The activities of LRRK2 and GCase are positively correlated in clinical biospecimens and experimental models of Parkinson’s disease

Kedariti, Maria; Frattini, Emanuele; Baden, Pascale; Cogo, Susanna; Civiero, Laura; Ziviani, Elena; Aureli, Massimo; Kaganovich, Alice; Cookson, Mark; Stefanis, Leonidas; Surface, Matthew; Deleidi, Michela; Fonzo, Alessio Di; Alcalay, Roy N.; Rideout, Hardy J.; Greggio, Elisa; Plotegher, Nicoletta

doi:10.1101/2021.09.27.461935

Cited by 4 publications

(3 citation statements)

References 48 publications

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“…As the G2019S mutation causes an increase in LRRK2 kinase activity, the prediction from these studies would be that inhibition of LRRK2 kinase activity would result in an increase in GBA protein levels and a decrease in specific activity. In a recent work, Kedariti et al showed that pharmacological inhibition of G2019S LRRK2 in HEK293T cells caused a significant reduction in GCase activity when it was normalized by GBA levels, being consistent with the results in our study [16].…”

Section: Discussionsupporting

confidence: 93%

Seventy-Two-Hour LRRK2 Kinase Activity Inhibition Increases Lysosomal GBA Expression in H4, a Human Neuroglioma Cell Line

Ruz

Alcantud

Vives

et al. 2022

IJMS

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Mutations in LRRK2 and GBA1 are key contributors to genetic risk of developing Parkinson’s disease (PD). To investigate how LRRK2 kinase activity interacts with GBA and contributes to lysosomal dysfunctions associated with the pathology of PD. The activity of the lysosomal enzyme β-Glucocerebrosidase (GCase) was assessed in a human neuroglioma cell model treated with two selective inhibitors of LRKK2 kinase activity (LRRK2-in-1 and MLi-2) and a GCase irreversible inhibitor, condutirol-beta-epoxide (CBE), under 24 and 72 h experimental conditions. We observed levels of GCase activity comparable to controls in response to 24 and 72 h treatments with LRRK2-in-1 and MLi-2. However, GBA protein levels increased upon 72 h treatment with LRRK2-in-1. Moreover, LC3-II protein levels were increased after both 24 and 72 h treatments with LRRK2-in-1, suggesting an activation of the autophagic pathway. These results highlight a possible regulation of lysosomal function through the LRRK2 kinase domain and suggest an interplay between LRRK2 kinase activity and GBA. Although further investigations are needed, the enhancement of GCase activity might restore the defective protein metabolism seen in PD.

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Section: Discussionsupporting

confidence: 93%

Seventy-Two-Hour LRRK2 Kinase Activity Inhibition Increases Lysosomal GBA Expression in H4, a Human Neuroglioma Cell Line

Ruz

Alcantud

Vives

et al. 2022

IJMS

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“…All participants provided written informed consent to take part in the study, and the study protocol was approved by the IRBs of both Columbia Univers0ity (CUIMC). PBMCs were isolated using standard protocols as previously described (83). Briefly, BD Vacutainer tubes (Sodium Heparin or Sodium Citrate) were used to collect whole blood from study participants.…”

Section: Peripheral Blood Mononuclear Cell (Pbmc) Isolation and Trans...mentioning

confidence: 99%

A potential patient stratification biomarker for Parkinso’s disease based on LRRK2 kinase-mediated centrosomal alterations in peripheral blood-derived cells

Naaldijk

Fernández

Fasiczka

et al. 2023

Preprint

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Parkinson's disease (PD) is a common neurodegenerative movement disorder and leucine-rich repeat kinase 2 (LRRK2) is a promising therapeutic target for disease intervention. However, the ability to stratify patients who will benefit from such treatment modalities based on shared etiology is critical for the success of disease-modifying therapies. Ciliary and centrosomal alterations are commonly associated with pathogenic LRRK2 kinase activity and can be detected in many cell types. We previously found centrosomal deficits in immortalized lymphocytes from G2019S-LRRK2 PD patients. Here, to investigate whether such deficits may serve as a potential blood biomarker for PD which is susceptible to LRKK2 inhibitor treatment, we characterized patient-derived cells from distinct PD cohorts. We report centrosomal alterations in peripheral cells from a subset of early-stage idiopathic PD patients which is mitigated by LRRK2 kinase inhibition, supporting a role for aberrant LRRK2 activity in idiopathic PD. Centrosomal defects are detected in R1441G-LRRK2 and G2019S-LRRK2 PD patients and in non-manifesting LRRK2 mutation carriers, indicating that they acumulate prior to a clinical PD diagnosis. They are present in immortalized cells as well as in primary lymphocytes from peripheral blood. These findings indicate that analysis of centrosomal defects as a blood-based patient stratification biomarker may help nominate PD patients who will benefit from LRRK2-related therapeutics.

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“…hESCs are pluripotent cells that have been derived from the inner cell mass of an early-stage embryo (Thomson et al, 1998) while iPSCs are generated from adult somatic cells, usually skin fibroblast cells, that are induced to pluripotency via a combination of four transcription factors (Takahashi and Yamanaka, 2006;Takahashi et al, 2007). Advances over the last decade or so, in defining differentiation conditions have made it possible to directly drive differentiation of human pluripotent cells to DA neurons and use these derived neurons to model PD in a dish (Yang et al, 2008;Chambers et al, 2009;Swistowski et al, 2010;Kriks et al, 2011;Gonzalez et al, 2013;Zhang et al, 2014;Singh et al, 2017;Barbuti et al, 2020;Kedariti et al, 2022;Stern et al, 2022). Multiple studies from different research laboratories have also shown that these derived neurons not only have cellular and biochemical characteristics very similar to the neurons developed in the human brain but also are functional when transplanted into animal models and thus, may have the potential to contribute to regenerative treatment options for the disease (Byers et al, 2011;Kriks et al, 2011;Doi et al, 2014;Grealish et al, 2014;Byers et al, 2015;Hoban et al, 2020;Behl et al, 2022).…”

Section: Introductionmentioning

confidence: 99%

Identification of DOT1L inhibitor in a screen for factors that promote dopaminergic neuron survival

Cui

Carey

Pera

2022

Front. Aging Neurosci.

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Parkinson’s disease (PD) is a common neurodegenerative disorder characterized by the progressive loss of dopaminergic (DA) neurons in the substantia nigra region of the midbrain. Diagnostic criteria for PD require that at least two of three motor signs are observed: tremor, rigidity, and/or bradykinesia. The most common and effective treatment for PD is Levodopa (L-DOPA) which is readily converted to DA and has been the primary treatment since the 1960’s. Dopamine agonists have also been developed but are less effective than L-DOPA. Although the lack of a model system to study PD has hampered efforts to identify treatments, diverse screening strategies have been proposed for identification of new pharmaceutical candidates. Here, we describe a pilot screen to identify candidate molecules from a bioactive compound library, that might increase formation, maintenance and/or survival of DA neurons in vitro. The screen used a previously characterized reporter construct consisting of the luciferase gene inserted downstream of the endogenous tyrosine hydroxylase (TH) gene and neurons differentiated from human pluripotent stem cells for 18 days. The reporter mimics expression of TH and includes a secreted luciferase whose activity can be measured non-invasively over multiple timepoints. Screening of the bioactive compound library resulted in the identification of a single molecule, SGC0946, that is an inhibitor of DOT1L (Disruptor Of Telomeric silencing 1-Like) which encodes a widely-conserved histone H3K79 methyltransferase that is able to both activate and repress gene transcription. Our results indicate that SGC0946 increased reporter luciferase activity with a single treatment for 48-h post-plating being equivalent to continuous treatment. Moreover, data suggested that the total number of neurons differentiated in the assays was comparable from experiment to experiment under different SGC0946 treatments over time. In contrast, data suggested that the survival and/or maintenance of DA neurons might be specifically enhanced by SGC0946 treatment. These results document the feasibility of a set of tools for further exploration of small molecules that may impact DA neuron differentiation, maintenance and/or survival. Results provide evidence in support of other reports that indicate inhibition of DOT1L may play an important role in maintenance and survival of neural progenitor cells (NPCs) and their lineage-specific differentiation.

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The activities of LRRK2 and GCase are positively correlated in clinical biospecimens and experimental models of Parkinson’s disease

Cited by 4 publications

References 48 publications

Seventy-Two-Hour LRRK2 Kinase Activity Inhibition Increases Lysosomal GBA Expression in H4, a Human Neuroglioma Cell Line

Seventy-Two-Hour LRRK2 Kinase Activity Inhibition Increases Lysosomal GBA Expression in H4, a Human Neuroglioma Cell Line

A potential patient stratification biomarker for Parkinso’s disease based on LRRK2 kinase-mediated centrosomal alterations in peripheral blood-derived cells

Identification of DOT1L inhibitor in a screen for factors that promote dopaminergic neuron survival

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