Growth-compromised HSV-2 vector Delta RR protects from N-methyl-D-aspartate-induced neuronal degeneration through redundant activation of the MEK/ERK and PI3-K/Akt survival pathways, either one of which overrides apoptotic cascades
| Autor: | Juan Liu, Jennifer M. Laing, Paul J. Yarowsky, Samantha Q. Wales, Erin K. Golembewski, M. Samir Jafri, Laure Aurelian |
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| Rok vydání: | 2007 |
| Předmět: |
MAPK/ERK pathway
Programmed cell death N-Methylaspartate Cell Survival Herpesvirus 2 Human Genetic Vectors Immunoblotting Fluorescent Antibody Technique Apoptosis Biology Protein Serine-Threonine Kinases Neuroprotection Rats Sprague-Dawley Cellular and Molecular Neuroscience chemistry.chemical_compound Phosphatidylinositol 3-Kinases Chlorocebus aethiops Ribonucleotide Reductases medicine Excitatory Amino Acid Agonists In Situ Nick-End Labeling Animals LY294002 Protein kinase B Vero Cells Neurons MEK inhibitor Neurodegeneration Genetic Therapy medicine.disease Molecular biology Corpus Striatum Cell biology Rats chemistry Nerve Degeneration Mitogen-Activated Protein Kinases |
| Zdroj: | Journal of neuroscience research. 86(2) |
| ISSN: | 1097-4547 |
| Popis: | We have previously shown that intrastriatal injection of Delta RR, the growth-compromised herpes simplex virus type 2 (HSV-2) vector for the antiapoptotic protein ICP10PK, prevents apoptosis caused by the excitotoxin N-methyl-D-aspartate (NMDA) in a mouse model of glutamatergic neuronal cell death (Golembewski et al. [2007] Exp. Neurol. 203:381-393). Because apoptosis regulation is stimulus and cell type specific, our studies were designed to examine the mechanism of Delta RR-mediated neuroprotection in striatal neurons. Organotypic striatal cultures (OSC) that retain much of the synaptic circuitry of the intact striatum were infected with Delta RR or a growth-compromised HSV-2 vector that lacks ICP10PK (Delta PK) and examined for neuroprotection-associated signaling. The mutated ICP10 proteins (p175 and p95) were expressed in 70-80% of neurons from Delta RR- and Delta PK-infected cultures, respectively, as determined by double-immunofluorescent staining with antibodies to ICP10 and NeuN or GAD65. Delta RR- but not Delta PK-treated OSC were protected from NMDA-induced apoptosis, as verified by ethidium homodimer staining, TUNEL, caspase-3 activation, and poly(AD-ribose) polymerase (PARP) cleavage. Neuroprotection was through ICP10PK-mediated activation of the survival pathways MEK/ERK and PI3-K/Akt, up-regulation of the antiapoptotic proteins Bag-1 and Bcl-2, and phosphorylation (inactivation) of the proapoptotic protein Bad. It was blocked by the MEK inhibitor U0126 or the PI3-K inhibitor LY294002, suggesting that either pathway can prevent NMDA-induced apoptosis. The data indicate that Delta RR-delivered ICP10PK stimulates redundant survival pathways that override proapoptotic cascades. Delta RR is a promising gene therapy platform against glutamatergic cell death. |
| Databáze: | OpenAIRE |
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