A targeted inhibitor of the alternative complement pathway reduces angiogenesis in a mouse model of age-related macular degeneration

Invest Ophthalmol Vis Sci. 2009 Jul;50(7):3056-64. doi: 10.1167/iovs.08-2222. Epub 2009 Mar 5.

Abstract

Purpose: Polymorphisms in factor H (fH), an inhibitor of the alternative pathway (AP) of complement activation, are associated with increased risk for age-related macular degeneration (AMD). The authors investigated the therapeutic use of a novel recombinant form of fH, CR2-fH, which is targeted to sites of complement activation, in mouse choroidal neovascularization (CNV). CR2-fH consists of the N terminus of mouse fH, which contains the AP-inhibitory domain, linked to a complement receptor 2 (CR2) targeting fragment that binds complement activation products.

Methods: Laser-induced CNV was analyzed in factor-B-deficient mice or in mice treated with CR2-fH, soluble CR2 (targeting domain), or PBS. CNV progression was analyzed by molecular, histologic, and electrophysiological readouts.

Results: Intravenously administered CR2-fH reduced CNV size, preserved retina function, and abrogated the injury-associated expression of C3 and VEGF mRNA. CR2 and PBS treatment was without effect. In therapeutically relevant paradigms involving delayed treatment after injury, CR2-fH was effective in reducing CNV and provided approximately 60% of the amount of protection of that seen in factor B-deficient mice that lacked functional AP. After intravenous injection, CR2-fH localized to sites of C3 deposition in RPE-choroid.

Conclusions: Specific inhibition of the AP reduces angiogenesis in mouse CNV. Of note, intravenous injection of C3d-targeted CR2-fH is protective even though endogenous fH is present in serum at a higher relative concentration, and serum fH contains native C3d and cell surface binding domains that target it to cell surfaces. The most common AMD-associated variant of fH resides within a native cell-binding region of fH (Tyr402His). These data may open new avenues for AMD treatment strategies.

Publication types

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

MeSH terms

  • Animals
  • Choroid / metabolism
  • Choroidal Neovascularization / drug therapy*
  • Choroidal Neovascularization / physiopathology
  • Complement Activation / drug effects
  • Complement C3 / antagonists & inhibitors
  • Complement C3 / genetics
  • Complement Factor H / antagonists & inhibitors
  • Complement Pathway, Alternative / drug effects*
  • Complement Pathway, Alternative / immunology
  • Disease Models, Animal*
  • Electroretinography
  • Female
  • Fluorescent Antibody Technique, Indirect
  • Infusions, Intravenous
  • Macular Degeneration / drug therapy*
  • Macular Degeneration / physiopathology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Mice, Knockout
  • RNA, Messenger / metabolism
  • Recombinant Fusion Proteins / administration & dosage*
  • Recombinant Fusion Proteins / pharmacokinetics
  • Retina / physiopathology
  • Retinal Pigment Epithelium / metabolism
  • Reverse Transcriptase Polymerase Chain Reaction
  • Up-Regulation
  • Vascular Endothelial Growth Factor A / antagonists & inhibitors
  • Vascular Endothelial Growth Factor A / genetics

Substances

  • CR2-Crry fusion protein, mouse
  • Complement C3
  • RNA, Messenger
  • Recombinant Fusion Proteins
  • Vascular Endothelial Growth Factor A
  • vascular endothelial growth factor A, mouse
  • Complement Factor H