title: An adeno-associated virus-based intracellular sensor of pathological nuclear factor-κB activation for disease-inducible gene transfer.
abstract: Stimulation 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.
authors.affiliation: Laboratory of Experimental Neurosurgery, Université Libre de Bruxelles, Brussels, Belgium. achtarto@ulb.ac.be
authors.forename: Abdelwahed
authors.initials: A
authors.lastname: Chtarto
authors.forename: Olivier
authors.initials: O
authors.lastname: Bockstael
authors.forename: Elias
authors.initials: E
authors.lastname: Gebara
authors.forename: Katia
authors.initials: K
authors.lastname: Vermoesen
authors.forename: Catherine
authors.initials: C
authors.lastname: Melas
authors.forename: Catherine
authors.initials: C
authors.lastname: Pythoud
authors.forename: Marc
authors.initials: M
authors.lastname: Levivier
authors.forename: Olivier
authors.initials: O
authors.lastname: De Witte
authors.forename: Ruth
authors.initials: R
authors.lastname: Luthi-Carter
authors.forename: Ralph
authors.initials: R
authors.lastname: Clinkers
authors.forename: Liliane
authors.initials: L
authors.lastname: Tenenbaum
mesh_terms: D000818:Animals; D016922:Cellular Senescence; D002490:Central Nervous System / Q000378:metabolism; D002540:Cerebral Cortex / Q000166:cytology; D018076:DNA, Complementary / Q000378:metabolism; D000229:Dependovirus / Q000378:metabolism*; D004195:Disease Models, Animal; D018014:Gene Transfer Techniques; D005822:Genetic Vectors / Q000378:metabolism*; D051100:Glial Cell Line-Derived Neurotrophic Factor / Q000378:metabolism; D049452:Green Fluorescent Proteins / Q000378:metabolism; D057809:HEK293 Cells; D006801:Humans; D008297:Male; D051379:Mice; D016328:NF-kappa B / Q000378:metabolism*; D009474:Neurons / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D051381:Rats; D017208:Rats, Wistar; D013226:Status Epilepticus / Q000378:metabolism; D019076:Transgenes
chemical_list: D018076:DNA, Complementary; D051100:Glial Cell Line-Derived Neurotrophic Factor; D016328:NF-kappa B; D049452:Green Fluorescent Proteins