Gene delivery of a viral anti-inflammatory protein to combat ocular inflammation

Hum Gene Ther. 2015 Jan;26(1):59-68. doi: 10.1089/hum.2014.089.

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.

Publication types

  • Research Support, N.I.H., Extramural
  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Cell Line, Tumor
  • Cell-Penetrating Peptides / genetics
  • Cell-Penetrating Peptides / metabolism
  • Dependovirus / genetics
  • Disease Models, Animal
  • Gene Expression
  • Gene Order
  • Gene Products, tat / genetics*
  • Gene Products, tat / metabolism
  • Gene Transfer Techniques*
  • Genes, Reporter
  • Genetic Therapy
  • Genetic Vectors / administration & dosage
  • Genetic Vectors / genetics
  • Humans
  • Interleukin-1beta / biosynthesis
  • Mice
  • Myxoma virus / genetics
  • Ophthalmoscopes
  • Recombinant Fusion Proteins*
  • Transduction, Genetic
  • Transgenes
  • Uveitis / pathology
  • Uveitis / therapy*
  • Viral Proteins / genetics*
  • Viral Proteins / metabolism
  • tat Gene Products, Human Immunodeficiency Virus

Substances

  • Cell-Penetrating Peptides
  • Gene Products, tat
  • Interleukin-1beta
  • Recombinant Fusion Proteins
  • Viral Proteins
  • tat Gene Products, Human Immunodeficiency Virus