First Author: Z.Soheili IRAN
Co Author(s): E. Ranaei Pirmardan H. Ahmadieh S. Samiei
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In vertebrates retina, due to the lack of regeneration systems and self-renewable cells, degenerative diseases often lead to a visual impairment. One of the aims of the regenerative medicine is transformation of a cell type to another. Pax6 transcription factor has a key role in eye development and generation of the entire retinal layer. The main aim of this study was, the investigation of Pax6 overexpression, mediated by adeno-associated virus (AAV-2), in transformation of retinal differentiated cells to retinal progenitor cells in vivo.
National institute of genetic engineering and biotechnology
Recombinant AAV virus harbouring PAX6 cDNA and a reporter gene, EGFP, was produced and purified. Virus titration was performed with flow cytometry and qPCR. Induction of acute retinal ganglion cell death and generation of mouse experimental model was performed by N-methyl D-aspartic acid (NMDA) injection. All experiments including virus injection, histology and expression analyses with RT-PCR, IHC and retinal flatmount were performed.
We achieved high, acceptable transgene expression in mouse retina. We did not observe any changes in Ki67 expression following PAX6 overexpression in untreated and treated model mice. However, we showed induction of photoreceptors to express SOX2, a persistent marker for multipotential neural stem cells.
In our settings, PAX6 overexpression could not reprogram retinal cells to progenitor cells. However, we could show induction of photoreceptors to express SOX2, a universal neural stem cell marker, that confirmed high cellular plasticity in neural retinal cells. It would be a promising result for regenerative medicine approaches in eye.