Abstract
Variants in the LRRK2 and GBA1 genes are among the most common risk factors associated with Parkinson’s disease (PD). Both patients carrying PD-associated variants in GBA1, encoding lysosomal enzyme glucocerebrosidase (GCase), and a subset of non-carrier patients have been shown to have reduced GCase enzymatic activity, suggesting that reduced GCase activity may be a feature of both genetic and a subset of sporadic PD. However, the effect of PD-associated variants in LRRK2, encoding a serine/threonine kinase, on GCase activity remains controversial, with conflicting results in various tissues and cell types. Moreover, rare patients carrying both GBA1 and LRRK2 risk alleles seem to have a more benign disease course than carriers of GBA1 variants alone, suggesting a complex interplay between these two genes in PD. Here, we evaluate the effect of LRRK2 kinase activity on GCase activity in human induced pluripotent stem cell (iPSC)-derived microglia (iMGs), a PD-relevant brain cell type expressing high levels of LRRK2. Using CRISPR editing, isogenic control iPSC lines were generated to match PD patient-derived iPSC lines harbouring the LRRK2 p.G2019S, p.M1646T, or p.N551K-p.R1398H protective haplotype variants. Whereas iMGs harbouring the p.M1646T variant, and the protective haplotype, respectively increased and decreased phosphorylation of canonical LRRK2 substrate, Rab10, GCase protein levels and activity were not altered in any of the LRRK2 variant lines. Additionally, whereas pharmacological inhibition of LRRK2 kinase activity had no impact on GCase activity in iMGs under basal conditions, it attenuated the increase in GCase activity elicited in response to interferon γ (IFNγ) treatment. Moreover, GCase activity induced by IFNγ was reduced in PD risk LRRK2 p.M1646T iMGs and increased in p.N551K-p.R1398H protective haplotype iMGs compared to their isogenic corrected controls, congruent with their respective effects on LRRK2 kinase activity and PD risk. Thus, our data suggest a role for LRRK2 kinase activity in regulation of GCase activity in response to neuroinflammation.
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Acknowledgements
Thanks to Wolfgang Reintsch and Julien Sirois for training and support with high content imaging and flow cytometry experiments, respectively. Thanks to Marie-France Dorion for training and advice on differentiation and characterization of iPSC-derived microglia. Thanks to Jace Jones-Tabah, Roxanne Larivière, and Andrea Krahn for establishing and optimizing live-cell GCase, lysosomal proteolytic, and pH assays. E.J.M. has been supported by a Fonds de Recherche du Québec-Santé Doctoral Fellowship and Jeanne Timmins Costello Fellowship awarded by the Integrated program in Neuroscience at McGill University. This work was funded by grants from The Michael J. Fox Foundation for Parkinson’s Research (MJFF-019045) and from the Canadian Institutes of Health Research (FDN-154301). G-Can (GBA1 Canada) Initiative, an open-science collaborative initiative aimed at addressing GBA1 associated neurodegeneration, has contributed to this research. G-Can is supported by The Hilary & Galen Weston Foundation, J. Sebastian van Berkom, Ghislaine Saucier and Silverstein Foundation. E.A.F. is supported by a Canada Research Chair (Tier 1) in Parkinson’s disease.
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E.J.M. generated p.M1646T and protective haplotype isogenic correction cell lines, differentiated iMGs, designed and performed experiments, analysed data, and prepared the manuscript. C.X-.Q.C. and N.A. performed quality control experiments on iPSC lines used in this study. E.D. designed CRISPR editing strategies and ddPCR screening strategies to generate isogenic correction lines, and generated the p.G2019S isogenic correction line. Z.Y. designed CRISPR KO strategy and generated the LKO line. D.K. performed xMAP experiments and analysed data. T.M.D., K.S., and Z.G-.O. designed experiments. E.A.F. designed experiments, supervised the project, and prepared the manuscript.
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MacDougall, E.J., Chen, C.XQ., Deneault, E. et al. LRRK2 kinase mediates increased GCase activity in microglia in response to IFNγ-induced proinflammatory stimulation. npj Parkinsons Dis. (2026). https://doi.org/10.1038/s41531-026-01310-1
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DOI: https://doi.org/10.1038/s41531-026-01310-1
