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Nour Maya Haddad, Jennifer Sun, Michael Molla, Paul Arrigg, Sabera Shah, Deborah Schlossman, Timothy Murtha, Edward Feener, Lloyd Aiello; Correlation of the Aqueous and Vitreous Proteomes in Diabetic Eye Disease. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1128. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To compare proteomes between aqueous fluid (AF) and vitreous fluid (VF) obtained concurrently from the same eye of patients undergoing ocular surgery and to assess differences between diabetic retinopathy (DR) severity levels.
AF and VF were obtained from 6 eyes (2 each NPDR, active PDR and quiescent PDR) undergoing combined cataract surgery and pars plana vitrectomy. AF (0.05-0.10cc) was obtained via anterior chamber paracentesis. Cataract surgery was performed after which ~1cc undiluted VF was obtained through the vitrector before initiating infusion. All samples were stored at -80°F until batch processed. Proteomic analysis used 30μL of undiluted sample. Proteins were analyzed by tandem mass spectrometry and protein levels compared using peptide/spectral matches assigned using X!Tandem.
228 unique proteins were identified overall in AF and VF with 110 (48%) found in both AF&VF, 16 (7%) in AF only, and 102 (45%) in VF only. The percentage of proteins found in both AF&VF for NPDR, PDR and QPDR groups was 50% (88), 48 % (80) and 58% (67), respectively. VF proteins were present only in VF in 37% (64) of NPDR, 47% (79) of PDR, and 32% (37) of QPDR. Overall, 52% (110) of VF detected proteins were also detectable in the AF. When proteins were present in both AF&VF, peptide counts between VF and AF for an individual protein were highly correlated (r=0.76-0.89) and correlations did not differ substantially between DR groups (r = 0.83-0.84). Peptide counts were higher in VF than AF in 87.3% of proteins for the combined group, 92% in NPDR, 92% in PDR and 76% in QPDR. On average, peptide counts for VF predominant proteins were 5 fold higher in VF than AF for any specific protein. A partial survey of specific proteins known to be predominantly vitreal had VF/AF ratios >1.0 with peptide numbers changing by group as expected for changes in disease severity (eg. carbonic anhydrase 1, C1 inhibitor).
This study reveals that approximately 50% of the proteins found in vitreous can also be detected in aqueous. There was also a high correlation between proteins found in both the AF and VF, and AF findings correlated with known VF changes for at least some specific proteins. These data suggest that AF sampling might allow proteomic assessment reflecting the vitreome in the diabetic eye and help evaluate specific vitreous proteins as biomarkers of DR progression or treatment response.
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