The Neuroprotective Efficacy of Cell-Penetrating Peptides TAT, Penetratin, Arg-9, and Pep-1 in Glutamic Acid, Kainic Acid, and In Vitro Ischemia Injury Models Using Primary Cortical Neuronal Cultures

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• 作者：Bruno P. Meloni (1) (2)
  Amanda J. Craig (1)
  Nadia Milech (3)
  Richard M. Hopkins (3)
  Paul M. Watt (3)
  Neville W. Knuckey (1) (2)
  
• 关键词：Cell ; penetrating peptides ; TAT ; Arg ; 9 ; Penetratin ; Pep ; 1 ; Excitotoxicity ; In vitro ischemia
• 刊名：Cellular and Molecular Neurobiology
• 出版年：2014
• 出版时间：March 2014
• 年：2014
• 卷：34
• 期：2
• 页码：173-181
• 全文大小：345 KB
• 参考文献：1. Aarts M, Liu Y, Liu L, Besshoh S, Arundine M, Gurd JW, Wang YT, Salter MW, Tymianski M (2002) Treatment of ischemic brain damage by perturbing NMDA receptor-PSD-95 protein interactions. Science 298:846-50 CrossRef
  2. ARAMIS (2011) Project number: 8909.1. http://www.aramis.admin.ch. Accessed 2 Nov 2013
  3. Arthur PG, Matich GP, Pang WW, Yu DY, Bogoyevitch MA (2007) Necrotic death of neurons following an excitotoxic insult is prevented by a peptide inhibitor of c-jun N-terminal kinase. J Neurochem 102:65-6 j.1471-4159.2007.04618.x" target="_blank" title="It opens in new window">CrossRef
  4. Ashpole NM, Hudmon A (2011) Excitotoxic neuroprotection and vulnerability with CaMKII inhibition. Mol Cell Neurosci 46:720-30 j.mcn.2011.02.003" target="_blank" title="It opens in new window">CrossRef
  5. Borsello T, Clarke PG, Hirt L, Vercelli A, Repici M, Schorderet DF, Bogousslavsky J, Bonny C (2003) A peptide inhibitor of c-Jun N-terminal kinase protects against excitotoxicity and cerebral ischemia. Nat Med 9:1180-186 CrossRef
  6. Cardozo AK, Buchillier V, Mathieu M, Chen J, Ortis F, Ladrière L, Allaman-Pillet N, Poirot O, Kellenberger S, Beckmann JS, Eizirik DL, Bonny C, Maurer F (2007) Cell-permeable peptides induce dose-and length-dependent cytotoxic effects. Biochim Biophys Acta 1768:2222-234 j.bbamem.2007.06.003" target="_blank" title="It opens in new window">CrossRef
  7. Colombo A, Repici M, Pesaresi M, Santambrogio S, Forloni G, Borsello T (2007) The TAT-JNK inhibitor peptide interferes with beta amyloid protein stability. Cell Death Differ 14:1845-848 j.cdd.4402202" target="_blank" title="It opens in new window">CrossRef
  8. Craig AJ, Meloni BP, Watt PM, Knuckey NW (2011) Attenuation of neuronal death by peptide inhibitors of AP-1 activation in acute and delayed in vitro ischaemia (oxygen/glucose deprivation) models. Int J Pept Res Ther 17:1- CrossRef
  9. Doeppner TR, Nagel F, Dietz GPH, Weise J, T?nges L, Schwarting S, B?hr M (2009) TAT-HSP70-mediated neuroprotection and increased survival of neuronal precursor cells after focal cerebral ischemia in mice. J Cereb Blood Flow Metab 29:1187-196 jcbfm.2009.44" target="_blank" title="It opens in new window">CrossRef
  10. Dolgin E (2012) To serve and neuroprotect. Nat Med 18:1003-006 CrossRef
  11. Ferrer-Montiel AV, Merino JM, Blondelle SE, Perez-Payà E, Houghten RA, Montal M (1998) Selected peptides targeted to the NMDA receptor channel protect neurons from excitotoxic death. Nat Biotechnol 16:286-91 CrossRef
  12. Frankel AD, Pabo CO (1988) Cellular uptake of the TAT protein from human immunodeficiency virus. Cell 55:1189-193 CrossRef
  13. Green M, Loewenstein PM (1988) Autonomous functional domains of chemically synthesized human immunodeficiency virus TAT trans-activator protein. Cell 55:1179-188 CrossRef
  14. Hirose H, Takeuchi T, Osakada H, Pujals S, Katayama S, Nakase I, Kobayashi S, Haraguchi T, Futaki S (2012) Transient focal membrane deformation induced by arginine-rich peptides leads to their direct penetration into cells. Mol Ther 20:984-93 CrossRef
  15. Ho A, Schwarze SR, Mermelstein SJ, Waksman G, Dowdy SF (2001) Synthetic protein transduction domains: enhanced transduction potential in vitro and in vivo. Cancer Res 61:474-77
  16. Kacprzak MM, Peinado JR, Than ME, Appel J, Henrich S, Lipkind G, Houghten RA, Bode W, Lindberg I (2004) Inhibition of furin by polyarginine-containing peptides: nanomolar inhibition by nona-d -arginine. J Biol Chem 279:36788-6794 jbc.M400484200" target="_blank" title="It opens in new window">CrossRef
  17. Kilic ü, Kilic E, Dietz GPH, B?hr M (2003) Intravenous TAT-GDNF is protective after focal cerebral ischemia in mice. Stroke 34:1304-310 CrossRef
  18. Lai Y, Du L, Dunsmore KE, Jenkins LW, Wong HR, Clark RS (2005) Selectively increasing inducible heat shock protein 70 via TAT-protein transduction protects neurons from nitrosative stress and excitotoxicity. J Neurochem 94:360-66 j.1471-4159.2005.03212.x" target="_blank" title="It opens in new window">CrossRef
  19. Li W, Huang Y, Reid R, Steiner J, Malpica-Llanos T, Darden TA, Shankar SK, Mahadevan A, Satishchandra P, Nath A (2008) NMDA receptor activation by HIV-Tat protein is clade dependent. J Neurosci 28:12190-2198
  20. Liu XM, Pei DS, Guan QH, Sun YF, Wang XT, Zhang QX, Zhang GY (2006) Neuroprotection of TAT-GluR6-9c against neuronal death induced by kainate in rat hippocampus via nuclear and non-nuclear pathways. J Biol Chem 281:17432-7445 jbc.M513490200" target="_blank" title="It opens in new window">CrossRef
  21. Meade AJ, Meloni BP, Mastaglia FL, Knuckey NW (2009) The application of cell penetrating peptides for the delivery of neuroprotective peptides/proteins in experimental cerebral ischemia studies. J Exp Stroke Transl Med 2:22-0 CrossRef
  22. Meade AJ, Meloni BP, Mastaglia FL, Watt PM, Knuckey NW (2010a) AP-1 inhibitory peptides attenuate in vitro cortical neuronal cell death induced by kainic acid. Brain Res 1360:8-6 j.brainres.2010.09.007" target="_blank" title="It opens in new window">CrossRef
  23. Meade AJ, Meloni BP, Cross J, Bakker AJ, Fear MW, Mastaglia FL, Watt PM, Knuckey NW (2010b) AP-1 inhibitory peptides are neuroprotective following acute glutamate excitotoxicity in primary cortical neuronal cultures. J Neurochem 112:258-70 j.1471-4159.2009.06459.x" target="_blank" title="It opens in new window">CrossRef
  24. Meloni BP, Majda BT, Knuckey NW (2001) Establishment of neuronal in vitro models of ischemia in 96-well microtiter strip-plates that result in acute, progressive and delayed neuronal death. Neuroscience 108:17-6 CrossRef
  25. Meloni BP, Meade AJ, Kitikomolsuk D, Knuckey NW (2011) Characterisation of neuronal cell death in acute and delayed in vitro ischemia (oxygen/glucose deprivation) models. J Neurosci Methods 195:67-4 j.jneumeth.2010.11.023" target="_blank" title="It opens in new window">CrossRef
  26. Milletti F (2012) Cell-penetrating peptides: classes, origin, and current landscape. Drug Discov Today 17:850-60 j.drudis.2012.03.002" target="_blank" title="It opens in new window">CrossRef
  27. Moschos SA, Jones SW, Perry MM, Williams AE, Erjefalt JS, Turner JJ, Barnes PJ, Sproat BS, Gait MJ, Lindsay MA (2007) Lung delivery studies using siRNA conjugated to TAT(48-0) and penetratin reveal peptide induced reduction in gene expression and induction of innate immunity. Bioconjug Chem 18:1450-459 CrossRef
  28. Nagel F, Falkenburger BH, T?nges L, Kowsky S, P?ppelmeyer C, Schulz JB, B?hr M, Dietz GP (2008) TAT-HSP70 protects dopaminergic neurons in midbrain cultures and in the substantia nigra in models of Parkinson’s disease. J Neurochem 105:853-64 j.1471-4159.2007.05204.x" target="_blank" title="It opens in new window">CrossRef
  29. Nath A, Psooy K, Martin C, Knudsen B, Magnuson DS, Haughey N, Geiger JD (1996) Identification of a human immunodeficiency virus type 1 TAT epitope that is neuroexcitatory and neurotoxic. J Virol 70:1475-480
  30. Palm-Apergi C, L?nn P, Dowdy SF (2012) Do cell-penetrating peptides actually “penetrate-cellular membranes? Mol Ther 20:695-97 CrossRef
  31. Vaslin A, Rummel C, Clarke PG (2009) Unconjugated TAT carrier peptide protects against excitotoxicity. Neurotox Res 15:123-26 CrossRef
  32. Xu W, Zhou M, Baudry M (2008) Neuroprotection by cell permeable TAT-mGluR1 peptide in ischemia: synergy between carrier and cargo sequences. Neuroscientist 14:409-14 CrossRef
  33. Zhang S, Taghibiglou C, Girling K, Dong Z, Lin SZ, Lee W, Shyu WC, Wang YT (2013) Critical role of increased PTEN nuclear translocation in excitotoxic and ischemic neuronal injuries. J Neurosci 33:7997-008 CrossRef
  
• 作者单位：Bruno P. Meloni (1) (2)
  Amanda J. Craig (1)
  Nadia Milech (3)
  Richard M. Hopkins (3)
  Paul M. Watt (3)
  Neville W. Knuckey (1) (2)
  
  1. Centre for Neuromuscular and Neurological Disorders, The University of Western Australia and Australian Neuromuscular Research Institute, A Block, 4th Floor, QEII Medical Centre, Verdun St, Nedlands, WA, 6009, Australia
  2. Department of Neurosurgery, Sir Charles Gairdner Hospital, Nedlands, WA, Australia
  3. Phylogica Ltd and Telethon Institute for Child Health Research, The University of Western Australia, Nedlands, WA, Australia
  
• ISSN：1573-6830

文摘

Cell-penetrating peptides (CPPs) are small peptides (typically 5-5 amino acids), which are used to facilitate the delivery of normally non-permeable cargos such as other peptides, proteins, nucleic acids, or drugs into cells. However, several recent studies have demonstrated that the TAT CPP has neuroprotective properties. Therefore, in this study, we assessed the TAT and three other CPPs (penetratin, Arg-9, Pep-1) for their neuroprotective properties in cortical neuronal cultures following exposure to glutamic acid, kainic acid, or in vitro ischemia (oxygen–glucose deprivation). Arg-9, penetratin, and TAT-D displayed consistent and high level neuroprotective activity in both the glutamic acid (IC50: 0.78, 3.4, 13.9?μM) and kainic acid (IC50: 0.81, 2.0, 6.2?μM) injury models, while Pep-1 was ineffective. The TAT-D isoform displayed similar efficacy to the TAT-L isoform in the glutamic acid model. Interestingly, Arg-9 was the only CPP that displayed efficacy when washed-out prior to glutamic acid exposure. Neuroprotection following in vitro ischemia was more variable with all peptides providing some level of neuroprotection (IC50; Arg-9: 6.0?μM, TAT-D: 7.1?μM, penetratin/Pep-1: >10?μM). The positive control peptides JNKI-1D-TAT (JNK inhibitory peptide) and/or PYC36L-TAT (AP-1 inhibitory peptide) were neuroprotective in all models. Finally, in a post-glutamic acid treatment experiment, Arg-9 was highly effective when added immediately after, and mildly effective when added 15?min post-insult, while the JNKI-1D-TAT control peptide was ineffective when added post-insult. These findings demonstrate that different CPPs have the ability to inhibit neurodamaging events/pathways associated with excitotoxic and ischemic injuries. More importantly, they highlight the need to interpret neuroprotection studies when using CPPs as delivery agents with caution. On a positive note, the cytoprotective properties of CPPs suggests they are ideal carrier molecules to deliver neuroprotective drugs to the CNS following injury and/or potential neuroprotectants in their own right.