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Cristhian J Ildefonso, Emily E Brown, Ryo Iwata, Chulbul M Ahmed, Michael Massengill, Manas Biswal, Shannon Elizabeth Boye, William W Hauswirth, John Ash, Qiuhong Li, Alfred S Lewin; Induction of Endogenous Nrf2 Activation in the Retina Partially Protects from Oxidative Stress and Inflammation. Invest. Ophthalmol. Vis. Sci. 2016;57(12):5399.
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© ARVO (1962-2015); The Authors (2016-present)
Oxidative stress and subsequent inflammatory processes have been linked to many retinal diseases. Some important antioxidant genes are regulated by the Nrf2 transcription factor which is inhibited by Kelch-Like ECH-Associated Protein 1 (Keap-1). Herein we characterize the antioxidant and anti-inflammatory properties of an adeno-associated virus (AAV) vector delivering an Nrf2-derived peptide that binds Keap-1 and liberates endogenous Nrf2.
The Nrf2 peptide sequence was fused to that of the HIV-Tat cell-penetrating peptide (TatNrf2mer). Lentiviral-delivered TatNrf2mer was studied in cell culture. Transcript (qRT-PCR), protein levels (ELISA and immunofluoresence), and cell viability (MTT and Cell Titer assays) were quantified. Capsid modified AAV2 vectors were packaged with the TatNrf2mer fused to secretable GFP under the control of the small CBA promoter (AAV-TatNrf2mer). These vectors were evaluated in the sodium iodate model (NaIO3) of RPE oxidative injury and in the endotoxin-induced uveitis (EIU) mouse model after intravitreal injection.
TatNrf2mer expression induced antioxidant gene expression, protected cells from oxidative injury, and inhibited IL-1β secretion. In vivo, AAV-TatNrf2mer partially protected retinal function as measured by ERG responses and structure as measured by SD-OCT in the NaIO3 model. Furthermore, AAV-TatNrf2mer significantly decreased IL-1β, TNF-α, and IL-6 in the retina of NaIO3 treated mice. In the EIU mouse model, AAV-TatNrf2mer decreased the number of inflammatory cells in the vitreous body by 54%.
Intravitreal delivery of AAV-TatNrf2mer demonstrated antioxidant and anti-inflammatory effects in two models of ocular injury. This vector could be of benefit in ocular diseases in which oxidative stress and inflammation have important pathological roles.
This is an abstract that was submitted for the 2016 ARVO Annual Meeting, held in Seattle, Wash., May 1-5, 2016.
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