Arisla | The research

Active project: RBPALS

RBPALS:

Characterization of disease mechanisms mediated by TDP-43 and FUS RNA-binding proteins in Amyotrophic Lateral Sclerosis

The Project

The identification of TDP-43 protein as a main component of the intracellular inclusions observed in patients affected from both sporadic and SOD1-negative familial forms of ALS has opened up new perspectives with regards to the potential pathogenic clues for this disease. The genetic findings of causative mutations in TARDBP and, recently, in FUS genes further suggest that post-transcriptional regulatory processes have an important role in ALS pathogenesis. The TDP-43 and FUS proteins, in fact, are nuclear RNA-binding proteins (RBP) mainly associated with splicing activities, but also implicated in mRNA stabilization and transport. Since in ALS-affected tissues both these RBPs are mislocalized in the cytoplasm where they form aggregates, the “loss-“ versus the “gain-of-function” hypotheses represent an important issue to be addressed for future drug-development prospects and preclinical approaches. Our project aims to provide experimental data regarding the biological role played by TDP-43 and FUS RBPs in neuronal cells both in normal and disease states. In order to establish which scenario better accounts for the neurodegenerative processes observed in ALS patients, genes and eventually pathways, whose expression is affected by TDP-43 and FUS dysfunctions, will be characterized. The effects of TDP-43/FUS depletion from the cellular environment will be assessed using microarray technology for gene expression and splicing profilings. We will include an accurate characterization of TDP-43 and FUS intermolecular interactions and biological properties using a variety of techniques (RNA immunoprecipitation and chip, CLIP and mass-spec analyses). Finally, we propose the establishment and characterization of “gain/loss-of-function” cell models through the selective expression of mutant TDP-43 proteins and concomitant gene silencing of the endogenous wild-type form. These cell lines, and the Drosophila “loss-of-function” model for TDP-43 available in Prof. Baralle’s lab, will represent valuable tools for drug discovery and to test therapeutical strategies common to sporadic and SOD1-negative familial forms.

The Research Team

Principal Investigator:

RATTI