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LncRNA-N1LR Enhances Neuroprotection Against Ischemic Stroke Probably by Inhibiting p53 Phosphorylation

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An Erratum to this article was published on 04 January 2017

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Abstract

In recent years, long noncoding RNAs (lncRNAs) have been shown to have critical roles in a broad range of cell biological processes. However, the activities of lncRNAs during ischemic stroke remain largely unknown. In this study, we carried out a genome-wide lncRNA microarray analysis in rat brains with ischemia/reperfusion (I/R) injury. The results revealed the differential expression of a subset of lncRNAs. Through the construction of lncRNA-mRNA co-expression networks, we identified lncRNA-N1LR as a novel I/R-induced lncRNA. The functions of lncRNA-N1LR were assessed by silencing and overexpressing this lncRNA in vitro and in vivo. We found that lncRNA-N1LR enhanced cell cycle progression and cell proliferation, and inhibited apoptosis in N2a cells subjected to in vitro ischemia (oxygen-glucose deprivation/reoxygenation, OGD/R). Furthermore, we showed that lncRNA-N1LR reduced neuronal apoptosis and neural cell loss in I/R-induced mouse brains. Mechanistically, we discovered that lncRNA-N1LR promoted neuroprotection probably through the inhibition of p53 phosphorylation on serine 15 in a manner that was independent of its location-associated gene Nck1. In summary, our results indicated that lncRNA-N1LR promoted neuroprotection against ischemic stroke probably by inactivating p53. Thus, we propose that lncRNA-N1LR may serve as a potential target for therapeutic intervention following ischemic brain injury.

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  • 04 January 2017

    An erratum to this article has been published.

References

  1. Mozaffarian D, Benjamin EJ, Go AS, Arnett DK, Blaha MJ, Cushman M, Das SR, de Ferranti S et al (2016) Heart disease and stroke statistics-2016 update: a report from the American heart association. Circulation 133(4):e38–e360. doi:10.1161/cir.0000000000000350

    Article  PubMed  Google Scholar 

  2. Mehta SL, Manhas N, Raghubir R (2007) Molecular targets in cerebral ischemia for developing novel therapeutics. Brain Res Rev 54(1):34–66. doi:10.1016/j.brainresrev.2006.11.003

    Article  CAS  PubMed  Google Scholar 

  3. Sleutels F, Zwart R, Barlow DP (2002) The non-coding Air RNA is required for silencing autosomal imprinted genes. Nature 415(6873):810–813. doi:10.1038/415810a

    Article  CAS  PubMed  Google Scholar 

  4. Martianov I, Ramadass A, Serra Barros A, Chow N, Akoulitchev A (2007) Repression of the human dihydrofolate reductase gene by a non-coding interfering transcript. Nature 445(7128):666–670. doi:10.1038/nature05519

    Article  CAS  PubMed  Google Scholar 

  5. Wu Z, Liu X, Liu L, Deng H, Zhang J, Xu Q, Cen B, Ji A (2014) Regulation of lncRNA expression. Cell Mol Biol Lett 19(4):561–575. doi:10.2478/s11658-014-0212-6

    Article  CAS  PubMed  Google Scholar 

  6. Yang F, Zhang L, Huo XS, Yuan JH, Xu D, Yuan SX, Zhu N, Zhou WP et al (2011) Long noncoding RNA high expression in hepatocellular carcinoma facilitates tumor growth through enhancer of zeste homolog 2 in humans. Hepatology 54(5):1679–1689. doi:10.1002/hep.24563

    Article  CAS  PubMed  Google Scholar 

  7. Wan G, Hu X, Liu Y, Han C, Sood AK, Calin GA, Zhang X, Lu X (2013) A novel non-coding RNA lncRNA-JADE connects DNA damage signalling to histone H4 acetylation. EMBO J 32(21):2833–2847. doi:10.1038/emboj.2013.221

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Wu P, Zuo X, Deng H, Liu X, Liu L, Ji A (2013) Roles of long noncoding RNAs in brain development, functional diversification and neurodegenerative diseases. Brain Res Bull 97:69–80. doi:10.1016/j.brainresbull.2013.06.001

    Article  CAS  PubMed  Google Scholar 

  9. Dharap A, Nakka VP, Vemuganti R (2012) Effect of focal ischemia on long noncoding RNAs. Stroke; A Journal of Cerebral Circulation 43(10):2800–2802. doi:10.1161/STROKEAHA.112.669465

    Article  CAS  PubMed Central  Google Scholar 

  10. Begum G, Yuan H, Kahle KT, Li L, Wang S, Shi Y, Shmukler BE, Yang SS et al (2015) Inhibition of WNK3 kinase signaling reduces brain damage and accelerates neurological recovery after stroke. Stroke; A Journal of Cerebral Circulation 46(7):1956–1965. doi:10.1161/strokeaha.115.008939

    Article  CAS  PubMed Central  Google Scholar 

  11. Chi W, Meng F, Li Y, Wang Q, Wang G, Han S, Wang P, Li J (2014) Downregulation of miRNA-134 protects neural cells against ischemic injury in N2A cells and mouse brain with ischemic stroke by targeting HSPA12B. Neuroscience 277:111–122. doi:10.1016/j.neuroscience.2014.06.062

    Article  CAS  PubMed  Google Scholar 

  12. Tatlisumak T, Takano K, Carano RA, Fisher M (1996) Effect of basic fibroblast growth factor on experimental focal ischemia studied by diffusion-weighted and perfusion imaging. Stroke; A Journal of Cerebral Circulation 27(12):2292–2297 discussion 2298

    Article  CAS  Google Scholar 

  13. Ashwal S, Tone B, Tian HR, Cole DJ, Liwnicz BH, Pearce WJ (1999) Core and penumbral nitric oxide synthase activity during cerebral ischemia and reperfusion in the rat pup. Pediatr Res 46(4):390–400. doi:10.1203/00006450-199910000-00006

    Article  CAS  PubMed  Google Scholar 

  14. Chen J, Deng F, Singh SV, Wang QJ (2008) Protein kinase D3 (PKD3) contributes to prostate cancer cell growth and survival through a PKCepsilon/PKD3 pathway downstream of Akt and ERK 1/2. Cancer Res 68(10):3844–3853. doi:10.1158/0008-5472.can-07-5156

    Article  CAS  PubMed  Google Scholar 

  15. Nayak RR, Kearns M, Spielman RS, Cheung VG (2009) Coexpression network based on natural variation in human gene expression reveals gene interactions and functions. Genome Res 19(11):1953–1962. doi:10.1101/gr.097600.109

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  16. DiFiglia M, Sena-Esteves M, Chase K, Sapp E, Pfister E, Sass M, Yoder J, Reeves P et al (2007) Therapeutic silencing of mutant huntingtin with siRNA attenuates striatal and cortical neuropathology and behavioral deficits. Proc Natl Acad Sci U S A 104(43):17204–17209. doi:10.1073/pnas.0708285104

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  17. Dubrac A, Genet G, Ola R, Zhang F, Pibouin-Fragner L, Han J, Zhang J, Thomas JL et al (2016) Targeting NCK-mediated endothelial cell front-rear polarity inhibits neovascularization. Circulation 133(4):409–421. doi:10.1161/circulationaha.115.017537

    Article  PubMed  Google Scholar 

  18. Chen J, Leskov IL, Yurdagul A Jr, Thiel B, Kevil CG, Stokes KY, Orr AW (2015) Recruitment of the adaptor protein Nck to PECAM-1 couples oxidative stress to canonical NF-kappaB signaling and inflammation. Sci Signal 8(365):ra20. doi:10.1126/scisignal.2005648

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  19. Leker RR, Aharonowiz M, Greig NH, Ovadia H (2004) The role of p53-induced apoptosis in cerebral ischemia: effects of the p53 inhibitor pifithrin alpha. Exp Neurol 187(2):478–486. doi:10.1016/j.expneurol.2004.01.030

    Article  CAS  PubMed  Google Scholar 

  20. Maeda K, Hata R, Gillardon F, Hossmann KA (2001) Aggravation of brain injury after transient focal ischemia in p53-deficient mice. Brain Res Mol Brain Res 88(1–2):54–61

    Article  CAS  PubMed  Google Scholar 

  21. Culmsee C, Zhu X, Yu QS, Chan SL, Camandola S, Guo Z, Greig NH, Mattson MP (2001) A synthetic inhibitor of p53 protects neurons against death induced by ischemic and excitotoxic insults, and amyloid beta-peptide. J Neurochem 77(1):220–228

    Article  CAS  PubMed  Google Scholar 

  22. Chen H, Yoshioka H, Kim GS, Jung JE, Okami N, Sakata H, Maier CM, Narasimhan P et al (2011) Oxidative stress in ischemic brain damage: mechanisms of cell death and potential molecular targets for neuroprotection. Antioxid Redox Signal 14(8):1505–1517. doi:10.1089/ars.2010.3576

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Crumrine RC, Thomas AL, Morgan PF (1994) Attenuation of p53 expression protects against focal ischemic damage in transgenic mice. J Cereb Blood Flow Metab 14(6):887–891. doi:10.1038/jcbfm.1994.119

    Article  CAS  PubMed  Google Scholar 

  24. Yin KJ, Deng Z, Hamblin M, Xiang Y, Huang H, Zhang J, Jiang X, Wang Y et al (2010) Peroxisome proliferator-activated receptor delta regulation of miR-15a in ischemia-induced cerebral vascular endothelial injury. J Neurosci 30(18):6398–6408. doi:10.1523/jneurosci.0780-10.2010

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Yin KJ, Deng Z, Huang H, Hamblin M, Xie C, Zhang J, Chen YE (2010) miR-497 regulates neuronal death in mouse brain after transient focal cerebral ischemia. Neurobiol Dis 38(1):17–26. doi:10.1016/j.nbd.2009.12.021

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  26. Harraz MM, Eacker SM, Wang X, Dawson TM, Dawson VL (2012) MicroRNA-223 is neuroprotective by targeting glutamate receptors. Proc Natl Acad Sci U S A 109(46):18962–18967. doi:10.1073/pnas.1121288109

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Martens JA, Laprade L, Winston F (2004) Intergenic transcription is required to repress the Saccharomyces cerevisiae SER3 gene. Nature 429(6991):571–574. doi:10.1038/nature02538

    Article  CAS  PubMed  Google Scholar 

  28. Doyle KP, Simon RP, Stenzel-Poore MP (2008) Mechanisms of ischemic brain damage. Neuropharmacology 55(3):310–318. doi:10.1016/j.neuropharm.2008.01.005

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. Errington TM, Macara IG (2013) Depletion of the adaptor protein NCK increases UV-induced p53 phosphorylation and promotes apoptosis. PLoS One 8(9):e76204. doi:10.1371/journal.pone.0076204

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  30. Banasiak KJ, Haddad GG (1998) Hypoxia-induced apoptosis: effect of hypoxic severity and role of p53 in neuronal cell death. Brain Res 797(2):295–304

    Article  CAS  PubMed  Google Scholar 

  31. Banin S, Moyal L, Shieh S, Taya Y, Anderson CW, Chessa L, Smorodinsky NI, Prives C et al (1998) Enhanced phosphorylation of p53 by ATM in response to DNA damage. Science 281(5383):1674–1677

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

We thank the Guangdong Provincial Key Laboratory of Malignant Tumor Epigenetics and Gene Regulation for flow cytometry support. This work was supported by the National Nature Science Foundation of China grant NSFC81370449.

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Authors and Affiliations

  1. Center for Drug Research and Development, Zhujiang Hospital, Southern Medical University, Industrial avenue253, Guangzhou, 510282, China

    Zhuomin Wu, Yixin Qin, Qian Xu, Shuai He, Bohong Cen, Wenjie Liao & Aimin Ji

  2. Pharmacy Department, Chengdu First People’s Hospital/ Chengdu Integrated TCM & Western Medicine Hospital, Chengdu, China

    Ping Wu

  3. Institute of Neurosciences and the Second Affiliated Hospital of Guangzhou Medical University, Guangzhou, China

    Xialin Zuo

  4. The Second Clinical College of Southern Medical University, Guangzhou, China

    Na Yu

Authors
  1. Zhuomin Wu
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  2. Ping Wu
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Correspondence to Aimin Ji.

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The authors declare that they have no conflict of interest.

Additional information

Zhuomin Wu and Ping Wu contributed equally to this work.

An erratum to this article is available at https://doi.org/10.1007/s12035-016-0354-9.

Electronic supplementary material

Table S1

Sequences of primers and siRNA used in this study. (DOCX 19 kb)

Table S2

Differentially expressed lncRNAs identified in ischemic penumbra tissue. (PDF 490 kb)

Table S3

Differentially expressed mRNAs identified in ischemic penumbra tissue. (PDF 787 kb)

Table S4

The functions of mRNAs up-regulated over 100 fold-changes in ischemic penumbra. (PDF 221 kb)

Table S5

Functions of genes connected to lncRNA-N1LR in sub-network. (PDF 48 kb)

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Wu, Z., Wu, P., Zuo, X. et al. LncRNA-N1LR Enhances Neuroprotection Against Ischemic Stroke Probably by Inhibiting p53 Phosphorylation. Mol Neurobiol 54, 7670–7685 (2017). https://doi.org/10.1007/s12035-016-0246-z

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