Preparation of amyloid-beta aggregate species from synthetic and patient-derived material to define disease-causing mechanisms
Species of misfolded and aggregated proteins are pathological hallmarks of several devastating neurodegenerative disorders, such as Alzheimer's, Parkinson's or Huntington's disease. There is a large body of evidence that misfolded protein aggregates are causally related to neurodegeneration as initiators or drivers of pathological cascades. In Alzheimer's disease (AD), the amyloid-beta peptide formed by proteolytic cleavage of the amyloid precursor protein accumulates in affected brain tissue. The exact course of amyloid pathology in AD, however, remains to be elucidated. So far a variety of different brain-derived and synthetic amyloid-beta aggregate species from dimers, small and large oligomers to protofibrillar and fibrillar aggregates have been reported. This variety poses a methodical conundrum. Which type of amyloid-beta aggregate species is indeed relevant to disease and how does it mediate dysfunction and neurodegeneration? While previously the mature, end stage amyloidbeta fibrils were seen as responsible for neurodegeneration, more recently, evidence has accumulated that small, soluble oligomers are causative to disease. Many attempts have been made to isolate disease-relevant amyloid-beta oligomers from transgenic mouse and Alzheimer's patient brains. However, their very low abundance in brain precludes systematic biochemical and structural investigation. Here, we propose to develop a set of methods for the amplification of amyloid-beta aggregate species from patient brain tissue and for generating standardized fully synthetic aggregate species. We will compare and characterize these species with biochemical and biophysical methods, and determine disease-relevance and competence to seed aggregates in cell-free and in vivo models. Also, we will identify and validate small molecule modulators of the amyloid formation process to study the mechanisms of aggregation and to facilitate the exploration of new therapeutic strategies.
Imaging techniques, Pharmacology, Molecular and (epi)genetic and "omics" approaches, Amyloid-beta, protein aggregation
2013 - 2016
Erich Wanker (Coordinator)
Bart De Strooper