Both
leucine-rich repeat
kinase 2 (LRRK2) and
glucocerebrosidase (GCase) are promising targets for the treatment of
Parkinson's disease (PD). Evidence suggests that both
proteins are involved in biological pathways involving the lysosome. However, studies to date have largely investigated the
enzymes in isolation and any relationship between LRRK2 and GCase remains unclear. Both
enzymes are highly expressed in peripheral blood monocytes and have been implicated in immune function and
inflammation. To facilitate the standardized measurement of these readouts in large cohorts of samples collected from persons with PD across the globe, we developed and optimized a sample collection and processing protocol with parallel flow cytometry assays. Assay parameters were first optimized using healthy control peripheral blood mononuclear cells (PBMCs), and then LRRK2 and GCase activities were measured in immune cells from persons with idiopathic PD (iPD). We tested the ability of this protocol to deliver similar results across institutes across the globe, and named this protocol the Wallings-Hughes Optimized Protocol for PBMC Assessment (WHOPPA). In the application of this protocol, we found increased LRRK2 levels and stimulation-dependent enzymatic activity, and decreased GBA index in classical iPD monocytes, as well as increased
cytokine release in PD PBMCs. WHOPPA also demonstrated a strong positive correlation between LRRK2 levels, pRab10 and
HLA-DR in classical monocytes from subjects with iPD. These data support a role for the global use of WHOPPA and expression levels of these two PD-associated
proteins in immune responses, and provide a robust assay to determine if LRRK2 and GCase activities in monocytes have potential utility as reliable and reproducible
biomarkers of disease in larger cohorts of subjects with PD.