Development of ?-synuclein-Specific Nanobodies for Targeting Aggregation in Parkinson's Disease

Parkinson's disease (PD) is characterized by the formation of ?-synuclein (?-syn) aggregates, known as Lewy bodies, which contribute to disease progression and neuropathological manifestations. Hence, identifying molecules that can prevent ?-syn aggregation, thereby inhibiting Lewy body formation is of great importance for therapeutic purposes. In our proposed study, we aimed to develop biomolecules, specifically nanobodies (Nbs) that are the antigen?binding domains derived from heavy?chain only antibody. These nanobodies have demonstrated a high affinity for ?-syn fibrils while showing no binding to monomers or other amyloid proteins. The well-characterized nanobodies exhibit several favorable traits, including high affinity, specific activity, and small size. We seek to develop biomolecules that can be used to prevent the formation of ?-syn aggregates. Our research group has developed two novel conformation-specific nanobodies, Nb-04 and Nb-40, which have been selected based on their well-characterized high specificity towards ?-syn fibrils, while not binding to ?-syn monomers or other amyloid proteins. The objective of our study is to assess the effectiveness of these nanobodies in inhibiting the formation of ?-syn aggregates and mitigating the associated toxicity, thus targeting the root cause of PD and related synucleinopathies. To accomplish this, we have defined the following two main aims: Aim 1 – Characterize the in vitro properties of the developed conformation-specific nanobodies, Nb-04 and Nb-40, and evaluate their effectiveness in inhibiting ?-syn aggregates and associated toxicity in cell models. Aim 2 – Assess the efficiency of Nb-04 and Nb-40, in preventing ?-syn aggregation and related neurotoxicity in animal models.

Hamad Bin Khalifa University (HBKU)