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DOI: 10.1160/TH16-08-0606
Distinctive expression signatures of serum microRNAs in ischaemic stroke and transient ischaemic attack patients
Financial support: This work was supported by the National Natural Science Foundation of China (81271904, 81401742, 81572073 and 81572074), the Special-funded Program on National Key Scientific Instruments and Equipment Development of China (2012YQ03026109), the Medical Scientific Research Foundation of Nanjing Military Command (12Z28) and the Jinling Hospital Foundation (2014051).Publication History
Received:
05 August 2016
Accepted after major revision:
09 February 2017
Publication Date:
28 November 2017 (online)
Summary
Circulating microRNAs (miRNAs) have recently emerged as promising biomarkers for ischaemic stroke (IS). However, the expression patterns of specific miRNAs in transient ischaemic attack (TIA) patients have not been investigated. Their predictive values for the presence of IS and TIA and their relationships to the neurological deficit severity of IS and the subsequent stroke risk after TIA remain unclear exactly. In this study, 754 miRNAs were initially screened by the TaqMan Low Density Array (TLDA) in two pooled serum samples from 50 IS patients and 50 controls. Markedly altered miRNAs were subsequently validated by individual quantitative reverse-transcription PCR (qRT-PCR) assays first in the same cohort of TLDA and further confirmed in another larger cohort including 177 IS, 81 TIA patients and 42 controls. Consequently, TLDA screening showed that 71 miRNAs were up-regulated and 49 miRNAs were down-regulated in IS patients. QRT-PCR validation confirmed that serum levels of miR-23b-3p, miR-29b-3p, miR-181a-5p and miR-21–5p were significantly increased in IS patients. Strikingly, serum levels of miR-23b-3p, miR-29b-3p and miR-181a-5p were also significantly elevated in TIA patients. Furthermore, up-regulated miR-23b-3p, miR-29b-3p and miR-21–5p could clearly differentiate between IS and TIA patients. Logistic regression and receiver-operating characteristic curve analyses demonstrated that these altered miRNAs may function as predictive and discriminative biomarkers for IS and TIA, and their distinctive expression signatures may contribute to assessing neurological deficit severity of IS and subsequent stroke risk after TIA.
Supplementary Material to this article is available online at www.thrombosis-online.com.
# These authors contributed equally to this work.
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References
- 1 Mozaffarian D, Benjamin EJ, Go AS. et al. Heart disease and stroke statistics--2015 update: a report from the American Heart Association. Circulation 2015; 131: e29-322.
- 2 Liu L, Wang D, Wong KS. et al. Stroke and stroke care in China: huge burden, significant workload, and a national priority. Stroke 2011; 42: 3651-3654.
- 3 Saenger AK, Christenson RH. Stroke biomarkers: progress and challenges for diagnosis, prognosis, differentiation, and treatment. Clin Chem 2010; 56: 21-33.
- 4 Easton JD, Saver JL, Albers GW. et al. Definition and evaluation of transient ischaemic attack: a scientific statement for healthcare professionals from the American Heart Association/American Stroke Association Stroke Council; Council on Cardiovascular Surgery and Anesthesia; Council on Cardiovascular Radiology and Intervention; Council on Cardiovascular Nursing; and the Interdisciplinary Council on Peripheral Vascular Disease. The American Academy of Neurology affirms the value of this statement as an educational tool for neurologists. Stroke 2009; 40: 2276-2293
- 5 Mullins ME, Schaefer PW, Sorensen AG. et al. CT and conventional and diffusion-weighted MR imaging in acute stroke: study in 691 patients at presentation to the emergency department. Radiology 2002; 224: 353-360.
- 6 Ay H, Oliveira-Filho J, Buonanno FS. et al. ’Footprints’ of transient ischaemic attacks: a diffusion-weighted MRI study. Cerebrovasc Dis 2002; 14: 177-186.
- 7 Whiteley W, Wardlaw J, Dennis M. et al. The use of blood biomarkers to predict poor outcome after acute transient ischaemic attack or ischaemic stroke. Stroke 2012; 43: 86-91.
- 8 Prugger C, Luc G, Haas B. et al. Multiple biomarkers for the prediction of ischaemic stroke: the PRIME study. Arterioscler Thromb Vasc Biol 2013; 33: 659-666.
- 9 van Rooij E. The art of microRNA research. Circ Res 2011; 108: 219-234.
- 10 Mendell JT, Olson EN. MicroRNAs in stress signaling and human disease. Cell 2012; 148: 1172-1187.
- 11 Creemers EE, Tijsen AJ, Pinto YM. Circulating microRNAs: novel biomarkers and extracellular communicators in cardiovascular disease?. Circ Res 2012; 110: 483-495.
- 12 Schwarzenbach H, Nishida N, Calin GA. et al. Clinical relevance of circulating cell-free microRNAs in cancer. Nat Rev Clin Oncol 2014; 11: 145-156.
- 13 Orenes-Piñero E, Montoro-García S, Patel JV. et al. Role of microRNAs in cardiac remodelling: new insights and future perspectives. Int J Cardiol 2013; 167: 1651-1659.
- 14 Im HI, Kenny PJ. MicroRNAs in neuronal function and dysfunction. Trends Neurosci 2012; 35: 325-334.
- 15 Bhalala OG, Srikanth M, Kessler JA. The emerging roles of microRNAs in CNS injuries. Nat Rev Neurol 2013; 09: 328-339.
- 16 Doeppner TR, Doehring M, Bretschneider E. et al. MicroRNA-124 protects against focal cerebral ischaemia via mechanisms involving Usp14-dependent REST degradation. Acta Neuropathol 2013; 126: 251-265.
- 17 Irmady K, Jackman KA, Padow VA. et al. Mir-592 regulates the induction and cell death-promoting activity of p75NTR in neuronal ischaemic injury. J Neurosci 2014; 34: 3419-3428.
- 18 Qu Y, Wu J, Chen D. et al. MiR-139-5p inhibits HGTD-P and regulates neuronal apoptosis induced by hypoxia-ischaemia in neonatal rats. Neurobiol Dis 2014; 63: 184-193.
- 19 Tan KS, Armugam A, Sepramaniam S. et al. Expression profile of MicroRNAs in young stroke patients. PLoS One 2009; 04: e7689.
- 20 Sepramaniam S, Tan JR, Tan KS. et al. Circulating microRNAs as biomarkers of acute stroke. Int J Mol Sci 2014; 15: 1418-1432.
- 21 Wang W, Sun G, Zhang L. et al. Circulating microRNAs as novel potential biomarkers for early diagnosis of acute stroke in humans. J Stroke Cerebrovasc Dis 2014; 23: 2607-2613.
- 22 WHO Task Force on Stroke and other Cerebrovascular Disorders. Stroke--1989: Recommendations on stroke prevention, diagnosis, and therapy. Stroke 1989; 20: 1407-1421.
- 23 Sarker SJ, Rudd AG, Douiri A. et al. Comparison of 2 extended activities of daily living scales with the Barthel Index and predictors of their outcomes: cohort study within the South London Stroke Register (SLSR). Stroke 2012; 43: 1362-1369.
- 24 Kiyohara T, Kamouchi M, Kumai Y. et al. ABCD3 and ABCD3-I scores are superior to ABCD2 score in the prediction of short- and long-term risks of stroke after transient ischaemic attack. Stroke 2014; 45: 418-425.
- 25 Luo Y, Wang C, Chen X. et al. Increased serum and urinary micrornas in children with idiopathic nephrotic syndrome. Clin Chem 2013; 59: 658-666.
- 26 Wu J, Song J, Wang C. et al. Identification of serum microRNAs for cardiovascular risk stratification in dyslipidemia subjects. Int J Cardiol 2014; 172: 232-234.
- 27 Wang C, Hu J, Lu M. et al. A panel of five serum miRNAs as a potential diagnostic tool for early-stage renal cell carcinoma. Sci Rep 2015; 05: 7610.
- 28 Chen X, Liang H, Guan D. et al. A combination of Let-7d, Let-7g and Let-7i serves as a stable reference for normalisation of serum microRNAs. PLoS One 2013; 08: e79652.
- 29 Chio CC, Lin JW, Cheng HA. et al. MicroRNA-210 targets antiapoptotic Bcl-2 expression and mediates hypoxia-induced apoptosis of neuroblastoma cells. Arch Toxicol 2013; 87: 459-468.
- 30 Li D, Qu Y, Mao M. et al. Involvement of the PTEN-AKT-FOXO3a pathway in neuronal apoptosis in developing rat brain after hypoxia-ischaemia. J Cereb Blood Flow Metab 2009; 29: 1903-1913.
- 31 Carloni S, Albertini MC, Galluzzi L. et al. Melatonin reduces endoplasmic reticulum stress and preserves sirtuin 1 expression in neuronal cells of newborn rats after hypoxia-ischaemia. J Pineal Res 2014; 57: 192-199.
- 32 Siegel C, Li J, Liu F. et al. miR-23a regulation of X-linked inhibitor of apoptosis (XIAP) contributes to sex differences in the response to cerebral ischaemia. Proc Natl Acad Sci USA 2011; 108: 11662-11667.
- 33 Pikula A, Beiser AS, Chen TC. et al. Serum brain-derived neurotrophic factor and vascular endothelial growth factor levels are associated with risk of stroke and vascular brain injury: Framingham Study. Stroke 2013; 44: 2768-2775.
- 34 Gelderblom M, Daehn T, Schattling B. et al. Plasma levels of neuron specific enolase quantify the extent of neuronal injury in murine models of ischaemic stroke and multiple sclerosis. Neurobiol Dis 2013; 59: 177-182.
- 35 Tsai PC, Liao YC, Wang YS. et al. Serum microRNA-21 and microRNA-221 as potential biomarkers for cerebrovascular disease. J Vasc Res 2013; 50: 346-354.
- 36 Zhang L, Dong LY, Li YJ. et al. miR-21 represses FasL in microglia and protects against microglia-mediated neuronal cell death following hypoxia/ischaemia. Glia 2012; 60: 1888-1895.
- 37 Buller B, Liu X, Wang X. et al. MicroRNA-21 protects neurons from ischaemic death. FEBS J 2010; 277: 4299-4307.
- 38 Chen Q, Xu Ji, Li L. et al. MicroRNA-23a/b and microRNA-27a/b suppress Apaf-1 protein and alleviate hypoxia-induced neuronal apoptosis. Cell Death Dis 2014; 05: e1132.
- 39 Moon JM, Xu L, Giffard RG. Inhibition of microRNA-181 reduces forebrain ischaemia-induced neuronal loss. J Cereb Blood Flow Metab 2013; 33: 1976-1982.
- 40 Ouyang YB, Lu Y, Yue S. et al. miR-181 regulates GRP78 and influences outcome from cerebral ischaemia in vitro and in vivo. Neurobiol Dis 2012; 45: 555-563.
- 41 Shi G, Liu Y, Liu T. et al. Upregulated miR-29b promotes neuronal cell death by inhibiting Bcl2L2 after ischaemic brain injury. Exp Brain Res 2012; 216: 225-230.
- 42 Khanna S, Rink C, Ghoorkhanian R. et al. Loss of miR-29b following acute ischaemic stroke contributes to neural cell death and infarct size. J Cereb Blood Flow Metab 2013; 33: 1197-1206.
- 43 Chen X, Liang H, Zhang J. et al. Secreted microRNAs: a new form of intercellular communication. Trends Cell Biol 2012; 22: 125-132.
- 44 Turchinovich A, Weiz L, Burwinkel B. Extracellular miRNAs: the mystery of their origin and function. Trends Biochem Sci 2012; 37: 460-465.
- 45 Kwah LK, Harvey LA, Diong J. et al. Models containing age and NIHSS predict recovery of ambulation and upper limb function six months after stroke: an observational study. J Physiother 2013; 59: 189-197.
- 46 Zhao H, Wang J, Gao L. et al. MiRNA-424 protects against permanent focal cerebral ischaemia injury in mice involving suppressing microglia activation. Stroke 2013; 44: 1706-1713.
- 47 Liu DZ, Jickling GC, Ander BP. et al. Elevating microRNA-122 in blood improves outcomes after temporary middle cerebral artery occlusion in rats. J Cereb Blood Flow Metab 2016; 36: 1374-1383.