rAAV8-733-Mediated Gene Transfer of CHIP/Stub-1 Prevents Hippocampal Neuronal Death in Experimental Brain Ischemia

• Published in: Molecular therapy 2017-02, Vol.25 (2), p.392-400
• Main Authors: Cabral-Miranda, Felipe, Nicoloso-Simões, Elisa, Adão-Novaes, Juliana, Chiodo, Vince, Hauswirth, William W., Linden, Rafael, Chiarini, Luciana Barreto, Petrs-Silva, Hilda
• Format: Article
• Language: English
• Subjects: Agr; AKT protein; Animals; Apoptosis; brain ischemia; Brain Ischemia - genetics; Brain Ischemia - metabolism; Brain Ischemia - pathology; Cell Death - genetics; Cerebral blood flow; CHIP; Dependovirus - classification; Dependovirus - genetics; Disease Models, Animal; eIF2α; Endoplasmic reticulum; Experiments; Gene Expression; Gene therapy; Gene transfer; Gene Transfer Techniques; Genetic Therapy; Genetic Vectors - administration & dosage; Genetic Vectors - genetics; Glucose - metabolism; Hippocampus; Hypoxia; Hypoxia - metabolism; Ischemia; Kgr; Kinases; Mortality; Neurodegeneration; neuronal death; Original; Oxygen - metabolism; Phosphorylation; Protein synthesis; Proteins; Proto-Oncogene Proteins c-akt - metabolism; Pyramidal Cells - metabolism; Pyramidal Cells - pathology; rAAV; Rats; Serine; Tgr; Transduction, Genetic; ubiquitin-proteasome system; Ubiquitin-protein ligase; Ubiquitin-Protein Ligases - genetics; Ubiquitination
• ISSN: 1525-0016; 1525-0024
• DOI: 10.1016/j.ymthe.2016.11.017

Brain ischemia is a major cause of adult disability and death, and it represents a worldwide health problem with significant economic burden for modern society. The identification of the molecular pathways activated after brain ischemia, together with efficient technologies of gene delivery to the CNS, may lead to novel treatments based on gene therapy. Recombinant adeno-associated virus (rAAV) is an effective platform for gene transfer to the CNS. Here, we used a serotype 8 rAAV bearing the Y733F mutation (rAAV8-733) to overexpress co-chaperone E3 ligase CHIP (also known as Stub-1) in rat hippocampal neurons, both in an oxygen and glucose deprivation model in vitro and in a four-vessel occlusion model of ischemia in vivo. We show that CHIP overexpression prevented neuronal degeneration in both cases and led to a decrease of both eIF2α (serine 51) and AKT (serine 473) phosphorylation, as well as reduced amounts of ubiquitinated proteins following hypoxia or ischemia. These data add to current knowledge of ischemia-related signaling in the brain and suggest that gene therapy based on the role of CHIP in proteostasis may provide a new venue for brain ischemia treatment. [Display omitted] This work explored the potential use of CHIP as a gene therapy candidate. It provided the proof of principle that overexpression of CHIP promotes neuroprotection against ischemic injury in vitro and in vivo. It also explored some potential venues to explain how CHIP is working.