An adeno-associated virus-based intracellular sensor of pathological nuclear factor-κB activation for disease-inducible gene transfer..pdf (7.69 MB)
Download fileAn adeno-associated virus-based intracellular sensor of pathological nuclear factor-κB activation for disease-inducible gene transfer.
journal contribution
posted on 2020-06-25, 07:19 authored by A Chtarto, O Bockstael, E Gebara, K Vermoesen, C Melas, C Pythoud, M Levivier, O De Witte, R Luthi-Carter, R Clinkers, L TenenbaumStimulation of resident cells by NF-κB activating cytokines is a central element of inflammatory and degenerative disorders of the central nervous system (CNS). This disease-mediated NF-κB activation could be used to drive transgene expression selectively in affected cells, using adeno-associated virus (AAV)-mediated gene transfer. We have constructed a series of AAV vectors expressing GFP under the control of different promoters including NF-κB -responsive elements. As an initial screen, the vectors were tested in vitro in HEK-293T cells treated with TNF-α. The best profile of GFP induction was obtained with a promoter containing two blocks of four NF-κB -responsive sequences from the human JCV neurotropic polyoma virus promoter, fused to a new tight minimal CMV promoter, optimally distant from each other. A therapeutical gene, glial cell line-derived neurotrophic factor (GDNF) cDNA under the control of serotype 1-encapsidated NF-κB -responsive AAV vector (AAV-NF) was protective in senescent cultures of mouse cortical neurons. AAV-NF was then evaluated in vivo in the kainic acid (KA)-induced status epilepticus rat model for temporal lobe epilepsy, a major neurological disorder with a central pathophysiological role for NF-κB activation. We demonstrate that AAV-NF, injected in the hippocampus, responded to disease induction by mediating GFP expression, preferentially in CA1 and CA3 neurons and astrocytes, specifically in regions where inflammatory markers were also induced. Altogether, these data demonstrate the feasibility to use disease-activated transcription factor-responsive elements in order to drive transgene expression specifically in affected cells in inflammatory CNS disorders using AAV-mediated gene transfer.
Funding
This work was supported by FNRS-Te ́le ́vie: grant number: 7.4.593.08 F. ‘‘Cre ́dit aux chercheurs’’ from the Belgian National Research Foundation: grantnumber: 1.5.093.08, ‘‘Fonds National de la Recherche Scientifique Me ́dicale’’ grant number 7.4.536.09 F. ‘‘Re ́gion Bruxelles-Capitale’’ (SOIB) grant REGAVEC 2007–10, Swiss National Research Foundation (FNS) grant number: FN31003A-127177, Agency for Innovation by Science and Technology (IWT Vlaanderen), andResearch Foundation Flanders (FWO Vlaanderen).
History
Citation
PLoS One, 2013, 8 (1), e53156Author affiliation
Neuroscience, Psychology and BehaviourVersion
- VoR (Version of Record)
Published in
PLoS OneVolume
8Issue
1Publisher
Public Library of Scienceeissn
1932-6203Acceptance date
2012-11-26Copyright date
2013Available date
2013-01-03Publisher DOI
Language
engAdministrator link
Publisher version
http://www.ncbi.nlm.nih.gov/pubmed/23301037Usage metrics
Categories
Keywords
AnimalsCell AgingCentral Nervous SystemCerebral CortexDNA, ComplementaryDependovirusDisease Models, AnimalGene Transfer TechniquesGenetic VectorsGlial Cell Line-Derived Neurotrophic FactorGreen Fluorescent ProteinsHEK293 CellsHumansMaleMiceNF-kappa BNeuronsPromoter Regions, GeneticRatsRats, WistarStatus EpilepticusTransgenes