Objective: The aim was to evaluate the effectiveness of leucine-rich repeat kinase 2 (LRRK2) kinase activity inhibition in restoring transport of the enzyme glucocerebrosidase (GCase), encoded by the GBA gene, to the lysosomes in peripheral blood primary monocytes-derived macrophages (PMDMs) of patients with GBA-associated Parkinson’s disease (GBA-PD).
Background: Mutations in the GBA gene lead to reduced GCase activity and are the most common genetic risk factors for Parkinson’s disease (PD). Recent data revealed, that inhibition of kinase activity of LRRK2 may influence GCase activity [1, 2, 3].
Method: Peripheral blood monocytes (MDMs) of 5 GBA‐PD patients and 6 controls were cultured with MCS-F (10ng/ml) for 5 days. After differentiation MDMs were cultured for 4 days in the presence (treated) or absence (untreated) of selective inhibitor of kinase activity LRRK2, MLi-2 (100nM/ml). Colocalization of GCase with the lysosomal marker LAMP-2 was assessed by confocal microscopy (Leica TCS‐SP5). GCase activity and level was measured by liquid chromatography–mass spectrometry and western blot analysis, respectively.
Results: Inhibition of LRRK2 kinase activity increased GCase colocalization with LAMP2 in GBA‐PD and controls PMDMscompared to untreated cells (p<0.0001). As expected, GCase activity was decreased in untreated of GBA-PD compared to untreated PMDMs of controls (p<0.05). Inhibition of LRRK2 kinase activity increased protein level and activity of GCase in treated PMDMs compared to untreated in GBA-PD patients (p<0.05). Interestingly, GCase activity in treated PMDMs of GBA-PD patients reached activity level observed in untreated PMDMs of controls. A tendency towards increased GCase activity was observed in treated compared to untreated PMDMs of controls (p>0.05).
Conclusion: We demonstrated, that inhibition of LRRK2 kinase activity restores GCase transport to lysosomes and enhances GCase level and activity in GBA-PD PMDMs. Our results provide insights into the interaction of LRRK2 and GCase.
This study was supported by RSF №22-25-00501.
References: [1] Ysselstein D, et al. LRRK2 kinase activity regulates lysosomal glucocerebrosidase in neurons derived from Parkinson’s disease patients. Nat Commun. 2019 Dec 5;10(1):5570.
[2] Sanyal A, et al. LRRK2 Kinase Inhibition Rescues Deficits in Lysosome Function Due to Heterozygous GBA1 Expression in Human iPSC-Derived Neurons. Frontiers in Neuroscience. 2020; 14:442.
[3] Kedariti M., et al. LRRK2 kinase activity regulates GCase level and enzymatic activity differently depending on cell type in Parkinson’s disease. npj Parkinsons Dis. 2022; 8, 92.
To cite this abstract in AMA style:
K. Basharova, A. Bezrukova, A. Izyumchenko, G. Baydakova, I. Miliukhina, E. Zakharova, S. Pchelina, T. Usenko. Inhibition of LRRK2 kinase activity promotes glucocerebrosidase transport to the lysosomes in primary monocyte-derived macrophages from patients with GBA-associated Parkinson’s disease [abstract]. Mov Disord. 2023; 38 (suppl 1). https://www.mdsabstracts.org/abstract/inhibition-of-lrrk2-kinase-activity-promotes-glucocerebrosidase-transport-to-the-lysosomes-in-primary-monocyte-derived-macrophages-from-patients-with-gba-associated-parkinsons-disease/. Accessed November 21, 2024.« Back to 2023 International Congress
MDS Abstracts - https://www.mdsabstracts.org/abstract/inhibition-of-lrrk2-kinase-activity-promotes-glucocerebrosidase-transport-to-the-lysosomes-in-primary-monocyte-derived-macrophages-from-patients-with-gba-associated-parkinsons-disease/