Plasma microRNAs characterising patients with immune thrombo cytopenic purpura

Bin Zuo; Juping Zhai; Lifang You; Yunxiao Zhao; Jianfeng Yang; Zhen Weng; Lan Dai; Qingyu Wu; Changgeng Ruan; Yang He

doi:10.1160/TH-16-06-0481

Thrombosis and Haemostasis, Table of Contents

Thromb Haemost 2017; 117(07): 1420-1431
DOI: 10.1160/TH-16-06-0481

New Technologies, Diagnostic Tools and Drugs

Schattauer GmbH

Plasma microRNAs characterising patients with immune thrombo cytopenic purpura

Bin Zuo^§

¹MOE Engineering Center of Hematological Disease, Cyrus Tang Hematology Center; MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China

,

Juping Zhai^§

¹MOE Engineering Center of Hematological Disease, Cyrus Tang Hematology Center; MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China

,

Lifang You^§

²Suzhou New & Hi-Tech District Hospital, the Second Affiliated Hospital of Soochow University, Suzhou, China

,

Yunxiao Zhao

¹MOE Engineering Center of Hematological Disease, Cyrus Tang Hematology Center; MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China

,

Jianfeng Yang

¹MOE Engineering Center of Hematological Disease, Cyrus Tang Hematology Center; MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China

,

Zhen Weng

¹MOE Engineering Center of Hematological Disease, Cyrus Tang Hematology Center; MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China

,

Lan Dai

¹MOE Engineering Center of Hematological Disease, Cyrus Tang Hematology Center; MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China

,

Qingyu Wu

¹MOE Engineering Center of Hematological Disease, Cyrus Tang Hematology Center; MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China

,

Changgeng Ruan

¹MOE Engineering Center of Hematological Disease, Cyrus Tang Hematology Center; MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China

,

Yang He

¹MOE Engineering Center of Hematological Disease, Cyrus Tang Hematology Center; MOH Key Lab of Thrombosis and Hemostasis, Jiangsu Institute of Hematology, the First Affiliated Hospital of Soochow University, Suzhou, China

› Author Affiliations

Abstract

Summary

Altered microRNA (miRNA) expression has been reported in patients with immune thrombocytopenic purpura (ITP). However, the detailed expression profiling of cell-free circulating miRNAs in ITP patients has not been fully investigated. In this study, we aimed to examine plasma miRNAs in ITP patients and evaluate their diagnostic values. Plasma samples from 74 ITP patients and 58 healthy controls were obtained and allocated into discovery, validation, and therapy-response sets. Initial screen with a miRNA microarray assay identified 23 miRNAs with different levels between ITP patients and healthy controls (>1.5-fold changes; p<0.01). Subsequent quantitative real-time PCR confirmed eight up-regulated miRNAs (miR-320c, miR-642b-3p, miR-1275, miR-3141, miR-4270, miR-4499, miR-4739 and miR-6126) and three down-regulated miRNAs (miR-144–3p, miR-1281 and miR-3162–3p) in ITP patients. The levels of these circulating miRNAs varied, depending on ITP subtypes, i.e. newly-diagnosed, persistent and chronic ITP, and between treatment responders and non-responders. In receiver operator characteristic analysis, 10 miRNAs had positive diagnostic values (p<0.05) when tested individually. The diagnostic value improved when the miRNAs were analysed as a panel or together with the analysis of anti-platelet autoantibodies. Plasma miR-3162–3p levels were also found to positively correlate with platelet counts in ITP patients (r=0.338, p=0.01). Our results indicate that plasma miRNA profiles are altered in ITP patients and that the differentially expressed miRNAs may be used as biomarkers to improve the diagnosis of ITP.

Keywords

Biomarker - immune thrombocytopenic purpura - microarray - microRNA - platelet autoantibody

Full Text

References

References
1 Cines DB, Bussel JB. How I Treat Idiopathic Thrombocytopenic Purpura (Itp). Blood 2005; 106: 2244-2251.
2 Schwartz RS. Immune Thrombocytopenic Purpura--from Agony to Agonist. N Engl J Med 2007; 357: 2299-2301.
3 Olsson B, Andersson PO, Jernas M. et al. T-Cell-Mediated Cytotoxicity toward Platelets in Chronic Idiopathic Thrombocytopenic Purpura. Nat Med 2003; 9: 1123-1124.
4 Liu B, Zhao H, Poon MC. et al. Abnormality of Cd4(+)Cd25(+) Regulatory T Cells in Idiopathic Thrombocytopenic Purpura. Eur J Haematol 2007; 78: 139-143.
5 Sukati H, Watson HG, Urbaniak SJ. et al. Mapping Helper T-Cell Epitopes on Platelet Membrane Glycoprotein Iiia in Chronic Autoimmune Thrombocytopenic Purpura. Blood 2007; 109: 4528-4538.
6 Ambros V. The Evolution of Our Thinking About Micrornas. Nat Med 2008; 14: 1036-1040.
7 Bartel DP. Micrornas: Genomics, Biogenesis, Mechanism, and Function. Cell 2004; 116: 281-297.
8 Garzon R, Pichiorri F, Palumbo T. et al. Microrna Fingerprints During Human Megakaryocytopoiesis. Proc Natl Acad Sci USA 2006; 103: 5078-5083.
9 Masaki S, Ohtsuka R, Abe Y. et al. Expression Patterns of Micrornas 155 and 451 During Normal Human Erythropoiesis. Biochem Biophys Res Commun 2007; 364: 509-514.
10 Bruchova H, Yoon D, Agarwal AM. et al. Regulated Expression of Micrornas in Normal and Polycythemia Vera Erythropoiesis. Exp Hematol 2007; 35: 1657-1667.
11 Rossi S, Shimizu M, Barbarotto E. et al. Microrna Fingerprinting of Cll Patients with Chromosome 17p Deletion Identify a Mir-21 Score That Stratifies Early Survival. Blood 2010; 116: 945-952.
12 Visone R, Rassenti LZ, Veronese A. et al. Karyotype-Specific Microrna Signature in Chronic Lymphocytic Leukemia. Blood 2009; 114: 3872-3879.
13 Jernas M, Nookaew I, Wadenvik H. et al. Microrna Regulate Immunological Pathways in T-Cells in Immune Thrombocytopenia (Itp). Blood 2013; 121: 2095-2098.
14 McKenzie CG, Guo L, Freedman J. et al. Cellular Immune Dysfunction in Immune Thrombocytopenia (Itp). Br J Haematol 2013; 163: 10-23.
15 Mitchell PS, Parkin RK, Kroh EM. et al. Circulating Micrornas as Stable Blood-Based Markers for Cancer Detection. Proc Natl Acad Sci USA 2008; 105: 10513-10518.
16 Lawrie CH, Gal S, Dunlop HM. et al. Detection of Elevated Levels of Tumour-Associated Micrornas in Serum of Patients with Diffuse Large B-Cell Lymphoma. Br J Haematol 2008; 141: 672-675.
17 Chen X, Ba Y, Ma L. et al. Characterization of Micrornas in Serum: A Novel Class of Biomarkers for Diagnosis of Cancer and Other Diseases. Cell Res 2008; 18: 997-1006.
18 Creemers EE, Tijsen AJ, Pinto YM. Circulating Micrornas: Novel Biomarkers and Extracellular Communicators in Cardiovascular Disease?. Circ Res 2012; 110: 483-495.
19 Bay A, Coskun E, Oztuzcu S. et al. Plasma Microrna Profiling of Pediatric Patients with Immune Thrombocytopenic Purpura. Blood Coagul Fibrinolysis 2014; 25: 379-383.
20 Sui T, Ma L, Li X. et al. Plasma Microrna Profile in Immune Thrombocytopenia: Screening and Verification. Zhonghua Yi Xue Za Zhi 2014; 94: 1083-1086.
21 Rodeghiero F, Stasi R, Gernsheimer T. et al. Standardization of Terminology, Definitions and Outcome Criteria in Immune Thrombocytopenic Purpura of Adults and Children: Report from an International Working Group. Blood 2009; 113: 2386-2393.
22 Tanaka M, Oikawa K, Takanashi M. et al. Down-Regulation of Mir-92 in Human Plasma Is a Novel Marker for Acute Leukemia Patients. PLoS One 2009; 4: e5532.
23 Turchinovich A, Weiz L, Langheinz A. et al. Characterization of Extracellular Circulating Microrna. Nucleic Acids Res 2011; 39: 7223-7233.
24 Kroh EM, Parkin RK, Mitchell PS. et al. Analysis of Circulating Microrna Biomarkers in Plasma and Serum Using Quantitative Reverse Transcription-Pcr (Qrt-Pcr). Methods 2010; 50: 298-301.
25 Zhao Y, Zhu M, Jiang M. et al. An Improved Flow Cytometric Immunobead Array to Detect Autoantibodies in Plasma from Patients with Immune Thrombocytopenic Purpura. Clin Chim Acta 2015; 438: 396-400.
26 Dweep H, Gretz N. Mirwalk2.0: A Comprehensive Atlas of Microrna-Target Interactions. Nat Methods 2015; 12: 697.
27 Chou CH, Chang NW, Shrestha S. et al. Mirtarbase 2016: Updates to the Experimentally Validated Mirna-Target Interactions Database. Nucleic Acids Res 2016; 44 D (01) D239-D247.
28 Porcelijn L, Huiskes E, Oldert G. et al. Detection of Platelet Autoantibodies Revisited to Identify Immune Thrombocytopenia. Blood 2015; 126: 1142-1142.
29 He Y, Zhao YX, Zhu MQ. et al. Detection of Autoantibodies against Platelet Glycoproteins in Patients with Immune Thrombocytopenic Purpura by Flow Cytometric Immunobead Array. Clin Chim Acta 2013; 415: 176-180.
30 Vlachos IS, Zagganas K, Paraskevopoulou MD. et al. Diana-Mirpath V3.0: Deciphering Microrna Function with Experimental Support. Nucleic Acids Res 2015; 43 W (01) W460-W466.
31 George JN, Woolf SH, Raskob GE. et al. Idiopathic Thrombocytopenic Purpura: A Practice Guideline Developed by Explicit Methods for the American Society of Hematology. Blood 1996; 88: 3-40.
32 Provan D, Stasi R, Newland AC. et al. International Consensus Report on the Investigation and Management of Primary Immune Thrombocytopenia. Blood 2010; 115: 168-186.
33 Etheridge A, Lee I, Hood L. et al. Extracellular Microrna: A New Source of Biomarkers. Mutat Res 2011; 717: 85-90.
34 van Rooij E, Marshall WS, Olson EN. Toward Microrna-Based Therapeutics for Heart Disease: The Sense in Antisense. Circ Res 2008; 103: 919-928.
35 Martin IW, Steinmetz HB, Lefferts CL. et al. Evaluation of the Cobas 4800 Hpv Test for Detecting High-Risk Human Papilloma-Virus in Cervical Cytology Specimens. Pathogens 2012; 1: 30-36.
36 Smith LK, Shah RR, Cidlowski JA. Glucocorticoids Modulate Microrna Expression and Processing During Lymphocyte Apoptosis. J Biol Chem 2010; 285: 36698-36708.
37 Davis TE, Kis-Toth K, Szanto A. et al. Glucocorticoids Suppress T Cell Function by up-Regulating Microrna-98. Arthritis Rheum 2013; 65: 1882-1890.
38 Cook NR. Statistical Evaluation of Prognostic Versus Diagnostic Models: Beyond the Roc Curve. Clin Chem 2008; 54: 17-23.
39 Perkins NJ, Schisterman EF. The Inconsistency of “Optimal„ Cutpoints Obtained Using Two Criteria Based on the Receiver Operating Characteristic Curve. Am J Epidemiol 2006; 163: 670-675.
40 Wang Y, Yang L, Li P. et al. Circulating Microrna Signatures Associated with Childhood Asthma. Clin Lab 2015; 61: 467-474.
41 Fang C, Lu W, Li C. et al. Mir-3162-3p Is a Novel Microrna That Exacerbates Asthma by Regulating Beta-Catenin. PLoS One 2016; 11: e0149257.
42 Williams AE, Choi K, Chan AL. et al. Sjogren’s Syndrome-Associated Micrornas in Cd14(+) Monocytes Unveils Targeted Tgfbeta Signaling. Arthritis Res Ther 2016; 18: 95.
43 Glass CK, Saijo K. Nuclear Receptor Transrepression Pathways That Regulate Inflammation in Macrophages and T Cells. Nat Rev Immunol 2010; 10: 365-376.

Supplementary Material

Online Supplementary Material (PDF)