Development of Novel Multi-Specific Nanobodies Targeting Alpha-Synuclein Pathology as a Therapeutic Strategy for Parkinson's Disease and other Synucleinopathies

Parkinson’s disease (PD) is a progressive neurodegenerative disorder characterized by the accumulation of misfolded α-synuclein (α-syn) fibrils, which drive neuronal dysfunction and cell death. Current treatments only provide symptomatic relief without targeting disease progression. Our approach involves engineering multi-specific nanobodies (Nbs) to selectively target and neutralize diverse α-syn fibril strains, potentially offering a disease-modifying therapy for PD. Our preliminary findings demonstrate that our second generation monovalent Nbs (Nb-04 and Nb-40) exhibit high specificity for α-syn fibrils with beta-sheet structure. To enhance the disease coverage and to capture the maximum spectrum of α-syn fibrils, we designed bispecific and trispecific Nbs with improved binding affinity, avidity, and specificity, while preserving the unique characteristics of the Nbs. We will characterize these Nbs biochemically and biophysically using filter retardation assay, surface plasmon resonance (SPR), ELISA-based epitope mapping, and α-syn aggregation inhibition assays. Functional validation will be conducted in PD cell models and in vivo PFFs PD mouse models to assess neuroprotection. This study aims to generate multi-specific Nbs capable of inhibiting the pathological spread of different strains of α-syn aggregates simultaneously, offering a promising strategy for halting or slowing PD progression.

Hamad Bin Khalifa University (HBKU)