Age-related macular degeneration (AMD)

Age-related macular degeneration (AMD) is a medical condition usually affecting older adults and resulting in a loss of vision in the macula, the centre of the visual field. A major part of the dry form of the disease (85-90% of cases) is atrophy of the retinal pigment epithelium, resulting in the detachment of the retina and loss of photoreceptors (Bhutto and Lutty, 2012). Apart from age, cigarette smoke is the main risk factor for AMD and increases the risk of developing the disease by two to three times (Smith et al., 2001; Tomany et al., 2004).

The primary aim of the project is to improve our understanding of the tobacco smoke-induced molecular mechanisms underlying RPE destruction in the context of AMD. Therefore, we want to identify up-regulated/down-regulated proteins in cultured human ARPE-19 cells that were treated with cigarette smoke extract for defined time points.

We developed a highly reproducible cell fractionation workflow, allowing for the mass spectrometric identification and quantification of secreted proteins, as well as the three cell fractions cell-surface, nucleus and cytoplasm. More than 3000 proteins can be identified in the different fractions and their respective abundances can be compared between cigarette smoke-treated and untreated cells by label-free quantitative LC-MS/MS.