LRRK2 mediates tubulation and vesicle sorting from membrane damaged lysosomes

Bonet-Ponce, Luis; Beilina, Alexandra; Cd, Williamson; Lindberg, Eric; Jh, Kluss; Sáez-Atiénzar, Sara; Landeck, Natalie; Kumaran, R. Ileng; Mamais, Adamantios; Bleck, Christopher K. E.; Y, Li; Mr, Cookson

doi:10.1101/2020.01.23.917252

Cited by 15 publications

(14 citation statements)

References 56 publications

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“…To further test this hypothesis, we treated cells with the lysosomal destabilizing agent LLOMe that interacts with the lysosomal membrane and luminal hydrolases. Using super-resolution microscopy, we found that LLOMe induced recruitment of WT LRRK2 to the membrane of enlarged lysosomes as previously shown (17) (Fig 2C). Endogenous Rab8a was also recruited to the lysosomal membrane in WT LRRK2 cells after treatment with LLOMe ( Fig 2D).…”

Section: Pathogenic Mutants Of Lrrk2 Sequester Rab8a To Damaged Lysossupporting

confidence: 86%

“…Previous studies have shown that LRRK2 can localize to enlarged lysosomes along with Rab GTPases (18). Furthermore, we have shown that LRRK2 can be recruited to damaged lysosomes that have low degradative capacity (17). In the current model system where we see mutant LRRK2 recruiting Rab8a to Lamp1/Lamp2-positive lysosomes, those LRRK2 structures were found to be Cathepsin D negative (Fig 2A and 2B for Airyscan images).…”

Section: Pathogenic Mutants Of Lrrk2 Sequester Rab8a To Damaged Lysossupporting

confidence: 58%

“…LRRK2 can phosphorylate Rab8 and Rab10, and reports suggest downstream effects on lysosomal integrity. We, and others, have reported phosphorylationdependent recruitment of Rab10 onto damaged lysosomes by LRRK2, in a process that orchestrates lysosomal homeostasis (17,18).…”

Section: Introductionmentioning

confidence: 73%

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Pathogenic mutations in LRRK2 sequester Rab8a to damaged lysosomes and regulate transferrin-mediated iron uptake in microglia

Mamais

Landeck

Langston

et al. 2020

Preprint

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Mutations in leucine-rich repeat kinase 2 (LRRK2) cause autosomal dominant Parkinson’s disease (PD) while polymorphic LRRK2 variants are associated with sporadic PD. PD-linked mutations increase LRRK2 kinase activity and induce neurotoxicity in vitro and in vivo. The small GTPase Rab8a is a LRRK2 kinase substrate and is involved in receptor-mediated recycling and endocytic trafficking of transferrin, but the effect of PD-linked LRRK2 mutations on the function of Rab8a are poorly understood. Here, we show that gain-of-function mutations in LRRK2 induce sequestration of endogenous Rab8a into lysosomes in cells while pharmacological inhibition of LRRK2 kinase activity reverses this phenotype. Furthermore, we show that LRRK2 mutations drive accumulation of endocytosed transferrin into Rab8a-positive lysosomes leading to a dysregulation of iron transport. LRRK2 has been nominated as an integral part of cellular responses downstream of proinflammatory signals and is activated in microglia in post-mortem PD tissue. Here, we show that iPSC-derived microglia from patients carrying the most common LRRK2 mutation, G2019S, mistraffic transferrin to lysosomes proximal to the nucleus in proinflammatory conditions. Furthermore, G2019S knock-in mice show significant increase in iron deposition in microglia following intrastriatal LPS injection compared to wild type mice, accompanied by striatal accumulation of ferritin. Our data support a role of LRRK2 in modulating iron uptake and storage in response to proinflammatory stimuli in microglia.

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Section: Pathogenic Mutants Of Lrrk2 Sequester Rab8a To Damaged Lysossupporting

confidence: 86%

Section: Pathogenic Mutants Of Lrrk2 Sequester Rab8a To Damaged Lysossupporting

confidence: 58%

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Pathogenic mutations in LRRK2 sequester Rab8a to damaged lysosomes and regulate transferrin-mediated iron uptake in microglia

Mamais

Landeck

Langston

et al. 2020

Preprint

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“…The consequence of a chronic RAB10 hyperphosphorylation could result in reduced ciliogenesis, thus impacting on brain functionality as suggested by recent works [ 43 , 44 ]. Future studies addressing the precise consequences of R1441C-dependent RAB10 hyperphosphorylation on ciliogenesis, as well as on lysosomal function [ 41 , 45 ], will better clarify the pathogenic mechanisms of this mutation.…”

Section: Discussionmentioning

confidence: 99%

Divergent Effects of G2019S and R1441C LRRK2 Mutations on LRRK2 and Rab10 Phosphorylations in Mouse Tissues

Iannotta

Biosa

Kluss

et al. 2020

Cells

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Mutations in LRRK2 cause familial Parkinson’s disease and common variants increase disease risk. LRRK2 kinase activity and cellular localization are tightly regulated by phosphorylation of key residues, primarily Ser1292 and Ser935, which impacts downstream phosphorylation of its substrates, among which Rab10. A comprehensive characterization of LRRK2 activity and phosphorylation in brain as a function of age and mutations is missing. Here, we monitored Ser935 and Ser1292 phosphorylation in midbrain, striatum, and cortex of 1, 6, and 12 months-old mice carrying G2019S and R1441C mutations or murine bacterial artificial chromosome (BAC)-Lrrk2-G2019S. We observed that G2019S and, at a greater extent, R1441C brains display decreased phospho-Ser935, while Ser1292 autophosphorylation increased in G2019S but not in R1441C brain, lung, and kidney compared to wild-type. Further, Rab10 phosphorylation, is elevated in R1441C carrying mice, indicating that the effect of LRRK2 mutations on substrate phosphorylation is not generalizable. In BAC-Lrrk2-G2019S striatum and midbrain, Rab10 phosphorylation, but not Ser1292 autophosphorylation, decreases at 12-months, pointing to autophosphorylation and substrate phosphorylation as uncoupled events. Taken together, our study provides novel evidence that LRRK2 phosphorylation in mouse brain is differentially impacted by mutations, brain area, and age, with important implications as diagnostic markers of disease progression and stratification.

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“…LLOMe treatment results in the recruitment of LRRK2 and its substrate Rab8A to damaged endolysosomes (12, 13). We therefore tested LRRK2 and Rab8A positive vesicle formation in response to endolysosomal damage in M-CSF and GM-CSF differentiated BMDM.…”

Section: Resultsmentioning

confidence: 99%

LRRK2-dependent Rab GTPase phosphorylation in response to endolysosomal damage depends on macrophage differentiation

Herbst

Gutierrez

2020

Preprint

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The Parkinsons Disease (PD) kinase LRRK2 is highly expressed in immune cells such as macrophages. In these cells, LRRK2 regulates innate immune pathways and it is activated after membrane damage leading to the phosphorylation of the Rab GTPases Rab8A and Rab10. Due to their wide-range of functions in immunity and tissue remodelling, macrophages in vivo are phenotypically heterogeneous. In vitro systems are used to differentiate these cells into diverse macrophage subsets to mimic the populations observed in vivo. M-CSF and GM-CSF differentiated human blood monocytes are often used to generate monocyte-derived macrophages as a model for tissue macrophages. However, how LRRK2 is activated in different macrophage subsets after membrane damage is unknown. Here, we report that bone marrow derived macrophages and human monocyte-derived macrophages differentiated with either M-CSF or GM-CSF show different levels of LRRK2 activation after membrane damage. Notably, the membrane damaging agent LLOMe triggered LRRK2-dependent Rab8A and Rab10 phosphorylation primarily in GM-CSF differentiated macrophages. Moreover, LRRK2 and Rab8A were recruited to damaged endolysosomes in GM-CSF differentiated macrophages. Strikingly, GM-CSF differentiated macrophages recruited significantly more CHMP4B and Galectin-3 into damaged endolysosomes. These results suggest that LRRK2-regulated pathways of endolysosomal membrane damage and repair differ between macrophage subsets.

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LRRK2 mediates tubulation and vesicle sorting from membrane damaged lysosomes

Cited by 15 publications

References 56 publications

Pathogenic mutations in LRRK2 sequester Rab8a to damaged lysosomes and regulate transferrin-mediated iron uptake in microglia

Pathogenic mutations in LRRK2 sequester Rab8a to damaged lysosomes and regulate transferrin-mediated iron uptake in microglia

Divergent Effects of G2019S and R1441C LRRK2 Mutations on LRRK2 and Rab10 Phosphorylations in Mouse Tissues

LRRK2-dependent Rab GTPase phosphorylation in response to endolysosomal damage depends on macrophage differentiation

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