitor CECs in VTE vs. MPNs, controls ↑ CD62E+ CECs in MPNs vs. controls ↑ CD62E+, CD54+, CD142+ CECs in VTE vs. controls (+) associations of WBCs with total CECs, progenitor CECs, CD62E+CECs (−) associations of PLTs with CD54+CECs |
| Trelinski et al. (2010) [40] | Poland | Cross-sectional | ET | 65 diagnosis follow-up |
CECs | Flow cytometry | ↑total, activated, resting, progenitor, CD46+, apoptotic CECs, VEGF, soluble VEFGR 1 ↓ placenta growth factor |
| Trelinski et al. (2010) [41] | Poland | Cross-sectional | PV, ET | 46 follow-up |
CECs | Flow cytometry | ↑ total, activated, progenitor, apoptotic CECs ↑ resting CECs in ET versus PV, controls ↑ apoptotic CECs in ET versus PV ↑apoptotic CECs in PV with >8700 vs.<8700 WBCs |
ADM, adrenomedullin. ANA, anagrelide. APs, activated platelets. ASA, acetylsalycilic acid. CALR, calreticulin. CD, cluster of differentiation. CEC, circulating endothelial cells. cfDNA, cell-free DNA. CK, cytokine(s). CML, chronic myeloid leukemia. CPA, circulating procoagulant activity of plasma. CV risk, cardiovascular risk. ED-1, endothelin-1. ELISA, enzyme-linked immunoassay. EM+IGA, electron microscopy and immunogold analysis. EMPs, endothelial MPs. EMVs, endothelial MVs. ET, essential thrombocythemia. ETP, endogeneous thrombin potential. GMPs, granulocyte-derived MPs. Hb, hemoglobin. Ht, hematocrit. IL-6, interleukin-6. LDH, lactate dehydrogenase. MEVs-CK, monocyte-derived extracellular vesicles. MF, myelofibrosis (unspecified whether primary or secondary). miR, microRNA. MKMVs, megakaryocyte MVs. MMPs, monocyte-derived MPs. MMVs, monocyte MVs. MP, microparticles. MPN, myeloproliferative neoplasms. MPNu, MPN unclassfiable. MVs, microvesicles. PaCS, particulate cytoplasmic structures. PCA, procoagulant activity. PLTs, platelets. PLTMVs, platelet MVs. PMF, primary mielofibrosis. PMAs, platelet microaggregates. PMPs, platelet-derived MPs. PMCs, platelet-monocyte conjugates. PNCs, platelet-neutrophil conjugates. PS, phosphatidylserine. PV, polycythemia vera. RBCs, red blood cells. RMPs, red blood cell MPs. RT, reactive thrombocytosis. RUX, ruxolitinib. SMF, secondary MF. SP, secondary polycythemia. TFMPs, tissue factor-positive MPs. TN, triple-negative. VAF, variant allele frequency. VEFGR2, vascular endothelial growth factor receptor 2. VTE, venous thromboembolism. vs, versus. WB, Western Blot. WBCs, white blood cell count (leukocytes). ↑, increased. ↓, decreased. (+), positive. (−), negative.