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Subconjunctival Administration of Adeno-associated Virus Vectors in Small Animal Models
Published on: March 16, 2022
Gene delivery of a viral anti-inflammatory protein to combat ocular inflammation
Cristhian J Ildefonso1, Henrique Jaime, Masmudur M Rahman
11 Department of Molecular Genetics and Microbiology, University of Florida College of Medicine , Gainesville, FL 32610.
Abstract:
Inflammation of the retina is a contributing factor in ocular diseases such as uveitis, diabetic retinopathy, and age-related macular degeneration (AMD). The M013 immunomodulatory protein from myxoma virus has been shown to interfere with the proinflammatory signaling pathways involving both the NLRP3 inflammasome and NF-κB. We have developed and characterized an adeno-associated viral (AAV) vector that delivers a secretable and cell-penetrating form of the M013 protein (TatM013). The expressed TatM013 protein was secreted and blocked the endotoxin-induced secretion of interleukin (IL)-1β in monocyte-derived cells and the reactive aldehyde-induced secretion of IL-1β in retinal pigment epithelium cells. The local anti-inflammatory effects of AAV-delivered TatM013 were evaluated in an endotoxin-induced uveitis (EIU) mouse model after intravitreal injection of mice with an AAV2-based vector carrying either TatM013 fused to a secreted green fluorescent protein (GFP) tag (sGFP-TatM013) or GFP. Expression of the sGFP-TatM013 transgene was demonstrated by fluorescence funduscopy in living mice. In EIU, the number of infiltrating cells and the concentration of IL-1β in the vitreous body were significantly lower in the eyes injected with AAV-sGFP-TatM013 compared with the eyes injected with control AAV-GFP. These results suggest that a virus-derived inhibitor of the innate immune response, when delivered via AAV, could be a generalized therapy for various inflammatory diseases of the eye.
Insights
A novel adeno-associated viral (AAV) vector delivers the M013 protein (TatM013) to reduce inflammation in the eye. This therapy significantly decreased inflammatory cells and interleukin-1β in a mouse model of uveitis.
Area of Science:
- Ophthalmology
- Immunology
- Gene Therapy
Background:
- Retinal inflammation contributes to ocular diseases like uveitis, diabetic retinopathy, and age-related macular degeneration (AMD).
- The myxoma virus M013 protein modulates NLRP3 inflammasome and NF-κB pathways, inhibiting inflammation.
- Adeno-associated virus (AAV) vectors are promising for ocular gene delivery.
Purpose of the Study:
- To develop and characterize an AAV vector for delivering a secretable and cell-penetrating form of M013 (TatM013).
- To evaluate the anti-inflammatory efficacy of AAV-delivered TatM013 in an endotoxin-induced uveitis (EIU) mouse model.
Main Methods:
- Constructed an AAV2 vector expressing a secreted, cell-penetrating M013 protein (TatM013).
- Tested TatM013's ability to block interleukin-1β (IL-1β) secretion in vitro.
- Administered AAV-sGFP-TatM013 via intravitreal injection in an EIU mouse model.
- Assessed transgene expression using fluorescence funduscopy and quantified inflammatory markers in ocular tissues.
Main Results:
- Expressed TatM013 protein was secreted and inhibited IL-1β secretion in both monocyte-derived and retinal pigment epithelium cells.
- Fluorescence funduscopy confirmed in vivo expression of sGFP-TatM013 in mice.
- Intravitreal injection of AAV-sGFP-TatM013 significantly reduced infiltrating cells and IL-1β levels in the vitreous of EIU mice compared to controls.
Conclusions:
- AAV-delivered TatM013 effectively reduces ocular inflammation in a mouse model.
- This approach demonstrates potential as a generalized gene therapy for inflammatory eye diseases.
- Virus-derived immunomodulators delivered via AAV offer a novel therapeutic strategy for ocular inflammation.
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