Abstract
Circular RNAs (circRNAs) are highly enriched in the brain and involved in many types of central nervous system pathologies. Herein, this study aimed to investigate the role and mechanism of circ_0007290 in ischemic stroke. The oxygen-glucose deprivation (OGD) model was established with the HCN-2 cells in vitro. Levels of genes and proteins was detected by quantitative real-time polymerase chain reaction (qRT-PCR) and Western blot. In vitro experiments were conducted using cell counting kit-8 (CCK-8) assay, EdU (5-ethynyl-2’-deoxyuridine) assay, flow cytometry and ELISA, respectively. The levels of lactate dehydrogenase (LDH) were measured using the commercial kit. RNA pull-down and dual-luciferase reporter assay were used to identify the target relationship between miR-496 and circ_0007290 or PDCD4 (programmed cell death protein 4). Circ_0007290 expression was elevated in acute ischemic stroke (AIS) patients and OGD-induced cell injury model. OGD stimulation induced neuronal apoptosis, promoted LDH release, and enhanced inflammation in HCN-2 cells, which all were reversed by the knockdown of circ_0007290. Mechanistically, circ_0007290 served as a sponge for miR-496 to relieve the repression of miR-496 on the expression of its target PDCD4. Moreover, miR-496 inhibition or PDCD4 overexpression abolished the inhibitory effects of circ_0007290 knockdown OGD-evoked neuronal injury. Knockdown of circ_0007290 alleviated OGD-induced neuronal injury by regulating miR-496/PDCD4 axis, providing a novel insight into the pathology of ischemic stroke.
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Data availability
The analyzed data sets generated during the present study are available from the corresponding author on reasonable request.
References
Ashwal-Fluss R, Meyer M, Pamudurti NR, Ivanov A, Bartok O, Hanan M, Evantal N, Memczak S, Rajewsky N, Kadener S (2014) circRNA biogenesis competes with pre-mRNA splicing. Mol Cell 56:55–66. https://doi.org/10.1016/j.molcel.2014.08.019
Bian L, Zhi X, Ma L, Zhang J, Chen P, Sun S, Li J, Sun Y, Qin J (2018) Hsa_circRNA_103809 regulated the cell proliferation and migration in colorectal cancer via miR-532-3p / FOXO4 axis. Biochem Biophys Res Commun 505:346–352. https://doi.org/10.1016/j.bbrc.2018.09.073
Chen J, Cui C, Yang X, Xu J, Venkat P, Zacharek A, Yu P, Chopp M (2017) MiR-126 affects brain-heart interaction after cerebral ischemic stroke. Transl Stroke Res 8:374–385. https://doi.org/10.1007/s12975-017-0520-z
Donnan GA, Fisher M, Macleod M, Davis SM (2008) Stroke Lancet 371:1612–1623. https://doi.org/10.1016/s0140-6736(08)60694-7
Hacke W, Kaste M, Bluhmki E, Brozman M, Dávalos A, Guidetti D, Larrue V, Lees KR, Medeghri Z, Machnig T, Schneider D, von Kummer R, Wahlgren N, Toni D (2008) Thrombolysis with alteplase 3 to 4.5 hours after acute ischemic stroke. N Engl J Med 359:1317–1329. https://doi.org/10.1056/NEJMoa0804656
Han B, Zhang Y, Zhang Y, Bai Y, Chen X, Huang R, Wu F, Leng S, Chao J, Zhang JH, Hu G, Yao H (2018) Novel insight into circular RNA HECTD1 in astrocyte activation via autophagy by targeting MIR142-TIPARP: implications for cerebral ischemic stroke. Autophagy 14:1164–1184. https://doi.org/10.1080/15548627.2018.1458173
Hanan M, Soreq H, Kadener S (2017) CircRNAs in the brain. RNA Biol 14:1028–1034. https://doi.org/10.1080/15476286.2016.1255398
Hansen TB, Jensen TI, Clausen BH, Bramsen JB, Finsen B, Damgaard CK, Kjems J (2013) Natural RNA circles function as efficient microRNA sponges. Nature 495:384–388. https://doi.org/10.1038/nature11993
Hou LD, Zhang J (2017) Circular RNAs: An emerging type of RNA in cancer. Int J Immunopathol Pharmacol 30:1–6. https://doi.org/10.1177/0394632016686985
Jin Y, Ni S (2020) miR-496 remedies hypoxia reoxygenation-induced H9c2 cardiomyocyte apoptosis via Hook3-targeted PI3k/Akt/mTOR signaling pathway activation. J Cell Biochem 121:698–712. https://doi.org/10.1002/jcb.29316
Jobe EM, McQuate AL, Zhao X (2012) Crosstalk among epigenetic pathways regulates neurogenesis. Front Neurosci 6:59. https://doi.org/10.3389/fnins.2012.00059
Mahmoudi E, Cairns MJ (2019) Circular RNAs are temporospatially regulated throughout development and ageing in the rat. Sci Rep 9:2564. https://doi.org/10.1038/s41598-019-38860-9
Mehta SL, Dempsey RJ, Vemuganti R (2020) Role of circular RNAs in brain development and CNS diseases. Prog Neurobiol 186:101746. https://doi.org/10.1016/j.pneurobio.2020.101746
Memczak S, Jens M, Elefsinioti A, Torti F, Krueger J, Rybak A, Maier L, Mackowiak SD, Gregersen LH, Munschauer M, Loewer A, Ziebold U, Landthaler M, Kocks C, le Noble F, Rajewsky N (2013) Circular RNAs are a large class of animal RNAs with regulatory potency. Nature 495:333–338. https://doi.org/10.1038/nature11928
Nieto-Diaz M, Esteban FJ, Reigada D, Muñoz-Galdeano T, Yunta M, Caballero-López M, Navarro-Ruiz R, Del Águila A, Maza RM (2014) MicroRNA dysregulation in spinal cord injury: causes, consequences and therapeutics. Front Cell Neurosci 8:53. https://doi.org/10.3389/fncel.2014.00053
Patop IL, Kadener S (2018) circRNAs in Cancer. Curr Opin Genet Dev 48:121–127. https://doi.org/10.1016/j.gde.2017.11.007
Rago L, Beattie R, Taylor V, Winter J (2014) miR379-410 cluster miRNAs regulate neurogenesis and neuronal migration by fine-tuning N-cadherin. Embo j 33:906–920. https://doi.org/10.1002/embj.201386591
Ribeiro PW, Cola PC, Gatto AR, da Silva RG, Luvizutto GJ, Braga GP, Schelp AO, de Arruda Henry MA, Bazan R (2015) Relationship between Dysphagia, National Institutes of Health Stroke Scale Score, and Predictors of Pneumonia after Ischemic Stroke. J Stroke Cerebrovasc Dis 24:2088–2094. https://doi.org/10.1016/j.jstrokecerebrovasdis.2015.05.009
Rybak-Wolf A, Stottmeister C, Glažar P, Jens M, Pino N, Giusti S, Hanan M, Behm M, Bartok O, Ashwal-Fluss R, Herzog M, Schreyer L, Papavasileiou P, Ivanov A, Öhman M, Refojo D, Kadener S, Rajewsky N (2015) Circular RNAs in the mammalian brain are highly abundant, conserved, and dynamically expressed. Mol Cell 58:870–885. https://doi.org/10.1016/j.molcel.2015.03.027
Salzman J, Gawad C, Wang PL, Lacayo N, Brown PO (2012) Circular RNAs are the predominant transcript isoform from hundreds of human genes in diverse cell types. PLoS ONE 7:e30733. https://doi.org/10.1371/journal.pone.0030733
Sheth KN, Smith EE, Grau-Sepulveda MV, Kleindorfer D, Fonarow GC, Schwamm LH (2015) Drip and ship thrombolytic therapy for acute ischemic stroke: use, temporal trends, and outcomes. Stroke 46:732–739. https://doi.org/10.1161/strokeaha.114.007506
Tobin MK, Bonds JA, Minshall RD, Pelligrino DA, Testai FD, Lazarov O (2014) Neurogenesis and inflammation after ischemic stroke: what is known and where we go from here. J Cereb Blood Flow Metab 34:1573–1584. https://doi.org/10.1038/jcbfm.2014.130
Vijayan M, Reddy PH (2016a) Peripheral biomarkers of stroke: Focus on circulatory microRNAs. Biochim Biophys Acta 1862:1984-1993. https://doi.org/10.1016/j.bbadis.2016.08.003
Vijayan M, Reddy PH (2016) Stroke, vascular dementia, and Alzheimer’s Disease: molecular links. J Alzheimers Dis 54:427–443. https://doi.org/10.3233/jad-160527
Vijayan M, Kumar S, Yin X, Zafer D, Chanana V, Cengiz P, Reddy PH (2018) Identification of novel circulatory microRNA signatures linked to patients with ischemic stroke. Hum Mol Genet 27:2318–2329. https://doi.org/10.1093/hmg/ddy136
Winter J (2015) MicroRNAs of the miR379-410 cluster: New players in embryonic neurogenesis and regulators of neuronal function. Neurogenesis (Austin) 2:e1004970. https://doi.org/10.1080/23262133.2015.1004970
Wu F, Han B, Wu S, Yang L, Leng S, Li M, Liao J, Wang G, Ye Q, Zhang Y, Chen H, Chen X, Zhong M, Xu Y, Liu Q, Zhang JH, Yao H (2019) Circular RNA TLK1 aggravates neuronal injury and neurological deficits after ischemic stroke via miR-335-3p/TIPARP. J Neurosci 39:7369–7393. https://doi.org/10.1523/jneurosci.0299-19.2019
Yan H, Rao J, Yuan J, Gao L, Huang W, Zhao L, Ren J (2017) Long non-coding RNA MEG3 functions as a competing endogenous RNA to regulate ischemic neuronal death by targeting miR-21/PDCD4 signaling pathway. Cell Death Dis 8:3211. https://doi.org/10.1038/s41419-017-0047-y
Yang HS, Jansen AP, Komar AA, Zheng X, Merrick WC, Costes S, Lockett SJ, Sonenberg N, Colburn NH (2003) The transformation suppressor Pdcd4 is a novel eukaryotic translation initiation factor 4A binding protein that inhibits translation. Mol Cell Biol 23:26–37. https://doi.org/10.1128/mcb.23.1.26-37.2003
Yang L, Han B, Zhang Z, Wang S, Bai Y, Zhang Y, Tang Y, Du L, Xu L, Wu F, Zuo L, Chen X, Lin Y, Liu K, Ye Q, Chen B, Li B, Tang T, Wang Y, Shen L, Wang G, Ju M, Yuan M, Jiang W, Zhang JH, Hu G, Wang J, Yao H (2020) Extracellular vesicle-mediated delivery of circular RNA SCMH1 promotes functional recovery in rodent and nonhuman primate ischemic stroke models. Circulation 142:556–574. https://doi.org/10.1161/circulationaha.120.045765
Yao X, Yao R, Yi J, Huang F (2019) Upregulation of miR-496 decreases cerebral ischemia/reperfusion injury by negatively regulating BCL2L14. Neurosci Lett 696:197–205. https://doi.org/10.1016/j.neulet.2018.12.039
Zheng T, Shi Y, Zhang J, Peng J, Zhang X, Chen K, Chen Y, Liu L (2019) MiR-130a exerts neuroprotective effects against ischemic stroke through PTEN/PI3K/AKT pathway. Biomed Pharmacother 117:109117. https://doi.org/10.1016/j.biopha.2019.109117
Zheng Y, Zhao P, Lian Y, Li S, Chen Y, Li L (2020) MiR-340-5p alleviates oxygen-glucose deprivation/reoxygenation-induced neuronal injury via PI3K/Akt activation by targeting PDCD4. Neurochem Int 134:104650. https://doi.org/10.1016/j.neuint.2019.104650
Zhou R, Wu Y, Wang W, Su W, Liu Y, Wang Y, Fan C, Li X, Li G, Li Y, Xiong W, Zeng Z (2018) Circular RNAs (circRNAs) in cancer. Cancer Lett 425:134–142. https://doi.org/10.1016/j.canlet.2018.03.035
Zuo L, Li C, Zu J, Yao H, Yan F (2020a) Circular RNA FUNDC1 improves prediction of stroke associated infection in acute ischemic stroke patients with high risk. Biosci Rep 40. https://doi.org/10.1042/bsr20200902
Zuo L, Zhang L, Zu J, Wang Z, Han B, Chen B, Cheng M, Ju M, Li M, Shu G, Yuan M, Jiang W, Chen X, Yan F, Zhang Z, Yao H (2020b) Circulating Circular RNAs as Biomarkers for the Diagnosis and Prediction of Outcomes in Acute Ischemic Stroke. Stroke 51:319–323. https://doi.org/10.1161/strokeaha.119.027348
Zurawska A, Mycko MP, Selmaj KW (2019) Circular RNAs as a novel layer of regulatory mechanism in multiple sclerosis. J Neuroimmunol 334:576971. https://doi.org/10.1016/j.jneuroim.2019.576971
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Conceptualization and Methodology: Jie Liu and Dan Wang; Formal analysis and Data curation: Yu Yao, Xuhua Jiao and Fengjuan Wang; Validation and Investigation: Fengjuan Wang and Jie Liu; Writing - original draft preparation and Writing - review and editing: Fengjuan Wang, Jie Liu and Dan Wang; Approval of final manuscript: all authors.
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Highlight
1. Circ_0007290 is elevated in AIS patients and OGD-induced cell injury model.
2. Knockdown of circ_0007290 alleviates OGD-evoked neuronal injury.
3. Circ_0007290 competitively binds to miR-496 to up-regulate PDCD4 expression.
4. Circ_0007290 knockdown shows protective effects against OGD-evoked neuronal injury via miR-496/PDCD4 axis.
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Wang, F., Liu, J., Wang, D. et al. Knockdown of circ_0007290 alleviates oxygen-glucose deprivation-induced neuronal injury by regulating miR-496/PDCD4 axis. Metab Brain Dis 37, 807–818 (2022). https://doi.org/10.1007/s11011-021-00900-7
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DOI: https://doi.org/10.1007/s11011-021-00900-7