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Christina A. Kramann, Nils Boehm, Johannes Hummel, Markus Schlich, Katrin Lorenz, Bernhard M. Stoffelns, Norbert Pfeiffer, Franz H. Grus; Comparative Proteomic Analysis of Human Vitreous in Patients with Various Vitreoretinal Diseases by MALDI-OrbitrapTM XL. Invest. Ophthalmol. Vis. Sci. 2011;52(14):539.
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© ARVO (1962-2015); The Authors (2016-present)
To develop a proteome analysis of human vitreous samples in patients with various vitreoretinal diseases to provide insights into factors and mechanisms responsible for these diseases.
Vitreous samples were collected from 49 patients during pars plana vitrectomy of patients with macular pucker (n=14), idiopathic macular hole (n=13), proliferative diabetic retinopathy (n=8), rhegmatogenous (n=4) and tractional (n=4) retinal detachment or vitreous hemorrhage associated with age-related macular degeneration (n=3), with retinal vein occlusion (n=2) or with choroidal melanoma (n=1). For the proteomic mapping, 10 pooled vitreous samples were chosen and gel lanes of one- and two-dimensional gel electrophoresis were sectioned into slices. Each slice was subjected to enzymatic hydrolysis. The resultant mixture of peptide fragments was analyzed by matrix-assisted laser desorption time-of-flight mass spectrometry (MALDI-OrbitrapTM XL). For validation of biomarkers we used an antibody microarray approach.
Complex patterns of vitreous proteins and peptides could be detected in all groups, overall 116 proteins could be identified. Compared to the other groups the relative protein expression in the proliferative diabetic group revealed mainly an up to five-fold up- or downregulation. Compared to the macular hole group a 58-fold increase of α-1-acid Glycoprotein 1 (A1AG1) and a 25-fold decrease of α -1- Mikroglobulin (AMBP) could be detected in the proliferative diabetic group (α<0.05). In the group with tractional retinal detachment, there was a 74-fold increase in relative expression of Serpin B3 and a 13-fold increase of Serpin B4 compared to the proliferative diabetic group (α<0.05). In the macular pucker group there was a 51-fold decrease of α-1-acid Glycoprotein 1 (A1AG1) and in the macular hole group a 58-fold decrease of α-1-acid Glycoprotein 1 (A1AG1) compared to proliferative diabetic group (α<0.05).
Each vitreoretinal disease has a unique set of proteins which can be interpreted based on the pathology of retinopathy. Our study provides an analysis of the human vitreous proteome and reveals protein alterations in the group of proliferative diabetic retinopathy compared to other vitreoretinal diseases.
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