Neurotoxic · An Ultra-sensitive Assay to Measure Oligomer Induced Toxicity in Human Cerebrospinal Fluid
Horizon 2020 — Marie Skłodowska-Curie Actions
- Duration
- 2016-08-01 → 2018-07-31
- EU contribution
- €195,455
- Participants
- 1
- Scheme
- MSCA-IF-EF-ST
Lines connect the coordinator with its partners.
Results in brief
An Ultra-sensitive Assay to Measure Oligomer Induced Toxicity in Human Cerebrospinal Fluid
There are an estimated 50 million people worldwide living with dementia and the total estimated worldwide cost of dementia is $1 trillion. Alzheimer’s disease is most common cause of dementia which accounts for an estimated approximately 60% to 80% percent of cases. The conversion of peptides or proteins from their soluble native states into insoluble amyloid deposits has been described as a hallmark of many neurodegenerative disorders including Alzheimer’s disease (AD). Misfolded oligomeric intermediates, rather than matured fibrils, formed during the aggregation process have been identified as the primary pathogenic agents. It has been shown that targeting these oligomeric species with antibodies can reduce pathology in both mouse models and the human brain. This highly heterogeneous conformers of protein aggregation are transient species which only constitute a very small fraction as compared to the amyloid and the non-aggregated native form of the specific protein or peptide. Therefore, despite the importance of oligomers as the key cause of pathogenesis of many neurodegenerative diseases, it has been extremely challenging to characterize experimentally the functions and structures of these small soluble aggregates both within aggregation mixture at physiological condition (in the range of pM concentration) as well as in human bio-markers such as Cerebrospinal fluid. Moreover, due to the complex mechanism of their formation and their heterogeneous and dynamical nature, traditional bulk methods of structural biology cannot easily be applied. Therefore, it is important to develop methods that can detect and quantify the toxic form of aggregate present in the aggregation mixture as well as in complex biofluids. Such a method would then allow to study molecules that bind these toxic forms and delineate effective therapeutic strategies for protein aggregation disorders.
Data: CORDIS, © European Union
Project objective
The study of protein aggregates and how they damage neuronal cells is important in order to understand the initiation and progression of several neurodegenerative diseases - including Alzheimer’s disease (AD), Parkinson’s disease (PD). The aggregation from the native monomeric proteins to beta sheet containing amyloid structures involves the formation of different species - misfolded proteins, small soluble oligomers and finally formation of fibrils. It is believed that small oligomers are the most cytotoxic species which play a major role in neuronal loss and cell death. Although, the exact mechanism and extent of amyloid oligomer’s cytotoxicity is still unknown, one of the most consistent pathologies in neurodegenerative disease is unregulated influx of Ca2+ into the cell. Individual oligomers directly disrupt cell membranes through non-specific binding at picomolar concentrations leading to the formation of ion channels, which allow Ca2+ influx and can lead to cell death. Based on this observation, we will develop a single liposome assay to quantify the oligomer induced toxicity from oligomers present in human cerebrospinal fluid (CSF) from healthy controls and patients with AD and PD. Using a Ca2+ sensitive fluorogenic sensor, we will monitor the oligomer induced Ca2+ influx inside the liposome in real time. We will measure and compare the toxicity of synthetic oligomers of amyloid beta, alpha-synuclein, tau and PrP oligomers as a function of oligomer concentration and liposome membrane composition. We will then quantify the effect of various antibodies commonly used for treatment of AD and PD on the toxicity of synthetic oligomers and those in CSF. This novel ultra-sensitive state-of-the-art single molecule technique will provide new insights into the mechanism of oligomer induced toxicity, allowing us to identify the most toxic oligomers and the most suitable antibodies to prevent oligomer induced damage as well as having potential for early disease diagnosis.
Original text from CORDIS.
Participants
- THE CHANCELLOR MASTERS AND SCHOLARS OF THE UNIVERSITY OF CAMBRIDGE · CAMBRIDGECoordinatorUnited Kingdom
Links
- View on CORDIS
- DOI: 10.3030/701013
- https://ec.europa.eu/programmes/horizon2020/en/h2020-section/marie-sklodowska-curie-actions
Data: CORDIS, © European Union