Skip to main content

Advertisement

SpringerLink
Log in

Combined Gene Therapy to Reduce the Neuronal Damage in the Mouse Model of Focal Ischemic Injury

  • Published:
Journal of Molecular Neuroscience Aims and scope Submit manuscript

Abstract

Research into stroke is driven by frustration over the limited available therapeutics. Targeting a single aspect of this multifactorial disease contributes to the therapeutic boundaries. To overcome this, we devised a novel multifactorial-cocktail treatment, using lentiviruses encoding excitatory amino acid transporter 2 (EAAT2(, glutamate dehydrogenase 2 (GDH2), and nuclear factor E2-related factor 2 (Nrf2) genes, that acts synergistically to address the effected excito-oxidative axis. Here, we used the vasoconstrictor endothelin-1 (ET-1) to induce focal ischemic injury in mice by direct injection into the striatum. Mice treated with the mixture of these three genes show significant improvement in body balance, motor coordination, and decreased motor asymmetry compared to each gene separately. These results demonstrate that overexpression of the combined EAAT2, GDH2, and NRF2 genes can provide neuroprotection after ischemic injury.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Anderson CS, Linto J, Stewart-Wynne EG (1995) A population-based assessment of the impact and burden of caregiving for long-term stroke survivors. Stroke 26:843–849

    Article  CAS  PubMed  Google Scholar 

  • Anderson MF, Blomstrand F, Blomstrand C, Eriksson PS, Nilsson M (2003) Astrocytes and stroke: networking for survival? Neurochem Res 28:293–305

    Article  CAS  PubMed  Google Scholar 

  • Barone FC, Feuerstein GZ (1999) Inflammatory mediators and stroke: new opportunities for novel therapeutics. J Cereb Blood Flow Metab 19:819–834

    Article  CAS  PubMed  Google Scholar 

  • Begum G, Song S, Wang S, Zhao H, Bhuiyan MIH, Li E, Nepomuceno R, Ye Q, Sun M, Calderon MJ, Stolz DB, St. Croix C, Watkins SC, Chen Y, He P, Shull GE, Sun D (2018) Selective knockout of astrocytic Na1/H1 exchanger isoform 1 reduces astrogliosis, BBB damage, infarction, and improves neurological function after ischemic stroke. Glia 66:126–144

    Article  PubMed  Google Scholar 

  • Benkler C, Barhum Y, Ben-Zur T, Offen D (2016) Multifactorial gene therapy enhancing the glutamate uptake system and reducing oxidative stress delays symptom onset and prolongs survival in the SOD1-G93A ALS mouse model. J Mol Neurosci 58:46–58

    Article  CAS  PubMed  Google Scholar 

  • Chamorro Á, Dirnagl U, Urra X, Planas AM (2016) Neuroprotection in acute stroke: targeting excitotoxicity, oxidative and nitrosative stress, and inflammation. Lancet Neurol 15:869–881

    Article  CAS  PubMed  Google Scholar 

  • Chaudhry FA, Lehre KP, van Lookeren et al (1995) Glutamate transporters in glial plasma membranes: highly differentiated localizations revealed by quantitative ultrastructural immunocytochemistry. Neuron 15:711–720

    Article  CAS  PubMed  Google Scholar 

  • Choi K, Kim J, Kim GW, Choi C (2009) Oxidative stress-induced necrotic cell death via mitochondria-dependent burst of reactive oxygen species. Curr Neurovasc Res 6:213–322

    Article  CAS  PubMed  Google Scholar 

  • Chu K, Lee ST, Sinn DI (2007) Pharmacological induction of ischemic tolerance by glutamate transporter-1 (EAAT2) upregulation. Stroke 38:177–182

    Article  CAS  PubMed  Google Scholar 

  • Cuzzocrea S, Riley DP, Caputi AP, Salvemini D (2001) Antioxidant therapy: a new pharmacological approach in shock, inflammation, and ischemia/reperfusion injury. Pharmacol Rev 53:135–159

    CAS  PubMed  Google Scholar 

  • Dirnagl U, Iadecola C, Moskowitz MA (1999a) Pathobiology of ischaemic stroke: an integrated view. Trends Neurosci 22:391–397

    Article  CAS  PubMed  Google Scholar 

  • Dirnagl U, Iadecola C, Moskowitz M (1999b) Pathobiology of ischaemic stroke: an integrated view. 4441. Trends Neurosci 22:391–397

    Article  CAS  PubMed  Google Scholar 

  • Donnan GA, Davis SM (2008) Neuroprotection: still achievable in humans. Stroke 39:525–530

    Article  PubMed  Google Scholar 

  • Fuxe K, Cintra A, Andbjer B (1989) Centrally administered endothelin-1 produces lesions in the brain of the male rat. Acta Physiol Scand 137:155–156

    Article  CAS  PubMed  Google Scholar 

  • Glat MJ, Benninger F, Barhum Y, Ben-Zur T, Kogan E, Steiner I, Yaffe D, Offen D (2016) Ectopic muscle expression of neurotrophic factors improves recovery after nerve injury. J Mol Neurosci 58(1):39–45

    Article  CAS  PubMed  Google Scholar 

  • Han B, Haley WE (1999) Family caregiving for patients with stroke: review and analysis. Stroke 30:1478–1485

    Article  CAS  PubMed  Google Scholar 

  • Harvey BK, Airavaara M, Hinzman J, Wires EM, Chiocco MJ, Howard DB, Shen H, Gerhardt G, Hoffer BJ, Wang Y (2011) Targeted over-expression of glutamate transporter 1 (GLT-1) reduces ischemic brain injury in a rat model of stroke. PLoS One 6(8). https://doi.org/10.1371/journal.pone.0022135

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Horie N, Maag AL, Hamilton SA, Shichinohe H, Bliss TM, Steinberg GK (2008) Mouse model of focal cerebral ischemia using endothelin-1. J Neurosci Methods 173:286–290

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Hudson RC, Daniel RM (1993) L-glutamate dehydrogenases: distribution, properties and mechanism. Comparative Biochemistry and Physiology. B, Comparative Biochemistry 106:767–792

    Article  CAS  PubMed  Google Scholar 

  • Ishii T, Itoh K, Takahashi S, Sato H, Yanagawa T, Katoh Y, Bannai S, Yamamoto M (2000) Transcription factor Nrf2 coordinately regulates a group of oxidative stress-inducible genes in macrophages. J Biol Chem 275:16023–16029

    Article  CAS  PubMed  Google Scholar 

  • Itoh K, Chiba T, Takahashi S, Ishii T, Igarashi K, Katoh Y, Oyake T, Hayashi N, Satoh K, Hatayama I, Yamamoto M, Nabeshima Y (1997) An Nrf2/small Maf heterodimer mediates the induction of phase II detoxifying enzyme genes through antioxidant response elements. Biochem Biophys Res Commun 236(2):313–322

    Article  CAS  PubMed  Google Scholar 

  • Kurosawa M, Fuxe K, Hallström A, Goiny M, Cintra A, Ungerstedt U (1991) Responses of blood flow, extracellular lactate, and dopamine in the striatum to intrastriatal injection of endothelin-1 in anesthetized rats. J Cardiovasc Pharmacol 17:S340–S342

    Article  CAS  PubMed  Google Scholar 

  • Lai TW, Zhang S, Wang YT (2014) Excitotoxicity and stroke: identifying novel targets for neuroprotection. Prog Neurobiol 115:157–188

    Article  CAS  PubMed  Google Scholar 

  • Lakhan SE, Kirchgessner A, Hofer M (2009a) Inflammatory mechanisms in ischemic stroke: therapeutic approaches. J Transl Med 7:97–102

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Lakhan SE, Kirchgessner A, Hofer M (2009b) Inflammatory mechanisms in ischemic stroke: therapeutic approaches. J Transl Med 7:97–103

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Laterza C, Wattananit S, Uoshima N, Ge R, Pekny R, Tornero D, Monni E, Lindvall O, Kokaia Z (2017) Monocyte depletion early after stroke promotes neurogenesis from endogenous neural stem cells in the adult brain. Exp Neurol 297:129–137

    Article  CAS  PubMed  Google Scholar 

  • Li M, Li Z, Yao Y, Jin WN, Wood K, Liu Q, Shi FD, Hao J (2017) Astrocyte-derived interleukin-15 exacerbates ischemic brain injury via propagation of cellular immunity. Proc Natl Acad Sci U S A 114:E396–E405

    Article  CAS  PubMed  Google Scholar 

  • Lin CL, Kong Q, Cuny GD, Glicksman MA (2012) Glutamate transporter EAAT2: a new target for the treatment of neurodegenerative diseases. Future Med Chem 4:1689–1700

    Article  CAS  PubMed  Google Scholar 

  • Lo EH, Dalkara T, Moskowitz MA (2003) Neurological diseases: mechanisms, challenges and opportunities in stroke. Nat Rev Neurosci 4:399–414

    Article  CAS  PubMed  Google Scholar 

  • Matsui T, Mori T, Tateishi N, Kagamiishi Y, Satoh S, Katsube N, Morikawa E, Morimoto T, Ikuta F, Asano T (2002) Astrocytic activation and delayed infarct expansion after permanent focal ischemia in rats. Part I: enhanced astrocytic synthesis of S-100?? In the periinfarct area precedes delayed infarct expansion. J Cereb Blood Flow Metab 22:711–722

    Article  CAS  PubMed  Google Scholar 

  • Muir KW, Tyrrell P, Sattar N, Warburton E (2007) Inflammation and ischaemic stroke. Curr Opin Neurol 20:334–342

    Article  CAS  PubMed  Google Scholar 

  • Murphy S (2000) Production of nitric oxide by glial cells: regulation and potential roles in the CNS. Glia 29:1–13

    Article  CAS  PubMed  Google Scholar 

  • Nguemeni C, Gomez-Smith M, Jeffers MS, Schuch CP, Corbett D (2015) Time course of neuronal death following endothelin-1 induced focal ischemia in rats. J Neurosci Methods 242:72–76

    Article  CAS  PubMed  Google Scholar 

  • Nguyen T, Nioi P, Picket CB (2009) The Nrf2-antioxidant response element signaling pathway and its activation by oxidative stress. J Biol Chem 284:13291–13295

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Plaitakis A, Metaxari M, Shashidharan P (2000) Nerve tissue-specific (GLUD 2) and housekeeping (GLUD 1) human glutamate dehydrogenases are regulated by distinct allosteric mechanisms : implications for biologic function. J Neurochem 75(5):1862–1869

  • Plaitakis A, Metaxari M, Shashidharan P (2010) Gain-of-function variant in GLUD2 glutamate dehydrogenase modifies Parkinson’s disease onset. Eur J Hum Genet 18:336–341

    Article  CAS  PubMed  Google Scholar 

  • Rao VL, Dogan A, Bowen KK, Todd KG, Dempsey RJ (2001) Antisense knockdown of the glial glutamate transporter GLT-1 exacerbates hippocampal neuronal damage following traumatic injury to rat brain. Eur J Neurosci 13:119–128

    CAS  PubMed  Google Scholar 

  • Rothstein JD, Dykes-Hoberg M, Pardo CA, Bristol LA, Jin L, Kuncl RW, Kanai Y, Hediger MA, Wang Y, Schielke JP, Welty DF (1996) Knockout of glutamate transporters reveals a major role for astroglial transport in excitotoxicity and clearance of glutamate. Neuron 16:675–686

    Article  CAS  PubMed  Google Scholar 

  • Schallert T, Fleming SM, Leasure JL, Tillerson JL, Bland ST (2000) CNS plasticity and assessment of forelimb sensorimotor outcome in unilateral rat models of stroke, cortical ablation, parkinsonism and spinal cord injury. Neuropharmacology 39:777–787

    Article  CAS  PubMed  Google Scholar 

  • Shashidharan P, Clarke DD, Ahmed N, Moschonas N, Plaitakis A (1997) Nerve tissue-specific human glutamate dehydrogenase that is thermolabile and highly regulated by ADP. J Neurochem 68:1804–1811

    Article  CAS  PubMed  Google Scholar 

  • Shih AY, Li P, Murphy TH (2005) A small-molecule-inducible Nrf2-mediated antioxidant response provides effective prophylaxis against cerebral ischemia in vivo. J Neurosci 25:10321–10335

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Stineman MG, Maislin G, Fiedler RC, Granger CV (1997) A prediction model for functional recovery in stroke. Stroke 28:550–556

    Article  CAS  PubMed  Google Scholar 

  • Surh YJ, Kundu JK, Na HK (2008) Nrf2 as a master redox switch in turning on the cellular signaling involved in the induction of cytoprotective genes by some chemopreventive phytochemicals. Planta Med 74(13):1526–1539

    Article  CAS  PubMed  Google Scholar 

  • Takagi T, Kitashoji A, Iwawaki T, Tsuruma K, Shimazawa M, Yoshimura S, Iwama T, Hara H (2014) Temporal activation of Nrf2 in the penumbra and Nrf2 activator-mediated neuroprotection in ischemia-reperfusion injury. Free Radic Biol Med 72:124–133

    Article  CAS  PubMed  Google Scholar 

  • Takahashi K, Foster JB, Lin C (2015) Glutamate transporter EAAT2: regulation, function, and potential as a therapeutic target for neurological and psychiatric disease. Cell Mol Life Sci 72:3489–3506

    Article  CAS  PubMed  Google Scholar 

  • Wang Q, Tang XN, Yenari MA (2007) The inflammatory response in stroke. J Neuroimmunol 184:53–68

    Article  CAS  PubMed  Google Scholar 

  • Weller ML, Stone IM, Goss A, Rau T, Rova C, Poulsen DJ (2008) Selective overexpression of excitatory amino acid transporter 2 (EAAT2) in astrocytes enhances neuroprotection from moderate but not severe hypoxia-ischemia. Neuroscience 155(4):1204–1211

    Article  CAS  PubMed  Google Scholar 

  • Yamauchi K, Nakano Y, Imai T, Takagi T, Tsuruma K, Shimazawa M, Iwama T, Hara H (2016) A novel nuclear factor erythroid 2-related factor 2 (Nrf2) activator RS9 attenuates brain injury after ischemia reperfusion in mice. Neuroscience 333:302–310

    Article  CAS  PubMed  Google Scholar 

  • Yang C, Zhang X, Fan H, Liu Y (2009) Curcumin upregulates transcription factor Nrf2, HO-1 expression and protects rat brains against focal ischemia. Brain Res 1282:133–141

    Article  CAS  PubMed  Google Scholar 

  • Zhao J, Kobori N, Aronowski J, Dash PK (2006) Sulforaphane reduces infarct volume following focal cerebral ischemia in rodents. Neurosci Lett 393(2–3):108–112

    Article  CAS  PubMed  Google Scholar 

Download references

Funding

This study received funds as a scholarship from Mr. Martin Davis to LM.

Author information

Authors and Affiliations

  1. Sagol School of Neuroscience, Tel-Aviv University, Tel Aviv, Israel

    Lior Molcho, Mica Glat & Daniel Offen

  2. Felsenstein Medical Research Center, Sackler School of Medicine, Tel-Aviv University, Tel Aviv, Israel

    Tali Ben-Zur, Yael Barhum, Ariel Angel & Daniel Offen

Authors
  1. Lior Molcho
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Tali Ben-Zur
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yael Barhum
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Ariel Angel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Mica Glat
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Daniel Offen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Daniel Offen.

Ethics declarations

Competing Interests

DO holds several patents related to gene therapy in neurodegenerative diseases. All were assigned to “Ramot at Tel Aviv University.” DO is a consultant to “Brainstorm Cell Therapeutics.” The other authors have nothing to disclose.

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Molcho, L., Ben-Zur, T., Barhum, Y. et al. Combined Gene Therapy to Reduce the Neuronal Damage in the Mouse Model of Focal Ischemic Injury. J Mol Neurosci 66, 180–187 (2018). https://doi.org/10.1007/s12031-018-1143-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12031-018-1143-x

Keywords

Search

Navigation