title: Gene delivery of a viral anti-inflammatory protein to combat ocular inflammation.
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.
authors.affiliation: 1 Department of Molecular Genetics and Microbiology, University of Florida College of Medicine , Gainesville, FL 32610.
authors.forename: Cristhian J
authors.initials: CJ
authors.lastname: Ildefonso
authors.forename: Henrique
authors.initials: H
authors.lastname: Jaime
authors.forename: Masmudur M
authors.initials: MM
authors.lastname: Rahman
authors.forename: Qiuhong
authors.initials: Q
authors.lastname: Li
authors.forename: Shannon E
authors.initials: SE
authors.lastname: Boye
authors.forename: William W
authors.initials: WW
authors.lastname: Hauswirth
authors.forename: Alexandra R
authors.initials: AR
authors.lastname: Lucas
authors.forename: Grant
authors.initials: G
authors.lastname: McFadden
authors.forename: Alfred S
authors.initials: AS
authors.lastname: Lewin
mesh_terms: D000818:Animals; D045744:Cell Line, Tumor; D057846:Cell-Penetrating Peptides / Q000235:genetics / Q000378:metabolism; D000229:Dependovirus / Q000235:genetics; D004195:Disease Models, Animal; D015870:Gene Expression; D023061:Gene Order; D015696:Gene Products, tat / Q000235:genetics* / Q000378:metabolism; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000008:administration & dosage / Q000235:genetics; D006801:Humans; D053583:Interleukin-1beta / Q000096:biosynthesis; D051379:Mice; D009233:Myxoma virus / Q000235:genetics; D019721:Ophthalmoscopes; D011993:Recombinant Fusion Proteins; D014161:Transduction, Genetic; D019076:Transgenes; D014605:Uveitis / Q000473:pathology / Q000628:therapy*; D014764:Viral Proteins / Q000235:genetics* / Q000378:metabolism; D054322:tat Gene Products, Human Immunodeficiency Virus
chemical_list: D057846:Cell-Penetrating Peptides; D015696:Gene Products, tat; D053583:Interleukin-1beta; D011993:Recombinant Fusion Proteins; D014764:Viral Proteins; D054322:tat Gene Products, Human Immunodeficiency Virus