Identifying and deciphering the mechanism of the cellular machinery responsible for disaggregation of intracellular prion-like Tau aggregates

Objectif

Tauopathies are a group of neurodegenerative diseases, the most common of them being Alzheimer’s disease, which are associated with the aggregation of protein Tau in the brain. Despite major research efforts world-wide, the mechanism of Tau aggregation and its pathogenesis giving rise to such neurodegenerative diseases is not yet understood. Therapeutics to cure these tauopathies, including Alzheimer’s disease, are urgently needed. We aim to identify the conditions, the cellular machinery responsible and the mechanism of disaggregation of intracellular prion-like Tau aggregates in HEK293 cell lines, with one condition already been identified (starvation). For identifying the conditions of disaggregation, high resolution confocal microscopy will be used for visualizing intracellular Tau aggregates tagged with yellow fluorescence protein (YFP). For deciphering the cellular components responsible for disaggregation, the proteins associated with the aggregates will be identified using quantitative mass spectrometry from SILAC (stable isotope labeling with amino acids in cell culture) labeled cells. Moreover, candidate factors will be downregulated by siRNA to establish their involvement in aggregate disassembly. Such components individually and in combination will then be analyzed for their disaggregation activity in vitro using filter trap assay for the quantification of disaggregation. Cryo-electron tomography of Tau aggregates under various conditions (e.g. during disaggregation) will be carried out in collaboration with Prof. Wolfgang Baumeister (MPIB). Besides contributing to the benefit of humankind, the proposed research work on neurodegenerative diseases will allow me to grow as a researcher and will help me in realizing my true potential as a scientist. The MPIB and the Hartl lab in particular will play an important role in this regard due the availability of state-of-the-art research facilities and an intellectually stimulating environment.