α-Synuclein fibril-specific nanobody reduces prion-like α-synuclein spreading in mice

• Published in: Nature communications 2022-07, Vol.13 (1), p.4060-13, Article 4060
• Main Authors: Butler, Yemima R., Liu, Yuqing, Kumbhar, Ramhari, Zhao, Peiran, Gadhave, Kundlik, Wang, Ning, Li, Yanmei, Mao, Xiaobo, Wang, Wenjing
• Format: Article
• Language: English
• Subjects: 101/28; 13/1; 13/109; 13/31; 14/19; 38/35; 38/70; 38/77; 42/40; 631/378/1689/364; 631/92/469; 64/60; 82/16; 82/80; 82/83; alpha-Synuclein - genetics; alpha-Synuclein - metabolism; Animal models; Animals; Dementia disorders; Fibrils; Humanities and Social Sciences; Humans; Immunoreactivity; Lewy bodies; Lewy Body Disease; Mice; Monomers; Movement disorders; multidisciplinary; Nanobodies; Neurodegenerative diseases; Parkinson Disease - metabolism; Parkinson's disease; Pathogenesis; Pathology; Phosphorylation; Prion protein; Prions; Science; Science (multidisciplinary); Synuclein; Synucleinopathies; Transgenic mice; Viruses; Yeasts
• ISSN: 2041-1723; 2041-1723
• DOI: 10.1038/s41467-022-31787-2

Pathogenic α-synuclein (α-syn) is a prion-like protein that drives the pathogenesis of Lewy Body Dementia (LBD) and Parkinson’s Disease (PD). To target pathogenic α-syn preformed fibrils (PFF), here we designed extracellular disulfide bond-free synthetic nanobody libraries in yeast. Following selection, we identified a nanobody, PFFNB2, that can specifically recognize α-syn PFF over α-syn monomers. PFFNB2 cannot inhibit the aggregation of α-syn monomer, but can significantly dissociate α-syn fibrils. Furthermore, adeno-associated virus (AAV)-encoding EGFP fused to PFFNB2 (AAV-EGFP-PFFNB2) can inhibit PFF-induced α-syn serine 129 phosphorylation (pS129) in mouse primary cortical neurons, and prevent α-syn pathology spreading to the cortex in the transgenic mice expressing human wild type (WT) α-syn by intrastriatal-PFF injection. The pS129 immunoreactivity is negatively correlated with the expression of AAV-EGFP-PFFNB2. In conclusion, PFFNB2 holds a promise for mechanistic exploration and therapeutic development in α-syn-related pathogenesis. Butler et al. selected disulfide bond-free nanobodies to target α-synuclein fibrils. Nanobody PFFNB2 was shown to disaggregate α-synuclein fibrils in vitro and inhibit α-synuclein pathology development in neuron cultures and mouse models.