Optimisation of LRRK2 inhibitors and assessment of functional efficacy in cell-based models of neuroinflammation

Munoz, Lenka; Kavanagh, Madeline E.; Phoa, Athena; Heng, Benjamin; Dzamko, Nicolas; Chen, Ew-Jun; Doddareddy, Munikumar Reddy; Guillemin, Gilles J.; Kassiou, Michael

doi:10.1016/j.ejmech.2015.03.003

Cited by 30 publications

(20 citation statements)

References 25 publications

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“…Inflammation and oxidative stress are closely associated with the pathogenesis of many neurodegenerative disorders including AD, PD and multiple sclerosis [65]. Numerous studies have reported that LRRK2 is involved in neuroinflammation and inflammatory signaling [66, 67]. Microglial LRRK2 is linked to the inflammatory response associated with PD [68, 69].…”

Section: Discussionmentioning

confidence: 99%

LRRK2 kinase plays a critical role in manganese-induced inflammation and apoptosis in microglia

et al. 2019

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Long-term exposure to elevated levels of manganese (Mn) causes manganism, a neurodegenerative disorder with Parkinson’s disease (PD)-like symptoms. Increasing evidence suggests that leucine-rich repeat kinase 2 (LRRK2), which is highly expressed in microglia and macrophages, contributes to the inflammation and neurotoxicity seen in autosomal dominant and sporadic PD. As gene-environment interactions have emerged as important modulators of PD-associated toxicity, LRRK2 may also mediate Mn-induced inflammation and pathogenesis. In this study, we investigated the role of LRRK2 in Mn-induced toxicity using human microglial cells (HMC3), LRRK2-wild-type (WT) and LRRK2-knockout (KO) RAW264.7 macrophage cells. Results showed that Mn activated LRRK2 kinase by phosphorylation of its serine residue at the 1292 position (S1292) as a marker of its kinase activity in macrophage and microglia, while inhibition with GSK2578215A (GSK) and MLi-2 abolished Mn-induced LRRK2 activation. LRRK2 deletion and its pharmacological inhibition attenuated Mn-induced apoptosis in macrophages and microglia, along with concomitant decreases in the pro-apoptotic Bcl-2-associated X (Bax) protein. LRRK2 deletion also attenuated Mn-induced production of reactive oxygen species (ROS) and the pro-inflammatory cytokine TNF-α. Mn-induced phosphorylation of mitogen-activated protein kinase (MAPK) p38 and ERK signaling proteins was significantly attenuated in LRRK2 KO cells and GSK-treated cells. Moreover, inhibition of MAPK p38 and ERK as well as LRRK2 attenuated Mn-induced oxidative stress and cytotoxicity. These findings suggest that LRRK2 kinase activity plays a critical role in Mn-induced toxicity via downstream activation of MAPK signaling in macrophage and microglia. Collectively, these results suggest that LRRK2 could be a potential molecular target for developing therapeutics to treat Mn-related neurodegenerative disorders.

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Section: Discussionmentioning

confidence: 99%

LRRK2 kinase plays a critical role in manganese-induced inflammation and apoptosis in microglia

et al. 2019

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“…Following siRNA treatment, which efficiently silenced LRRK2 expression (Figure S1B), IMG cells were transfected with the Lamp1.GFP plasmid to label the lysosomes. Consistent with the adult fly brains, lysosomes of IMG cells treated with the LRRK2 siRNA or inhibitor LRRK2-IN-1 (Deng et al, 2011;Munoz et al, 2015) were enlarged and fewer in number (Figures 1E-1H). Furthermore, Transmission Electron Microscopy (TEM) analysis also revealed that lysosomes were enlarged with abnormal morphology and fewer in number upon glial dLRRK depletion in adult flies of .…”

Section: Reducing Dlrrk Expression In Glia Causes Enlarged Lysosomes Fewer In Numbersupporting

confidence: 75%

A LRRK2/dLRRK-mediated lysosomal pathway that contributes to glial cell death and DA neuron survival

Wang

2021

Preprint

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Mutations in leucine-rich repeat kinase 2 (LRRK2) are the most common cause of familial and sporadic Parkinson’s disease (PD). A plethora of evidence has indicated a role for LRRK2 in endolysosomal trafficking in neurons, while LRRK2 function in glia, although highly expressed, remains largely unknown. Here we present evidence that LRRK2/dLRRK mediates a glial lysosomal pathway that contributes to the mechanism of PD. Independent of its kinase activity, glial LRRK2/dLRRK knockdown in the immortalized microglial cells or flies results in enlarged and swelling lysosomes fewer in number. These lysosomes are less mobile, wrongly acidified, and exhibit defective membrane permeability and reduced activity of the lysosome hydrolase cathespin B. In addition, microglial LRRK2 depletion causes increased Caspase 3 levels, leading to glial apoptosis, dopaminergic neurodegeneration, and locomotor deficits in an age-dependent manner. Taken together, these findings demonstrate a functional role of LRRK2/dLRRK in regulating the glial lysosomal pathway; deficits in lysosomal biogenesis and function linking to glial apoptosis potentially underlie the mechanism of DA neurodegeneration, contributing to the progression of PD.

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“…Thus, it is tempting to speculate that LRRK2 activity contributes to sustainment of neuroinflammation and that pharmacological inhibition treatments may be effective at attenuating chronic neuroinflammation and neurodegeneration in PD patients. In this context, several groups intensely worked to identify LRRK2 inhibitors with good physiochemical and pharmacokinetic properties, selectivity and blood-brain barrier permeability, against both kinase [ 60 – 63 ] and GTPase activities [ 64 ]. While LRRK2 inhibition has been proven to be effective at reducing pathological LRRK2 phenotypes [ 65 ], other studies showed that Lrrk2 knock-out mice or non-human primates treated with chronic doses of LRRK2 inhibitors exhibit side effects and morphologic changes in peripheral tissue [ 66 , 67 ].…”

Section: Discussionmentioning

confidence: 99%

Leucine-rich repeat kinase 2 positively regulates inflammation and down-regulates NF-κB p50 signaling in cultured microglia cells

Russo

Berti

Plotegher

et al. 2015

J Neuroinflammation

111

114

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BackgroundOver-activated microglia and chronic neuroinflammation contribute to dopaminergic neuron degeneration and progression of Parkinson’s disease (PD). Leucine-rich repeat kinase 2 (LRRK2), a kinase mutated in autosomal dominantly inherited and sporadic PD cases, is highly expressed in immune cells, in which it regulates inflammation through a yet unclear mechanism.MethodsHere, using pharmacological inhibition and cultured Lrrk2−/− primary microglia cells, we validated LRRK2 as a positive modulator of inflammation and we investigated its specific function in microglia cells.ResultsInhibition or genetic deletion of LRRK2 causes reduction of interleukin-1β and cyclooxygenase-2 expression upon lipopolysaccharide-mediated inflammation. LRRK2 also takes part of the signaling trigged by α-synuclein fibrils, which culminates in induction of inflammatory mediators. At the molecular level, loss of LRRK2 or inhibition of its kinase activity results in increased phosphorylation of nuclear factor kappa-B (NF-κB) inhibitory subunit p50 at S337, a protein kinase A (PKA)-specific phosphorylation site, with consequent accumulation of p50 in the nucleus.ConclusionsTaken together, these findings point to a role of LRRK2 in microglia activation and sustainment of neuroinflammation and in controlling of NF-κB p50 inhibitory signaling. Understanding the molecular pathways coordinated by LRRK2 in activated microglia cells after pathological stimuli such us fibrillar α-synuclein holds the potential to provide novel targets for PD therapeutics.

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Optimisation of LRRK2 inhibitors and assessment of functional efficacy in cell-based models of neuroinflammation

Cited by 30 publications

References 25 publications

LRRK2 kinase plays a critical role in manganese-induced inflammation and apoptosis in microglia

LRRK2 kinase plays a critical role in manganese-induced inflammation and apoptosis in microglia

A LRRK2/dLRRK-mediated lysosomal pathway that contributes to glial cell death and DA neuron survival

Leucine-rich repeat kinase 2 positively regulates inflammation and down-regulates NF-κB p50 signaling in cultured microglia cells

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