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Comparative Study Of Adeno-Associated Virus, Adenovirus, Baculovirus And Lentivirus Vectors For Gene Therapy Of The Eyes.

Comparative Study Of Adeno-Associated Virus, Adenovirus, Baculovirus And Lentivirus Vectors For Gene Therapy Of The Eyes.

Curr Gene Ther. 2017 Oct 03;:

Authors: Kalesnykas G, Kokki E, Alasaarela L, Lesch HP, Tuulos T, Kinnunen K, Uusitalo H, Airenne K, Ylä-Herttuala S

Background The eye possesses unique anatomical features that make it a valuable target for gene therapy applications. Objective The aim of the current study was to compare transduction efficiency, safety and biodistribution of four viral vectors following intravitreal injection. Method Adenovirus (AdV), adeno-associated virus (AAV), baculovirus (BV) and lentivirus (LV) vectors encoding green fluorescent protein (GFP) were injected bilaterally intravitreally into adult C57BL/6OlaHsd mice. Control mice received saline. Eyes and other organs were studied at multiple time points from 3 days to 6 months. Immunohistochemical stainings with retinal cell markers were performed to verify GFP-positive cells. Biodistribution inp retina and various non-target tissues was studied using a qPCR method. Inflammatory responses and toxicity were investigated from cryostat eye sections and serum samples. Results AAV-injected eyes showed GFP expression both in inner and outer retinal cells from 7 days up to 6 months. LV eyes showed long lasting transgene expression mostly in RPE cells whereas AdV transiently transduced mainly cells in the anterior chamber. In BV-injected eyes GFP positivity was very low. qPCR results showed that AdV, AAV and LV spread into the optic nerve, but were below the detection limit in other organs. The strongest immune responses were evoked by intravitreal injections of AdV and BV. The highest concentration of anti-GFP IgG was detected in the AdV-treated group, whereas the AAV group showed the lowest concentration. Neither blood chemistry screen nor the number of apoptotic cells showed any differences between the viral vector and saline injected groups. Conclusion Our findings show that intravitreal gene delivery is safe and feasible with AAV, AdV and lentivirus vectors.

PMID: 28982327 [PubMed - as supplied by publisher]