Functional Studies of Dicer1 and Alu RNA in Geographic Atrophy - Adm Supplement

In this One-year Administrative Supplement to NEI R01 (PA-12-217), we will carry out additional experiments that will be crucial to defining the mechanisms of Alu RNA-induced RPE cell degeneration. In doing so, we expect that our work will lead to the optimization of neuroprotective therapeutics for the atrophic form of age-related macular degeneration (AMD). Specifically, we will determine the cellular protein-binding partners of Alu RNA by mass spectrometry, yeast three-hybrid systems, and cross-linking immunoprecipitation. In addition, we will assess whether Alu RNA itself is cytotoxic, or rather whether the conversion from Alu RNA to Alu DNA is a critical mediator of cell death. This approach will be complemented by efforts to block the RNA-DNA conversion with known retrotransposition inhibitors, which could serve as an attractive therapeutic target. Together, these approaches promise to greatly enhance the understanding of the inciting molecular events that lead to Alu RNA-induced cell death, a process that is a key determinant of AMD yet remains incompletely understood.