Abstract
The molecular mechanisms behind the rupture of intracranial aneurysms remain obscure. MiRNAs are key regulators of a wide array of biological processes altering protein synthesis by binding to target mRNAs. However, variations in miRNA levels in ruptured aneurysmal wall have not been completely examined. We hypothesized that altered miRNA signature in aneurysmal tissues could potentially provide insight into aneurysm pathophysiology. Using a high-throughput miRNA microarray screening approach, we compared the miRNA expression pattern in aneurysm tissues obtained during surgery from patients with aneurysmal subarachnoid hemorrhage (aSAH) with control tissues (GEO accession number GSE161870). We found that the expression of 70 miRNAs was altered. Expressions of the top 10 miRNA were validated, by qRT-PCR and results were correlated with clinical characteristics of aSAH patients. The level of 10 miRNAs (miR-24-3p, miR-26b-5p, miR-27b-3p, miR-125b-5p, miR-143-3p, miR-145-5p, miR-193a-3p, miR-199a-5p, miR-365a-3p/365b-3p, and miR-497-5p) was significantly decreased in patients compared to controls. Expression of miR-125b-5p, miR-143-3p and miR-199a-5p was significantly decreased in patients with poor prognosis and vasospasm. The target genes of few miRNAs were enriched in Transforming growth factor-beta (TGF-β) and Mitogen-activated protein kinases (MAPK) pathways. We found significant negative correlation between the miRNA and mRNA expression (TGF-β1, TGF-β2, SMAD family member 2 (SMAD2), SMAD family member 4 (SMAD4), MAPK1 and MAPK3) in aneurysm tissues. We suggest that miR-26b, miR-199a, miR-497and miR-365, could target multiple genes in TGF-β and MAPK signaling cascades to influence inflammatory processes, extracellular matrix and vascular smooth muscle cell degradation and apoptosis, and ultimately cause vessel wall degradation and rupture.
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Data Availability
The datasets generated during the current study are available from the corresponding author on reasonable request.
Abbreviations
- aSAH:
-
Aneurysmal subarachnoid haemorrhage
- ELISA:
-
Enzyme-linked immunosorbent assay
- GCS:
-
Glasgow coma scale
- GO:
-
Gene ontology
- IA:
-
Intracranial aneurysm
- KEGG:
-
Kyoto encyclopedia of genes and genomes
- MAPK:
-
Mitogen-activated protein kinases
- MiRNA:
-
Micro RNA
- qRT-PCR:
-
Quantitative real time polymerase chain reaction
- SMAD2:
-
SMAD family member 2
- SMAD4:
-
SMAD family member 4
- TGF-β:
-
Transforming growth factor-beta
- WFNS:
-
World federation of neurological surgeons
References
Ambros V (2004) The functions of animal microRNAs. Nature 431(7006):350–355. https://doi.org/10.1038/nature02871
Azuma N, Duzgun SA, Ikeda M, Kito H, Akasaka N, Sasajima T, Sumpio BE (2000) Endothelial cell response to different mechanical forces. J Vasc Surg 32(4):789–794. https://doi.org/10.1067/mva.2000.107989
Bartel DP (2004) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116(2):281–297. https://doi.org/10.1016/s0092-8674(04)00045-5
Bekelis K, Kerley-Hamilton JS, Teegarden A, Tomlinson CR, Kuintzle R, Simmons N, Singer RJ, Roberts DW, Kellis M, Hendrix DA (2016) MicroRNA and gene expression changes in unruptured human cerebral aneurysms. J Neurosurg 125(6):1390–1399. https://doi.org/10.3171/2015.11.JNS151841
Bogatcheva NV, Dudek SM, Garcia JG, Verin AD (2003) Mitogen-activated protein kinases in endothelial pathophysiology. J Investig Med 51(6):341–352. https://doi.org/10.1136/jim-51-06-30
Busch S, Auth E, Scholl F, Huenecke S, Koehl U, Suess B, Steinhilber D (2015) 5-Lipoxygenase is a direct target of miR-19a-3p and miR-125b-5p. J Immunol 194(4):1646–1653. https://doi.org/10.4049/jimmunol.1402163 (Baltimore, Md: 1950)
Cahill PA, Redmond EM (2016) Vascular endothelium—Gatekeeper of vessel health. Atherosclerosis 248:97–109. https://doi.org/10.1016/j.atherosclerosis.2016.03.007
Chalouhi N, Hoh BL, Hasan D (2013) Review of cerebral aneurysm formation, growth, and rupture. Stroke 44(12):3613–3622. https://doi.org/10.1161/strokeaha.113.002390
Cheng Y, Liu X, Yang J, Lin Y, Xu DZ, Lu Q, Deitch EA, Huo Y, Delphin ES, Zhang C (2009) MicroRNA-145, a novel smooth muscle cell phenotypic marker and modulator, controls vascular neointimal lesion formation. Circ Res 105(2):158–166. https://doi.org/10.1161/circresaha.109.197517
Cortez MA, Bueso-Ramos C, Ferdin J, Lopez-Berestein G, Sood AK, Calin GA (2011) MicroRNAs in body fluids–the mix of hormones and biomarkers. Nat Rev Clin Oncol 8(8):467–477. https://doi.org/10.1038/nrclinonc.2011.76
D’Haene B, Mestdagh P, Hellemans J, Vandesompele J (2012) miRNA expression profiling: from reference genes to global mean normalization. Methods Mole Biol (Clifton, NJ) 822:261–272. https://doi.org/10.1007/978-1-61779-427-8_18
D’Souza S (2015) Aneurysmal subarachnoid hemorrhage. J Neurosurg Anesthesiol 27(3):222–240. https://doi.org/10.1097/ana.0000000000000130
Doyle KP, Cekanaviciute E, Mamer LE, Buckwalter MS (2010) TGFbeta signaling in the brain increases with aging and signals to astrocytes and innate immune cells in the weeks after stroke. J Neuroinflammation 7:62. https://doi.org/10.1186/1742-2094-7-62
Dweep H, Gretz N (2015) miRWalk2.0: a comprehensive atlas of microRNA-target interactions. Nature Methods 12(8):697. https://doi.org/10.1038/nmeth.3485
Dweep H, Sticht C, Pandey P, Gretz N (2011) miRWalk–database: prediction of possible miRNA binding sites by “walking” the genes of three genomes. J Biomed Inform 44(5):839–847. https://doi.org/10.1016/j.jbi.2011.05.002
Feigin VL, Rinkel GJ, Lawes CM, Algra A, Bennett DA, van Gijn J, Anderson CS (2005) Risk factors for subarachnoid hemorrhage: an updated systematic review of epidemiological studies. Stroke 36(12):2773–2780. https://doi.org/10.1161/01.STR.0000190838.02954.e8
Fisher CM, Kistler JP, Davis JM (1980) Relation of cerebral vasospasm to subarachnoid hemorrhage visualized by computerized tomographic scanning. Neurosurgery 6(1):1–9. https://doi.org/10.1227/00006123-198001000-00001
Frosen J, Tulamo R, Paetau A, Laaksamo E, Korja M, Laakso A, Niemela M, Hernesniemi J (2012) Saccular intracranial aneurysm: pathology and mechanisms. Acta Neuropathol 123(6):773–786. https://doi.org/10.1007/s00401-011-0939-3
Hong S, Noh H, Chen H, Padia R, Pan ZK, Su SB, Jing Q, Ding HF, Huang S (2013) Signaling by p38 MAPK stimulates nuclear localization of the microprocessor component p68 for processing of selected primary microRNAs. Sci Signal 6(266):ra16. https://doi.org/10.1126/scisignal.2003706
Hu Y, Bock G, Wick G, Xu Q (1998) Activation of PDGF receptor alpha in vascular smooth muscle cells by mechanical stress. FASEB J 12(12):1135–1142. https://doi.org/10.1096/fasebj.12.12.1135
Jamaluddin MS, Weakley SM, Zhang L, Kougias P, Lin PH, Yao Q, Chen C (2011) miRNAs: roles and clinical applications in vascular disease. Expert Rev Mol Diagn 11(1):79–89. https://doi.org/10.1586/erm.10.103
Jennett B, Bond M (1975) Assessment of outcome after severe brain damage. Lancet (Lond, Engl) 1(7905):480–484. https://doi.org/10.1016/s0140-6736(75)92830-5
Jin H, Li C, Ge H, Jiang Y, Li Y (2013) Circulating microRNA: a novel potential biomarker for early diagnosis of intracranial aneurysm rupture a case control study. J Transl Med 11:296. https://doi.org/10.1186/1479-5876-11-296
Karp X, Ambros V (2005) Developmental biology. Encountering microRNAs in cell fate signaling. Science 310(5752):1288–1289. https://doi.org/10.1126/science.1121566 (New York, NY)
Kim S, Izumi Y, Yano M, Hamaguchi A, Miura K, Yamanaka S, Miyazaki H, Iwao H (1998) Angiotensin blockade inhibits activation of mitogen-activated protein kinases in rat balloon-injured artery. Circulation 97(17):1731–1737. https://doi.org/10.1161/01.cir.97.17.1731
Li P, Zhang Q, Wu X, Yang X, Zhang Y, Li Y, Jiang F (2014) Circulating microRNAs serve as novel biological markers for intracranial aneurysms. J Am Heart Assoc 3(5):e000972. https://doi.org/10.1161/jaha.114.000972
Liu D, Han L, Wu X, Yang X, Zhang Q, Jiang F (2014) Genome-wide microRNA changes in human intracranial aneurysms. BMC Neurol 14(1):188. https://doi.org/10.1186/s12883-014-0188-x
Maddahi A, Povlsen GK, Edvinsson L (2012) Regulation of enhanced cerebrovascular expression of proinflammatory mediators in experimental subarachnoid hemorrhage via the mitogen-activated protein kinase kinase/extracellular signal-regulated kinase pathway. J Neuroinflamm 9:274. https://doi.org/10.1186/1742-2094-9-274
Maere S, Heymans K, Kuiper M (2005) BiNGO: a Cytoscape plugin to assess overrepresentation of gene ontology categories in biological networks. Bioinform (Oxf, Engl) 21(16):3448–3449. https://doi.org/10.1093/bioinformatics/bti551
Meeuwsen JAL, van T Hof FNG, van Rheenen W, Rinkel GJE, Veldink JH, Ruigrok YM (2017) Circulating microRNAs in patients with intracranial aneurysms. PLoS ONE 12(5):e0176558. https://doi.org/10.1371/journal.pone.0176558
Morikawa M, Derynck R, Miyazono K (2016) TGF-beta and the TGF-beta family: context-dependent roles in cell and tissue physiology. Cold Spring Harb Perspect in Biol. https://doi.org/10.1101/cshperspect.a021873
Moustakas A, Pardali K, Gaal A, Heldin CH (2002) Mechanisms of TGF-beta signaling in regulation of cell growth and differentiation. Immunol Lett 82(1–2):85–91. https://doi.org/10.1016/s0165-2478(02)00023-8
Paraskevopoulou MD, Georgakilas G, Kostoulas N, Vlachos IS, Vergoulis T, Reczko M, Filippidis C, Dalamagas T, Hatzigeorgiou AG (2013) DIANA-microT web server v5.0: service integration into miRNA functional analysis workflows. Nucleic Acids Res 41(Web Server issue):W169-173. https://doi.org/10.1093/nar/gkt393
Plotnikov A, Zehorai E, Procaccia S, Seger R (2011) The MAPK cascades: signaling components, nuclear roles and mechanisms of nuclear translocation. Biochim Et Biophys Acta 1813(9):1619–1633. https://doi.org/10.1016/j.bbamcr.2010.12.012
Reczko M, Maragkakis M, Alexiou P, Grosse I, Hatzigeorgiou AG (2012) Functional microRNA targets in protein coding sequences. Bioinform (Oxf, Engl) 28(6):771–776. https://doi.org/10.1093/bioinformatics/bts043
Robinson HC, Baker AH (2012) How do microRNAs affect vascular smooth muscle cell biology? Curr Opin Lipidol 23(5):405–411. https://doi.org/10.1097/MOL.0b013e32835719a1
Rosen DS, Macdonald RL (2004) Grading of subarachnoid hemorrhage: modification of the world world federation of neurosurgical societies scale on the basis of data for a large series of patients. Neurosurgery 54(3):566–575 (discussion 575–566)
Santiago-Sim T, Mathew-Joseph S, Pannu H, Milewicz DM, Seidman CE, Seidman JG, Kim DH (2009) Sequencing of TGF-beta pathway genes in familial cases of intracranial aneurysm. Stroke 40(5):1604–1611. https://doi.org/10.1161/strokeaha.108.540245
Santovito D, Mandolini C, Marcantonio P, De Nardis V, Bucci M, Paganelli C, Magnacca F, Ucchino S, Mastroiacovo D, Desideri G, Mezzetti A, Cipollone F (2013) Overexpression of microRNA-145 in atherosclerotic plaques from hypertensive patients. Expert Opin Ther Targets 17(3):217–223. https://doi.org/10.1517/14728222.2013.745512
Steiner T, Juvela S, Unterberg A, Jung C, Forsting M, Rinkel G (2013) European stroke organization guidelines for the management of intracranial aneurysms and subarachnoid haemorrhage. Cerebrovasc Dis (Basel, Switz) 35(2):93–112. https://doi.org/10.1159/000346087
Stylli SS, Adamides AA, Koldej RM, Luwor RB, Ritchie DS, Ziogas J, Kaye AH (2017) miRNA expression profiling of cerebrospinal fluid in patients with aneurysmal subarachnoid hemorrhage. J Neurosurg 126(4):1131. https://doi.org/10.3171/2016.1.jns151454
Supriya M, Christopher R, Indira Devi B, Bhat DI, Shukla D (2020) Circulating MicroRNAs as potential molecular biomarkers for intracranial aneurysmal rupture. Mol Diagn Ther 24(3):351–364. https://doi.org/10.1007/s40291-020-00465-8
Vincze C, Pal G, Wappler EA, Szabo ER, Nagy ZG, Lovas G, Dobolyi A (2010) Distribution of mRNAs encoding transforming growth factors-beta1, -2, and -3 in the intact rat brain and after experimentally induced focal ischemia. J Comp Neurol 518(18):3752–3770. https://doi.org/10.1002/cne.22422
Vlachos IS, Zagganas K, Paraskevopoulou MD, Georgakilas G, Karagkouni D, Vergoulis T, Dalamagas T, Hatzigeorgiou AG (2015) DIANA-miRPath v3.0: deciphering microRNA function with experimental support. Nucleic Acids Res 43(W1):W460-466. https://doi.org/10.1093/nar/gkv403
Yoon S, Seger R (2006) The extracellular signal-regulated kinase: multiple substrates regulate diverse cellular functions. Growth Factors (chur, Switz) 24(1):21–44. https://doi.org/10.1080/02699050500284218
Zacharia BE, Hickman ZL, Grobelny BT, DeRosa P, Kotchetkov I, Ducruet AF, Connolly ES Jr (2010) Epidemiology of aneurysmal subarachnoid hemorrhage. Neurosurg Clin N Am 21(2):221–233. https://doi.org/10.1016/j.nec.2009.10.002
Acknowledgements
The equipment used for this work was provided by the Vision Group on Science and Technology (VGST), Government of Karnataka, India (VGST/CESEM (2014–15)/GRD-311/2015–16). Ms. Supriya M is a recipient of CSIR-SRF fellowship.
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CR, IDB, BDI, SD designed the study. SM and CR collated the data, carried out data analyses and produced the initial draft of the manuscript. KSR provided technical assistance for analysis and interpretation of data. All authors discussed the results, provided critical suggestions, and approved the manuscript.
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Supriya, M., Christopher, R., Devi, B.I. et al. Altered MicroRNA Expression in Intracranial Aneurysmal Tissues: Possible Role in TGF-β Signaling Pathway. Cell Mol Neurobiol 42, 2393–2405 (2022). https://doi.org/10.1007/s10571-021-01121-3
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DOI: https://doi.org/10.1007/s10571-021-01121-3