Diagnostic potential of plasma microRNA signatures in patients with deep-vein thrombosis
Xiao Wang
1
Center for Primary Health Care Research, Lund University/Region Skåne, Sweden
,
Kristina Sundquist
1
Center for Primary Health Care Research, Lund University/Region Skåne, Sweden
,
Johan L. Elf
2
Vascular Centers, Lund University, Malmö, University Hospital Malmö, Sweden
,
Karin Strandberg
3
Department of Clinical Chemistry, Lund University, Malmö, University Hospital, Malmö, Sweden
,
Peter J. Svensson
4
Department of Coagulation Disorders, Lund University, Malmö, University Hospital, Malmö, Sweden
,
Anna Hedelius
1
Center for Primary Health Care Research, Lund University/Region Skåne, Sweden
,
Karolina Palmér
1
Center for Primary Health Care Research, Lund University/Region Skåne, Sweden
,
Ashfaque A. Memon
1
Center for Primary Health Care Research, Lund University/Region Skåne, Sweden
,
Jan Sundquist
1
Center for Primary Health Care Research, Lund University/Region Skåne, Sweden
,
Bengt Zöller
1
Center for Primary Health Care Research, Lund University/Region Skåne, Sweden
› Author AffiliationsFinancial support: This work was supported by grants awarded to Bengt Zöller by the Swedish Heart-Lung Foundation, ALF funding from Region Skåne awarded to Bengt Zöller, Jan Sundquist, and Kristina Sundquist, and grants awarded to Bengt Zöller, Kristina Sundquist and Jan Sundquist by the Swedish Research Council.
For excluding deep-vein thrombosis (DVT), a negative D-dimer and low clinical probability are used to rule out DVT. Circulating microRNAs (miRNAs) are stably present in the plasma, serum and other body fluids. Their diagnostic function has been investigated in many diseases but not in DVT. The aims of present study were to assess the diagnostic ability of plasma miRNAs in DVT and to examine their correlation with known markers of hypercoagulability, such as D-dimer and APC-PCI complex. Plasma samples were obtained from 238 patients (aged 16–95 years) with suspected DVT included in a prospective multicentre management study (SCORE). We first performed miRNA screening of plasma samples from three plasma pools containing plasma from 12 patients with DVT and three plasma pools containing plasma from 12 patients without DVT using a microRNA Ready-to-use PCR Panel comprising 742 miRNA primer sets. Thirteen miRNAs that differentially expressed were further investigated by quantitative real-time (qRT)-PCR in the entire cohort. The plasma level of miR-424–5p (p=0.01) were significantly higher, whereas the levels of miR-136–5p (p=0.03) were significantly lower in DVT patients compared to patients without DVT. Receiver-operating characteristic curve analysis showed the area under the curve (AUC) values of 0.63 for miR-424–5p and 0.60 for miR-136–5p. The plasma level of miR-424–5p was associated with both D-dimer and APC-PCI complex levels (p<0.0001 and p=0.001, respectively). In conclusions, these findings indicate that certain miRNAs are associated with DVT and markers of hypercoagulability, though their diagnostic abilities are probably too low.
Supplementary Material to this article is available online at www.thrombosis-online.com.
1
Silverstein MD,
Heit JA,
Mohr DN.
et al. Trends in the incidence of deep vein thrombosis and pulmonary embolism: a 25-year population-based study. Arch Intern Med 1998; 158: 585-593.
2
Naess IA,
Christiansen SC,
Romundstad P.
et al. Incidence and mortality of venous thrombosis: a population-based study. J Thromb Haemost 2007; 5: 692-699.
6
Elf JL,
Strandberg K,
Nilsson C.
et al. Clinical probability assessment and D-dimer determination in patients with suspected deep vein thrombosis, a prospective multicenter management study. Thromb Res 2009; 123: 612-616.
7
Elf JL,
Strandberg K,
Svensson PJ..
The diagnostic performance of APC-PCI complex determination compared to D-dimer in the diagnosis of deep vein thrombosis. J Thromb Thrombol 2010; 29: 465-470.
8
Lippi G,
Cervellin G,
Franchini M.
et al. Biochemical markers for the diagnosis of venous thromboembolism: the past, present and future. J Thromb Thrombol 2010; 30: 459-471.
10
Miranda KC,
Huynh T,
Tay Y.
et al. A pattern-based method for the identification of MicroRNA binding sites and their corresponding heteroduplexes. Cell 2006; 126: 1203-1217.
12
Arroyo JD,
Chevillet JR,
Kroh EM.
et al. Argonaute2 complexes carry a population of circulating microRNAs independent of vesicles in human plasma. Proc Natl Acad Sci USA 2011; 108: 5003-5008.
13
Vickers KC,
Palmisano BT,
Shoucri BM.
et al. MicroRNAs are transported in plasma and delivered to recipient cells by high-density lipoproteins. Nature Cell Biol 2011; 13: 423-433.
14
Zampetaki A,
Kiechl S,
Drozdov I.
et al. Plasma microRNA profiling reveals loss of endothelial miR-126 and other microRNAs in type 2 diabetes. Circ Res 2010; 107: 810-817.
17
Qin J,
Liang H,
Shi D.
et al. A panel of microRNAs as a new biomarkers for the detection of deep vein thrombosis. J Thromb Thrombol 2015; 39: 215-221.
18
Xiao J,
Jing ZC,
Ellinor PT.
et al. MicroRNA-134 as a potential plasma biomarker for the diagnosis of acute pulmonary embolism. J Transl Med 2011; 9: 159.
19
Starikova I,
Jamaly S,
Sorrentino A.
et al. Differential expression of plasma miRNAs in patients with unprovoked venous thromboembolism and healthy control individuals. Thromb Res 2015; 136: 566-572.
20
Wells PS,
Anderson DR,
Bormanis J.
et al. Value of assessment of pretest probability of deep-vein thrombosis in clinical management. Lancet 1997; 350: 1795-1798.
22
Strandberg K,
Kjellberg M,
Knebel R.
et al. A sensitive immunochemical assay for measuring the concentration of the activated protein C-protein C inhibitor complex in plasma: use of a catcher antibody specific for the complexed/cleaved form of the inhibitor. Thromb Haemost 2001; 86: 604-610.
23
Wang X,
Sundquist J,
Zoller B.
et al. Determination of 14 circulating microRNAs in Swedes and Iraqis with and without diabetes mellitus type 2. PloS one 2014; 9: e86792.
24
Kendziorski C,
Irizarry RA,
Chen KS.
et al. On the utility of pooling biological samples in microarray experiments. Proc Natl Acad Sci USA 2005; 102: 4252-4257.
25
Kok MG,
Halliani A,
Moerland PD.
et al. Normalisation panels for the reliable quantification of circulating microRNAs by RT-qPCR. FASEB J 2015; 29: 3853-3862.
29
Blondal T,
Jensby Nielsen S,
Baker A.
et al. Assessing sample and miRNA profile quality in serum and plasma or other biofluids. Methods 2013; 59: S1-6.
30
Song J,
Bai Z,
Han W.
et al. Identification of suitable reference genes for qPCR analysis of serum microRNA in gastric cancer patients. Digest Dis Sci 2012; 57: 897-904.
31
Shen Y,
Li Y,
Ye F.
et al. Identification of miR-23a as a novel microRNA normalizer for relative quantification in human uterine cervical tissues. Exp Mol Med 2011; 43: 358-366.
32
Rodosthenous RS,
Coull BA,
Lu Q.
et al. Ambient particulate matter and microRNAs in extracellular vesicles: a pilot study of older individuals. Particle Fibre Toxicol 2016; 13: 13.
33
Polytarchou C,
Oikonomopoulos A,
Mahurkar S.
et al. Assessment of Circulating MicroRNAs for the Diagnosis and Disease Activity Evaluation in Patients with Ulcerative Colitis by Using the Nanostring Technology. Inflamm Bowel Dis 2015; 21: 2533-2539.
34
Satoh J,
Kino Y,
Niida S..
MicroRNA- Seq Data Analysis Pipeline to Identify Blood Biomarkers for Alzheimer's Disease from Public Data. Biomarker Insights 2015; 10: 21-31.
35
Park B,
Messina L,
Dargon P.
et al. Recent trends in clinical outcomes and resource utilisation for pulmonary embolism in the United States: findings from the nationwide inpatient sample. Chest 2009; 136: 983-990.
36
Koster T,
Blann AD,
Briet E.
et al. Role of clotting factor VIII in effect of von Wille-brand factor on occurrence of deep-vein thrombosis. Lancet 1995; 345: 152-155.
37
Wells PS,
Anderson DR,
Rodger M.
et al. Evaluation of D-dimer in the diagnosis of suspected deep-vein thrombosis. N Engl J Med 2003; 349: 1227-1235.
40
Lu TP,
Lee CY,
Tsai MH.
et al. miRSystem: an integrated system for characterizing enriched functions and pathways of microRNA targets. PloS one 2012; 7: e42390.
41
Wang X..
Improving microRNA target prediction by modeling with unambiguously identified microRNA-target pairs from CLIP- Ligation studies. Bioinformatics. 2016 Epub ahead of print.
43
Forrest AR,
Kanamori-Katayama M,
Tomaru Y.
et al. Induction of microRNAs, mir-155, mir-222, mir-424 and mir-503, promotes monocytic differentiation through combinatorial regulation. Leukemia 2010; 24: 460-466.
44
Sarkar S,
Dey BK,
Dutta A..
MiR-322/424 and –503 are induced during muscle differentiation and promote cell cycle quiescence and differentiation by down-regulation of Cdc25A. Mol Biol Cell 2010; 21: 2138-2149.
45
Rosa A,
Ballarino M,
Sorrentino A.
et al. The interplay between the master transcription factor PU.1 and miR-424 regulates human monocyte/macrophage differentiation. Proc Natl Acad Sci USA 2007; 104: 19849-19854.
47
Kim J,
Kang Y,
Kojima Y.
et al. An endothelial apelin-FGF link mediated by miR-424 and miR-503 is disrupted in pulmonary arterial hypertension. Nature Med 2013; 19: 74-82.
49
Lau P,
Bossers K,
Janky R.
et al. Alteration of the microRNA network during the progression of Alzheimer's disease. EMBO Mol Med 2013; 5: 1613-1634.
