Ricerca Progetti nazionali, regionali, internazionaliNAZIONALIPRIN 2022 PNRRTrodusquemine as a novel drug against

Trodusquemine as a novel drug against TDP-43-associated proteinopathies: structural, biological and computational investigation

TAR DNA-binding protein 43 (TDP-43) proteinopathies, including amyotrophic lateral sclerosis (ALS) and frontotemporal dementia (FTD), are neurodegenerative conditions associated with a common histopathology within neurons and glia. This consists in the mislocalization of the TDP-43 from the nucleus to the cytosol, forming well-defined skein-like or round inclusions. The inclusions appeared in both ordered (amyloid-like) or amorphous aggregates. Natural molecules referred to as aminosterols, such as trodusquemine (TRO), have been reported to modulate the aggregation and to suppress the toxicity of α-synuclein protein, related with the onset of Parkinson’s Disease (PD). Recently, aminosterols entered human clinical trials for PD, reporting exciting results.

Given the economical burden that neurological disorders generate in society, there is an urgent need for effective treatments that significantly slow down the neurodegeneration progression. Our data suggests that TRO prevents TDP-43 accumulation and toxicity both in neuronal cells and in a C. elegans model. Thus, the goal of this proposal is to elucidate these protective mechanisms both in vitro and in cultured cells. Thanks to our ability to purify full-length TDP-43, we recently showed a phase separation process, which led rapidly to the formation of apparently round assemblies. We will investigate the influence of TRO in this process by using an array of well-established biophysical methods. Then, the protective effects of TRO will be assessed against the accumulation of both endogenous TDP-43 overexpressed by the cells and exogenous TDP-43, purified in vitro and transfected into the cells. The super-resolution stimulated emission depletion STED microscopy and transmission electron microscopy will be applied to monitor the localization and aggregation state of TDP-43. Raman microspectroscopies in situ will give information about the structure of the TDP-43 aggregates in living cells. Moreover, we will also investigate whether the inhibition of protein tyrosine phosphatase 1B by TRO is involved in its protective effects against TDP-43 neurotoxicity. Finally, we will apply molecular dynamics (MD) simulations to characterize at the atomistic level the aggregation process and the inhibition/recovery of the aggregation performed by TRO. In particular, we will employ Multi-eGO, a structure-based model that has the capabilities to describe the aggregation process at atomic detail. This will allow us to characterize aggregation for TDP-43 alone and in presence of TRO, integrating the experiment performed in the other research units. Taking advantage of our complementary expertises, we will involve three levels of investigation: (1) the biophysical level in vitro, (2) the cell biology in cell cultures, (3) in silico atomistic level analysis. We are confident that this study will provide interesting insights into novel therapeutic opportunities for ALS, FTD and other TDP-43 associated proteinopathies.

 

Data di avvio 30 Novembre 2023

Data di completamento 30 Novembre 2025

Total cost  € 135270

 

Progetto P20225ZPYH finanziato all’interno del Bando PRIN 2022 PNRR di cui al Decreto Direttoriale n. 1409 del 14/9/2022 nell’ambito del Piano Nazionale di Ripresa e Resilienza, Missione 4 – Componente 2. Dalla Ricerca all’Impresa - Investimento 1.1 Fondo per il Programma Nazionale della Ricerca (PNR) e Progetti di Ricerca di Rilevante Interesse Nazionale (PRIN), finanziato dall’Unione europea – NextGenerationEU – CUP B53D23028100001