Nanobodies against LRRK2: a new tool for Parkinson's research

Parkinson’s disease is a common neurodegenerative disorder characterized by the progressive loss of dopaminergic neurons, which causes motor and non-motor symptoms. Several studies highlighted the key role of leucine-rich repeat kinase 2 (LRRK2), an enzyme involved in various cellular functions. Importantly, mutations in its gene result in abnormal kinase activity, leading to Parkinson’s pathogenesis. Therefore, a strategy to modulate its activity is strongly pursued by research, and various inhibitors and interactors have already been discovered. In particular, nanobodies (Nbs) are versatile compounds which bind to LRRK2 generating multiple effects. This project aims to explore the potential of Nbs as a new tool to study the kinase in BV2 microglia cells in distinct treatment conditions. After Nbs transfection, two compounds, MLi-2 and chloroquine, have been used to trigger the relocation of cytoplasmic LRRK2 into different subcellular structures. Through immunocytochemistry and confocal microscopy, co-localization between Nbs and the kinase has been analyzed. As a result, the experiments showed that Nbs could represent a valid strategy to track LRRK2, although some unexpected outcomes occurred, probably due to aspecific interactions or background noise in microscope acquisitions. However, the project highlights the potential of Nbs even for the development of targeted therapies for Parkinson’s disease.