Oligomerization of alpha-synuclein is hypothesized to be the pathogenic mechanism of protein aggregate formation and cell death in neurons of PD patients. Dr. Outeiro proposes to develop an assay to screen for specific genes that affect alpha-synuclein oligomerization. Dr. Outeiro and colleagues developed a biofluorescence measurement of alpha-synuclein oligomerization in mammalian cells. Using the fluorescence endpoint, he will screen the human genome using siRNA technology to uncover genes that alter oligomerization. Positive hits will potentially lead to the identification of new targets for therapeutics attempting to block alpha-synuclein aggregation.
The formation of large alpha-synuclein protein inclusions in the brain is the pathological hallmark of Parkinson’s disease but whether those inclusions are the actual culprits remains unknown. We now have reason to believe that smaller forms of alpha-synuclein with altered structure, known as oligomers, may be more toxic for cells.
Dr. Outeiro and colleagues set out to establish a system in which to visualize these smaller forms, which had not been observed in living cells. They carried out a pilot RNAi screen and identification of genes which interfere with the formation of those forms of alpha-synuclein, either promoting or inhibiting their formation. Through his screen, Dr. Outeiro identified initial hits that may represent novel therapeutic targets for PD. He received supplemental funding to validate these initial positive hits.