Detection of high risk drusen

Abstract

In some aspects, methods of detecting complement activation in vivo, e.g., in an eye, are provided. In some aspects, methods of detecting high risk drusen are provided. In some aspects, presence of one or more high risk drusen in an eye indicates increased likelihood of developing AMD, GA, or advanced AMD or increased likelihood of rapid progression of AMD. In some embodiments, methods comprise detecting drusen containing or in close proximity to complement activation. In some embodiments methods comprise detecting one or more drusen having inflamed endothelium underlying or in close proximity thereto. In some embodiments methods comprise detecting eye-derived extracellular microvesicles, e.g., exosomes, in a body fluid. In some embodiments methods comprise detecting Th17 cells or a Th17 biomarker in a body fluid. In some embodiments the Th17 biomarker is a cytokine. Methods may be applied individually or in any combination. In some embodiments any of the methods further comprises treating a subject at risk of developing AMD, GA, or advanced AMD or at increased likelihood of rapid progression of AMD with a complement inhibitor. In some aspects, agents useful for performing one or more of the methods are described.

1 . A method of detecting complement activation in a subject comprising: (a) providing a subject to whom a substrate for a complement activation pathway protease has been administered; and (b) detecting cleavage of the substrate, thereby detecting complement activation in the subject. 
     
     
         2 . (canceled) 
     
     
         3 . The method of  claim 1 , wherein step (a) comprises administering the substrate to the subject. 
     
     
         4 . The method of  claim 1 , wherein the substrate comprises a peptide substrate for a C3 convertase. 
     
     
         5 . (canceled) 
     
     
         6 . The method of  claim 1 , wherein the substrate produces a detectable signal upon cleavage; and step (b) comprises detecting the signal. 
     
     
         7 . The method of  claim 1 , wherein cleavage of the substrate produces a fluorescent, visible, magnetic, or infrared signal. 
     
     
         8 . The method of  claim 1 , wherein the substrate comprises a peptide substrate for a C3 convertase linked to a first moiety and a second moiety, wherein cleavage of the substrate alters the distance between the first and second moieties, thereby resulting in a detectable signal. 
     
     
         9 . (canceled) 
     
     
         10 . The method of  claim 1 , wherein the substrate comprises a peptide substrate for a C3 convertase linked to a first fluorophore and a second fluorophore at a characteristic distance wherein the fluorescences of the first and second fluorophores are mutually substantially quenched. 
     
     
         11 . The method of  claim 1 , wherein the substrate comprises a peptide substrate for a C3 convertase linked to fluorophore and a quencher at a characteristic distance wherein the fluorescence of the fluorophore is quenched by the quencher. 
     
     
         12 . (canceled) 
     
     
         13 . The method of  claim 1 , wherein the subject is at risk of or suffering from age-related macular degeneration (AMD), glaucoma, or uveitis. 
     
     
         14 . (canceled) 
     
     
         15 . (canceled) 
     
     
         16 . The method of  claim 1 , wherein detecting is performed using a medical imaging device. 
     
     
         17 . The method of  claim 1 , wherein detecting is performed using an ophthalmoscope. 
     
     
         18 . The method of  claim 1 , comprising detecting one or more drusen in an eye of the subject and detecting complement activation in or in close proximity to one or more of said drusen. 
     
     
         19 .- 37 . (canceled) 
     
     
         38 . A method of detecting a high risk druse in an eye of a subject, the method comprising detecting complement activation in or in close proximity to a druse in an eye of a subject or detecting inflamed endothelium underlying or in close proximity to a druse in an eye of a subject. 
     
     
         39 . The method of  claim 38 , comprising detecting complement activation in or in close proximity to the druse. 
     
     
         40 . The method of  claim 38 , comprising detecting cleavage of a substrate for a complement activation pathway protease in or in close proximity to the druse. 
     
     
         41 .- 52 . (canceled) 
     
     
         53 . A method of identifying an eye at increased risk of developing advanced AMD, the method comprising detecting complement activation in or in close proximity to one or more drusen in an eye of a subject in vivo or detecting inflamed endothelium underlying or in close proximity to one or more drusen in an eye of a subject in vivo. 
     
     
         54 .- 126 . (canceled) 
     
     
         127 . A method of identifying a subject at increased risk of development or progression of AMD, the method comprising: detecting an increased level of eye-derived extracellular vesicles (EVs) in a body fluid of the subject, wherein an increased level of said EVs is indicative that the subject is at increased risk of development or progression of AMD. 
     
     
         128 . The method of  claim 127 , wherein at least some of the eye-derived EVs originate from retinal cells or retinal pigment epithelial (RPE) cells. 
     
     
         129 . (canceled) 
     
     
         130 . The method of  claim 127 , wherein the EVs comprise at least one cell-type specific marker for RPE cells or retinal cells. 
     
     
         131 . (canceled) 
     
     
         132 . The method of  claim 127 , wherein the EVs comprise at least one cell-type specific marker for RPE cells or retinal cells, wherein the marker is an eye-specific opsin. 
     
     
         133 .- 136 . (canceled) 
     
     
         137 . The method of  claim 127 , wherein the EVs comprise exosomes. 
     
     
         138 .- 141 . (canceled) 
     
     
         142 . The method of  claim 127 , further comprising administering a complement inhibitor or anti-Th17 agent to the subject based at least in part on determining that the subject is at increased risk of developing or progression of AMD. 
     
     
         143 .- 146 . (canceled)