Periodic Reporting for period 1 - EaSYFUN (The role of eEF1A in synapse function, maintenance, and synucleinopathy)
Okres sprawozdawczy: 2022-06-01 do 2024-11-30
EaSYFUN explores early brain alterations in neurodegenerative diseases induced by synucleinopathy such as Parkinson's disease. These diseases induce progressive loss of motor control, tremor and rigidity as well as senile dementia. Synucleinopathies are associated with the abnormal accumulation of a protein called alpha-synuclein. This accumulation in the form of aggregates generates a loss of cells (neurons) and a loss of connections between neurons (synapses), but scientists still don’t fully understand the mechanisms driving this slow degeneration.
The EaSYFUN project focuses on a protein called eukaryotic elongation factor 1 alpha (eEF1A), which is known for helping cells make other proteins by bringing the building blocks of the protein molecules to the site of synthesis. However, early findings suggest that eEF1A may play other roles in the brain, including in the response to misfolded proteins like alpha-synuclein. In mouse model of synucleinopathy, the host laboratory previously observed that eEF1A levels drop significantly when the first synapses display molecular changes in the disease progress (before any symptoms develop). The eEF1A is reduced specifically at synapses — the connections between neurons — suggesting that eEF1A loss might contribute to the development of synucleinopathies. EaSYFUN ambitions to clarify the involvement of eEF1A in normal function of synapse and how this function is then affected in pathology of synucleinopathy. By exploration of individual eEF1A possible functions in the cell, its involvement will be either eliminated or confirmed to participate to the disease triggering or progression.
EaSYFUN includes several work packages (WPs) to investigate this hypothesis further.
The EaSYFUN project focuses on a protein called eukaryotic elongation factor 1 alpha (eEF1A), which is known for helping cells make other proteins by bringing the building blocks of the protein molecules to the site of synthesis. However, early findings suggest that eEF1A may play other roles in the brain, including in the response to misfolded proteins like alpha-synuclein. In mouse model of synucleinopathy, the host laboratory previously observed that eEF1A levels drop significantly when the first synapses display molecular changes in the disease progress (before any symptoms develop). The eEF1A is reduced specifically at synapses — the connections between neurons — suggesting that eEF1A loss might contribute to the development of synucleinopathies. EaSYFUN ambitions to clarify the involvement of eEF1A in normal function of synapse and how this function is then affected in pathology of synucleinopathy. By exploration of individual eEF1A possible functions in the cell, its involvement will be either eliminated or confirmed to participate to the disease triggering or progression.
EaSYFUN includes several work packages (WPs) to investigate this hypothesis further.
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.
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.
The project aims to lay groundwork for future therapeutic options by better understanding these proteins and their roles in early synucleinopathy stages, potentially offering new ways to slow or prevent disease progression.