The chaperone HSPB8 reduces the accumulation of truncated TDP-43 species in cells and protects against TDP-43-mediated toxicity

• Published in: Human molecular genetics Vol. 25; no. 18; pp. 3908 - 3924
• Main Authors: Crippa, Valeria, Cicardi, Maria Elena, Ramesh, Nandini, Seguin, Samuel J, Ganassi, Massimo, Bigi, Ilaria, Diacci, Chiara, Zelotti, Elena, Baratashvili, Madina, Gregory, Jenna M, Dobson, Christopher M, Cereda, Cristina, Pandey, Udai Bhan, Poletti, Angelo, Carra, Serena
• Format: Journal Article
• Language: English
• Published: England Oxford University Press 15-09-2016
• Subjects: Amyotrophic Lateral Sclerosis - genetics; Amyotrophic Lateral Sclerosis - pathology; Animals; Disease Models, Animal; DNA-Binding Proteins - genetics; DNA-Binding Proteins - metabolism; Drosophila melanogaster - genetics; Drosophila Proteins - biosynthesis; Drosophila Proteins - genetics; Eye - growth & development; Eye - physiopathology; Gene Expression Regulation; Heat-Shock Proteins - biosynthesis; Heat-Shock Proteins - genetics; Humans; Mice; Mice, Transgenic; Molecular Chaperones; Motor Neurons - metabolism; Motor Neurons - pathology; Peptide Fragments - genetics; Peptide Fragments - metabolism; Protein Aggregation, Pathological - genetics; Protein Serine-Threonine Kinases - biosynthesis; Protein Serine-Threonine Kinases - genetics; Pupa - genetics; Pupa - growth & development
• ISSN: 0964-6906; 1460-2083
• DOI: 10.1093/hmg/ddw232

Aggregation of TAR-DNA-binding protein 43 (TDP-43) and of its fragments TDP-25 and TDP-35 occurs in amyotrophic lateral sclerosis (ALS). TDP-25 and TDP-35 act as seeds for TDP-43 aggregation, altering its function and exerting toxicity. Thus, inhibition of TDP-25 and TDP-35 aggregation and promotion of their degradation may protect against cellular damage. Upregulation of HSPB8 is one possible approach for this purpose, since this chaperone promotes the clearance of an ALS associated fragments of TDP-43 and is upregulated in the surviving motor neurones of transgenic ALS mice and human patients. We report that overexpression of HSPB8 in immortalized motor neurones decreased the accumulation of TDP-25 and TDP-35 and that protection against mislocalized/truncated TDP-43 was observed for HSPB8 in Drosophila melanogaster Overexpression of HSP67Bc, the functional ortholog of human HSPB8, suppressed the eye degeneration caused by the cytoplasmic accumulation of a TDP-43 variant with a mutation in the nuclear localization signal (TDP-43-NLS). TDP-43-NLS accumulation in retinal cells was counteracted by HSP67Bc overexpression. According with this finding, downregulation of HSP67Bc increased eye degeneration, an effect that is consistent with the accumulation of high molecular weight TDP-43 species and ubiquitinated proteins. Moreover, we report a novel Drosophila model expressing TDP-35, and show that while TDP-43 and TDP-25 expression in the fly eyes causes a mild degeneration, TDP-35 expression leads to severe neurodegeneration as revealed by pupae lethality; the latter effect could be rescued by HSP67Bc overexpression. Collectively, our data demonstrate that HSPB8 upregulation mitigates TDP-43 fragment mediated toxicity, in mammalian neuronal cells and flies.