Data di Pubblicazione:
2023
Citazione:
In vivo functional validation of the ALS causative p.E696K missense mutation in TBK1 / I. Raya, Y. Douahem, D. Brenner, R. Yilmaz, V. Ast, A. Duda, Q. Gao, R.M. Cristofani, A. Poletti, V. Maglione, J. Weishaupt, R. Parlato. ((Intervento presentato al convegno ENCALS meeting tenutosi a Barcelona : 12-14 luglio nel 2023.
Abstract:
Deleterious mutations in the autophagy regulator TANK1-binding kinase 1 (TBK1) cause ALS. Unlike other ALS causa- tive TBK1 missense mutations, p.E696K selectively abolishes the interaction of the TBK1 protein with the autophagy adaptor protein optineurin (OPTN). The p.E696K mutation triggers ALS most likely through a partial loss-of-function. Our group has generated Tbk1E696K knock-in mice carrying the p.E696K mutation leading to a block in the autophagic flux, while kinase activity is preserved. The mutant mice show age-dependent ALS-like motor and neuropathological phenotypes, such as the increased accumulation of cytosolic p62 positive inclusions. In this study, we apply these models for: 1) a better understanding of the cellular and molecular dysfunction linked to the onset and progression of mutant TBK1 (mTBK1) pathology, and 2) functionally validating the impact of mTBK1 on the accumulation of toxic proteins. To this end, we have investigated how mTBK1 dysregulates transcriptional profiles of different cell-types in the spinal cord of the Tbk1E696K knock-in mice at pre- and early-symptomatic stages by single nuclei RNA-sequen- cing. In parallel, we have generated double knock-in mice carrying a mutant Huntingtin (mHTT) allele in combination with the Tbk1E696K knock-in. Based on previous evidence that HTT is phosphorylated by TBK1, we have analyzed the impact of Tbk1E696K on accumulation of mHTT protein inclusions in different brain regions. In particular, we have found early transcriptional changes in astrocyte and microglia profiles. Moreover, we found a consistent increase in the number of mHTT and p62 positive inclusions in the double mutant mice, supporting that the p.E696K mutation can impair clearance of aggregation prone proteins. These studies suggest the potential application of these mo- dels to dissect the cell-, stage-, and disease-specific function of TBK1, and to possibly identify shared across disease mechanisms.
Tipologia IRIS:
14 - Intervento a convegno non pubblicato
Elenco autori:
I. Raya, Y. Douahem, D. Brenner, R. Yilmaz, V. Ast, A. Duda, Q. Gao, R.M. Cristofani, A. Poletti, V. Maglione, J. Weishaupt, R. Parlato
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