Abstract
SummaryThe Parkinson's disease-associated gene, LRRK2, is also associated with immune disorders and infectious disease and is expressed in immune subsets. Here, we characterize a platform for interrogating the expression and function of endogenous LRRK2 in authentic human phagocytes using human induced pluripotent stem cell-derived macrophages and microglia. Endogenous LRRK2 is expressed and upregulated by interferon-γ in these cells, including a 187-kDa cleavage product. Using LRRK2 knockout and G2019S isogenic repair lines, we find that LRRK2 is not involved in initial phagocytic uptake of bioparticles but is recruited to LAMP1+/RAB9+ “maturing” phagosomes, and LRRK2 kinase inhibition enhances its residency at the phagosome. Importantly, LRRK2 is required for RAB8a and RAB10 recruitment to phagosomes, implying that LRRK2 operates at the intersection between phagosome maturation and recycling pathways in these professional phagocytes.
Highlights
LRRK2 encodes a large (286 kDa), multi-domain cytoplasmic protein, with both guanosine triphosphatase (GTPase) and kinase domains, flanked by several protein-protein interaction domains
We show that LRRK2 is expressed in human induced pluripotent stem cells (hiPSCs) macrophages and microglia, with expression significantly upregulated by interferon-g (IFN-g) and identify the cleavage region of a truncated LRRK2 product found in this lineage
IFN-g treatment significantly upregulated the percentage of LRRK2 expressing hiPSC microglia to 86%, its expression level plateauing by 48 h post IFN-g treatment (Figure 2D). These results demonstrate the validity of the hiPSC macrophage and microglia models for investigating endogenous LRRK2 function
Summary
LRRK2 (leucine-rich repeat kinase 2) encodes a large (286 kDa), multi-domain cytoplasmic protein, with both guanosine triphosphatase (GTPase) and kinase domains, flanked by several protein-protein interaction domains. Mutations in LRRK2 account for approximately 4% of familial and 1% of idiopathic cases of the progressive neurodegenerative disorder, Parkinson’s disease (PD), forming an important genetic risk factor for PD. LRRK2 variants are associated with autoimmune disorders (Witoelar et al, 2017), Crohn’s disease, and with infectious diseases, notably Mycobacterium leprae (Wang et al, 2015; Zhang et al, 2009). LRRK2 expression has been linked with Mycobacterium tuberculosis infection (Hartlova et al, 2018; Wang et al, 2018). LRRK2 is expressed in a variety of cell lineages, including several immune subsets, notably B cells, neutrophils, monocytes, macrophages, and microglia (Atashrazm et al, 2019; Fan et al, 2018; Gardet et al, 2010; Hakimi et al, 2011; Kim et al, 2012; Marker et al, 2012; Moehle et al, 2012; Thevenet et al, 2011; reviewed in Lee et al, 2017)
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