WP1 Preparation of tau filament models relevant to individual tauopathies in vitro, ex vivo and in vivo
The specific aims of WP1 are:
- To prepare source material - pure and characterized monomeric Tau, native and/or phosphorylated, isotopically labelled
- To prepare assembled Tau prepared from isotopically labelled recombinant material
- To prepare cell lines and neuronal primary cell cultures harboring isotopically labeled Tau monomers and oligomers for in-cell NMR experiments
- To isolate assembled Tau from the brain tissue of transgenic animals
- To prepare interaction partners of Tau and disordered proteins (e.g., Map2c) for use as controls
WP2 Investigation of key structural changes in soluble Tau leading to pathologic aggregation
The specific aims of WP2 are:
- To dissect the specific structural profile of Tau variants in solution state
- To compare structural behavior of Tau variants in cells with respect to (in vitro) buffer conditions
- To determine phosphorylation patterns of Tau variants and its influence on tau structure
- To determine tau binding epitopes with respect to its binding partners (tubulin, 14-3-3)
- To elucidate monomeric-oligomeric state of tau by molecular dynamics of selected tau fragments
WP3 Comprehensive characterization of Tau filament structure
The specific aims of WP3 are:
- To determine the high-resolution structures of various tau filaments by cryo-EM
- To characterize interaction of the tau filament with binding partners and antibodies by EM
- To determine the unique molecular structure of pathognomonic fibrillary tau variants by solid-state NMR.
- To evaluate interactions of assembled tau with various partners, map the binding site and secondary structure
- changes upon complex formation
- To create the structural models of tau in various solid forms (oligomers, fibrils, co-aggregates)
WP4 Translation of tau structural changes to cellular signalling networks and interneuronal tau propagation
The specific aims of WP4 are:
- To develop novel systems for simultaneous monitoring of interneuronal tau spreading and its effect on cellular physiology, to test the potency of different tau species, and to explore the effects of therapeutic interventions
- To characterise the cellular mechanisms involved in this pathological process
- To predict novel therapeutic targets and diagnostic biomarkers using experimental data from proprietary animal models of tauopathy
- To identify in silico validated targets and pathway models in which different tau species and 14-3-3
- proteins are involved using integrative computational biology
- To validate interaction partners and signalling pathways predicted in silico using experimental cell systems
- To analyse large miRNA datasets from animal models by state of the art bioinformatic tools
WP5 Integration and exploitation
The specific aims of WP5 are:
- To analyse proposed mode of action of current anti Tau vaccines with respect to the results obtained in the project
- To exploit acquired knowledge to formulate new hypotheses in therapy
WP6 Management and dissemination
This project has received funding from the European Union’s Horizon 2020 research and innovation programme under the Marie Skłodowska-Curie grant agreement no. 873127 – InterTAU.