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Tuning Tubulin Dynamics and Interactions to Face Neurotoxicity: a Multidisciplinary Approach for Training and Research

Periodic Reporting for period 1 - TubInTrain (Tuning Tubulin Dynamics and Interactions to Face Neurotoxicity: a Multidisciplinary Approach for Training and Research)

Período documentado: 2019-10-01 hasta 2021-09-30

The improvements in life expectancy in the Western societies have turned neurodegenerative diseases and cancer into the most common health problems of the European Union. It is therefore of outmost importance to find and optimize tools that can contribute to the full comprehension of the molecular mechanisms involved in the progression of these pathologies.
Changes in the behavior and organization of microtubules (MTs) are emerging as relevant events associated with many neurodegenerative diseases.
Deciphering the very basic pathological mechanisms of neuronal damage linked to tubulin/microtubule perturbation is a challenge for facing neurodegeneration.

The aim of TubInTrain is to supply a transdisciplinary view of the tubulin-targeting strategy for neuron functionality (Fig.2). This is mandatory for a better comprehension of MTs involvement in the pathogenic mechanisms of neurodegeneration. We are confident that TubInTrain will also contribute to the design of mechanism-based therapeutic interventions, thus making this project in line with the aim of Horizon 2020 programme [Challenge “Health, demographic change and wellbeing”]. In particular, we will investigate different strategies for tuning tubulin dynamics and interactions aiming to:
i) model and evaluate the impact of tubulin interacting compounds on tubulin/MTs at atomic level in cell free system,
ii) model and evaluate the impact of two tubulin interacting proteins (Tau and alpha-Synuclein) on tubulin/MTs at atomic level in cell free system,
iii) determine the effects of the above compounds on MT dynamics, protein-MT interaction, protein aggregation and metabolism in peripheral neurons and in cellular models of neurodegeneration.

The research activities are articulated into three Work Packages (WP1-WP2-WP3 - Fig.3)
The work performed by the network could briefly summarized as subsequently reported:

1) The computational studies allow to a) define a fruitful strategy to virtually screen large library of compounds and to design new promising compound able to interact with tubulin/MTs and b) face the behaviour of alpha-synuclein mutants in comparison to native alpha-synuclein

2) The research groups involved in the chemical synthesis prepare some de-novo designed compounds and some putative α-Synuclein MTs binding peptides

3) X-Ray crystallography demonstrate the effective interaction with tubulin of some of the prepared compounds and the biological evaluation demonstrate for some of the obtained compounds an interesting activity

4) A protocol to determine microtubule dynamics by quantitative live cell imaging in axon-like processes of model neurons has been established and validated

5) The protocol for the matabolomic studies have been defined

6) two review papers and two original papers have been published. One of them (doi.org/10.1002/chem.202103520 - Fig.1) perfectly described the results obtained for a group of compounds according to the described points 1,2 and 3.

For a detailed description of the performed work, see Technical Report file Part. B
At the moment the network defined the way in which it could arrive from the computational studies till the study of the effect on neurons with some unsolved problems about alpha-synuclein.
The obtained compounds and the subsequent cristallographic studies gave further details about the design of new tubulin binders in maitansin, taxane and pironetin binding site.

We expect to obtain different libraries of:
-tubulin interacting compounds
-molecular probes working at Tau/Tub, alpha-Synuclein/Tub interface
-compounds inhibiting Tau aggregation;
-compounds inhibiting alpha-Synuclein aggregation

We expect to:
-elucidate the contribution of Tau and alpha-Synuclein to modulate MT assembly and dynamics in peripheral neurons and cellular models of neurodegeneration
-validate the effects of selected compounds/peptides on MT system and metabolism in peripheral neurons and cellular models of neurodegeneration, and in Tau and a-Synuclein aggregation in neurons
-validate the effects of novel inhibitors of Tau and α-Synuclein aggregation in neurons

The described experimental work will help us to:
-implement the interdisciplinary knowledge, the soft and transferable skills and the technical skills.
Maytansinol paper - Fig.1
TubInTrain WP1-WP2-WP3 description - Fig.3
TubInTrain general network - Fig.2