Key Parts of the Project
1. WP1 focuses on understanding where and when eEF1A proteins are reduced in mouse models of synucleinopathy. Using multiple antibodies for eEF1A detection (differing between EEF1A1 and EEF1A2), researchers tracked the protein presence at several ages before and after disease onset. Disease onset was defined by the first quantitative detection of synapse loss (18 weeks in the model). They saw a different behavior between the 2 eEF1A proteins and a notable decrease at synapses in one type only around the time of disease onset. For the confirmation of the protein variant involved, genetic fluorescent labeling of the cellular protein was developed which will enable the spatial localization to specific site of the synapse. To study human cells, the Fellow has also set up stem cell laboratory to enable to create lab-grown human neurons and trained staff to work with these models.
2. WP2 aims to investigate how eEF1A loss affects brain cell functions, especially how it might impact synapse health. The main function of the eEF1A protein being to assist the delivery of amino-acids, the building blocks of proteins, to ribosomes, the site of protein synthesis. EaSYFUN thus aimed at testing the ability of brain cells to produce new proteins while eEF1A level is reduced. We found that the protein production remains unchanged in healthy and diseased brain tissue even at synapses. Synaptic vesicles are essential for neurotransmission by supporting neurotransmitter accumulation and regulated secretion. The team suspected that eEF1A loss may affect neurotransmitter secretion. EaSYFUN tested this hypothesis by comparing the disease mouse model with healthy animals.
Another significant focus of WP2 is understanding how eEF1A might help cells to manage the elimination of misfolded alpha-synuclein. Misfolded proteins can clump together, damaging cells, and should be removed from the cell by the regulatory mechanisms of the cell. EaSYFUN incorporates a fluorescent reporter of alpha-synuclein which allows to monitor the level of the protein in the cell under a microscope. EaSYFUN intends to test the levels of alpha-synuclein in the disease mouse model compared with healthy animals. Moreover, the human stem cell derived neurons becomes the perfect candidate for this experiment, and neurons from disease donors as well as from genetically corrected lines will be compared.
3. WP3 aims at investigating the effects of a modulation of the eEF1A proteins independent of a model of synucleinopathy. We tested different drugs to alter eEF1A activity, as well as genetic manipulation of EEF1A protein expression (removal, rescue and overexpression) on several cell functions.
