LRRK2 mediates tubulation and vesicle sorting from lysosomes

Bonet-Ponce, Luis; Beilina, Alexandra; Williamson, Chad D.; Lindberg, Eric; Kluss, Jillian H.; Sáez-Atiénzar, Sara; Landeck, Natalie; Kumaran, R. Ileng; Mamais, Adamantios; Bleck, Christopher K. E.; Li, Yan; Cookson, Mark

doi:10.1126/sciadv.abb2454

Cited by 170 publications

(235 citation statements)

References 63 publications

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“…Second, we were able to control the activation state of cells, which again would be impossible to perform in the human brain. Here, we extend our recent observation that lysosomal damage can activate the LRRK2 signaling pathway (24) to show that activation can occur in iMicroglia and that activation shows interaction with genotype. Given results from other laboratories that inflammatory signaling can promote LRRK2 activation through a similar pathway (30), we infer that neuroinflammation may interact with genotype at LRRK2, and potentially at other lysosomal genes associated with PD (31), to influence overall brain phenotype.…”

Section: The Rs76904798 Variant Does Not Drive the Observed Lrrk2 Eqtsupporting

confidence: 80%

“…In this cell model, we could measure LRRK2 kinase activity based on the phosphorylation of physiological substrate Rab10 (23). As well as baseline activity, we explored methods of microglia stimulation and LRRK2 activation by exposing cells for two hours to lipopolysaccharide (LPS), α-synuclein conditioned medium (aSCM), or L-leucyl-L-leucine methyl ester (LLOMe), which we have recently shown to activate LRRK2 in primary mouse astrocytes (24). Immunoblot analysis indicated that of the three treatments, LLOMe most strongly activated LRRK2 kinase activity in iMicroglia as measured by significantly increased phosphorylation of Rab10 ( Fig.…”

Section: Ipsc-derived Microglia Are Transcriptionally Similar To Braimentioning

confidence: 99%

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Association of a Common Genetic Variant with Parkinson’s Disease is Propagated through Microglia

Langston

Beilina

Reed

et al. 2021

Preprint

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Studies of the genetic basis of Parkinson’s disease (PD) have identified many disease-associated genetic variants, but the mechanisms linking variants to pathogenicity are largely unknown. PD risk is attributed to both coding mutations in the Leucine-rich repeat kinase 2 (LRRK2) gene and to common non-coding variation upstream of the LRRK2 locus. Here we show that the influence of genotype at non-coding variant rs76904798 on LRRK2 expression is propagated specifically through microglia, in contrast to evaluations based on general rather than genotype-dependent expression. We find evidence of microglia-specific regulatory regions that may modulate LRRK2 expression using single nuclei sequencing analyses of human frontal cortex and confirm these results in a human induced pluripotent stem cell-derived microglia model. Our study demonstrates that cell type is an important consideration in interrogation of the role of non-coding variation in disease pathogenesis.

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Section: The Rs76904798 Variant Does Not Drive the Observed Lrrk2 Eqtsupporting

confidence: 80%

Section: Ipsc-derived Microglia Are Transcriptionally Similar To Braimentioning

confidence: 99%

Association of a Common Genetic Variant with Parkinson’s Disease is Propagated through Microglia

Langston

Beilina

Reed

et al. 2021

Preprint

Self Cite

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“…It would be interesting to explore whether these agonists promote the recruitment of LRRK2 to particular membrane compartments, thereby activating LRRK2 and promoting Rab10 and Rab12 phosphorylation at that location. Recruitment of LRRK2 to membranes has been proposed previously to be a key mechanism by which LRRK2 activity is regulated [ 29 , 57 , 68 ].…”

Section: Discussionmentioning

confidence: 99%

Endogenous Rab29 does not impact basal or stimulated LRRK2 pathway activity

Kalogeropulou

Freemantle

Lis

et al. 2020

Biochemical Journal

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Mutations that enhance LRRK2 protein kinase activity cause inherited Parkinson’s disease. LRRK2 phosphorylates a group of Rab GTPase proteins, including Rab10 and Rab12, within the effector-binding switch-II motif. Previous work has indicated that the PARK16 locus, which harbors the gene encoding for Rab29, is involved in Parkinson's, and that Rab29 operates in a common pathway with LRRK2. Co-expression of Rab29 and LRRK2 stimulates LRRK2 activity by recruiting LRRK2 to the surface of the trans Golgi network. Here we report that knock-out of Rab29 does not influence endogenous LRRK2 activity, based on assessment of Rab10 and Rab12 phosphorylation, in wildtype LRRK2, LRRK2[R1441C] or VPS35[D620N] knock-in mouse tissues and primary cell lines, including brain extracts and embryonic fibroblasts. We find that in brain extracts, Rab12 phosphorylation is more robustly impacted by LRRK2 inhibitors and pathogenic mutations than Rab10 phosphorylation. Transgenic overexpression of Rab29 in a mouse model was also insufficient to stimulate basal LRRK2 activity. We observed that stimulation of Rab10 and Rab12 phosphorylation induced by agents that stress the endolysosomal system (nigericin, monensin, chloroquine, and LLOMe) was not blocked in Rab29 deficient mouse embryonic fibroblasts. From the agents tested, nigericin induced the greatest increase in pRab10 and pRab12 phosphorylation (5-9 fold). Our findings indicate that basal, pathogenic, as well as nigericin and monensin stimulated LRRK2 pathway activity is not controlled by Rab29. Further work is required to establish how LRRK2 activity is regulated, and whether other Rab proteins can control LRRK2 by targeting it to diverse membranes.

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“…Previous studies have shown that pharmacological manipulations that induce lysosomal damage trigger LRRK2 translocation to lysosomes and lead to an increase in Rab phosphorylation (26,27). Further, LRRK2 expression in immune cells is elevated in response to various inflammatory stimuli such as IFN- (28)(29)(30).…”

Section: High Lrrk2 Expression and Activity Are Observed In Mouse Glimentioning

confidence: 99%

Understanding LRRK2 kinase activity in preclinical models and human subjects through quantitative analysis of LRRK2 and pRab10

Wang

Negrou

Maloney

et al. 2021

Preprint

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Variants in the leucine-rich repeat kinase 2 (LRRK2) gene are associated with increased risk for familial and sporadic Parkinson's disease (PD). Pathogenic variants in LRRK2, including the common variant G2019S, result in increased LRRK2 kinase activity, supporting the therapeutic potential of LRRK2 kinase inhibitors for PD. To better understand the role of LRRK2 in disease and to support the clinical development of LRRK2 inhibitors, quantitative and high-throughput assays to measure LRRK2 levels and activity are needed. We developed and applied such assays to measure the levels of LRRK2 as well as the phosphorylation of LRRK2 itself or one of its substrates, Rab10 (pT73 Rab10). We observed increased LRRK2 activity in various cellular models of disease, including iPSC-derived microglia, as well as in human subjects carrying disease-linked variant in LRRK2 (G2019S). Capitalizing on the high-throughput and sensitive nature of these assays, we detected a significant reduction in LRRK2 activity in subjects carrying missense variants in LRRK2 associated with reduced disease risk. Finally, we optimized these assays to enable analysis of LRRK2 activity following inhibition in human peripheral blood mononuclear cells (PBMCs) and whole blood, demonstrating their potential utility as biomarkers to assess changes in LRRK2 expression and activity in the clinic.

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LRRK2 mediates tubulation and vesicle sorting from lysosomes

Cited by 170 publications

References 63 publications

Association of a Common Genetic Variant with Parkinson’s Disease is Propagated through Microglia

Association of a Common Genetic Variant with Parkinson’s Disease is Propagated through Microglia

Endogenous Rab29 does not impact basal or stimulated LRRK2 pathway activity

Understanding LRRK2 kinase activity in preclinical models and human subjects through quantitative analysis of LRRK2 and pRab10

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