Faculty Opinions recommendation of The Parkinson's disease VPS35[D620N] mutation enhances LRRK2-mediated Rab protein phosphorylation in mouse and human.

Mutations that activate the LRRK2 (leucine-rich repeat protein kinase 2) protein kinase predispose to Parkinson's disease, suggesting that LRRK2 inhibitors might have therapeutic benefit. Recent work has revealed that LRRK2 phosphorylates a subgroup of 14 Rab proteins, including Rab10, at a specific residue located at the centre of its effector-binding switch-II motif. In the present study, we analyse the selectivity and sensitivity of polyclonal and monoclonal phospho-specific antibodies raised against nine different LRRK2-phosphorylated Rab proteins (Rab3A/3B/3C/3D, Rab5A/5B/5C, Rab8A/8B, Rab10, Rab12, Rab29[T71], Rab29[S72], Rab35 and Rab43). We identify rabbit monoclonal phospho-specific antibodies (MJFF-pRAB10) that are exquisitely selective for LRRK2-phosphorylated Rab10, detecting endogenous phosphorylated Rab10 in all analysed cell lines and tissues, including human brain cingulate cortex. We demonstrate that the MJFF-pRAB10 antibodies can be deployed to assess enhanced Rab10 phosphorylation resulting from pathogenic (R1441C/G or G2019S) LRRK2 knock-in mutations as well as the impact of LRRK2 inhibitor treatment. We also identify rabbit monoclonal antibodies displaying broad specificity (MJFF-pRAB8) that can be utilised to assess LRRK2-controlled phosphorylation of a range of endogenous Rab proteins, including Rab8A, Rab10 and Rab35. The antibodies described in the present study will help with the assessment of LRRK2 activity and examination of which Rab proteins are phosphorylated in vivo. These antibodies could also be used to assess the impact of LRRK2 inhibitors in future clinical trials.

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Identification of Potential miRNA-mRNA Regulatory Network Contributing to Parkinson’s Disease

10.21203/rs.3.rs-779087/v1 ◽

2021 ◽

Author(s):

Xi Yin ◽

Miao Wang ◽

Wei Wang ◽

Tong Chen ◽

Ge Song ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Regulatory Network ◽

Enrichment Analysis ◽

Functional Enrichment ◽

Pathway Enrichment Analysis ◽

Hub Genes ◽

Rab Protein ◽

Healthy Donors ◽

Geo Database

Abstract Parkinson’s disease (PD) is a common neurodegenerative disease and the mechanism underlying PD pathogenesis is incompletely understood. Increasing evidence indicates that microRNA (miRNA) plays critical regulatory role in the pathogenesis of PD. This study aimed to determine the miRNA-mRNA regulatory network for PD. The differentially expressed miRNAs (DEmis) and genes (DEGs) between PD patients and healthy donors were screened from miRNA dataset GSE16658 and mRNA dataset GSE100054 downloaded from the Gene Expression Omnibus (GEO) database. Target genes of the DEmis were selected when predicted by 3 or 4 online databases and overlapped with DEGs from GSE100054. Next, Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment analysis was conducted by Database for Annotation, Visualization and Integrated Discovery (DAVID) and Metascape analytic tool. The correlation between the screened genes and PD was evaluated by the online tool Comparative Toxicogenomics Database (CTD). The protein-protein interactions (PPI) network was built by STRING platform. Finally, we testify the expression of members of the miRNA-mRNA regulatory network in the blood samples collected from PD patients and healthy donors by using qRT-PCR. 1505 upregulated and 1302 downregulated DEGs, 77 upregulated DEmis and 112 downregulated DEmis were preliminarily screened from GEO database. Through further functional enrichment analysis, 10 PD-related hub genes were selected, including RAC1, IRS2, LEPR, PPARGC1A, CAMKK2, RAB10, RAB13, RAB27B, RAB11A and JAK2, which were mainly involved in Rab protein signaling transduction, AMPK signaling pathway and signaling by Leptin. The miRNA-mRNA regulatory network was constructed with 10 hub genes and their interacting miRNAs overlapped with DEmis, including miR-30e-5p, miR-142-3p, miR-101-3p, miR-32-3p, miR-508-5p, miR-642a-5p, miR-19a-3p and miR-21-5p. Analysis on clinical samples verified significant upregulation of LEPR and downregulation of miR-101-3p in PD patients compared with healthy donors. In the study, the potential miRNA-mRNA regulatory network was constructed in PD, which may provide novel insight into pathogenesis and treatment of PD.

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The Parkinson's disease VPS35[D620N] mutation enhances LRRK2-mediated Rab protein phosphorylation in mouse and human

Biochemical Journal ◽

10.1042/bcj20180248 ◽

2018 ◽

Vol 475 (11) ◽

pp. 1861-1883 ◽

Cited By ~ 57

Author(s):

Rafeeq Mir ◽

Francesca Tonelli ◽

Pawel Lis ◽

Thomas Macartney ◽

Nicole K. Polinski ◽

...

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Rab Proteins ◽

Missense Mutations ◽

Knock Out ◽

Rab Protein ◽

Retromer Complex ◽

Pathogenic Mutations ◽

Healthy Donors ◽

Mouse Tissues

Missense mutations in the LRRK2 (Leucine-rich repeat protein kinase-2) and VPS35 genes result in autosomal dominant Parkinson's disease. The VPS35 gene encodes for the cargo-binding component of the retromer complex, while LRRK2 modulates vesicular trafficking by phosphorylating a subgroup of Rab proteins. Pathogenic mutations in LRRK2 increase its kinase activity. It is not known how the only thus far described pathogenic VPS35 mutation, [p.D620N] exerts its effects. We reveal that the VPS35[D620N] knock-in mutation strikingly elevates LRRK2-mediated phosphorylation of Rab8A, Rab10, and Rab12 in mouse embryonic fibroblasts. The VPS35[D620N] mutation also increases Rab10 phosphorylation in mouse tissues (the lung, kidney, spleen, and brain). Furthermore, LRRK2-mediated Rab10 phosphorylation is increased in neutrophils as well as monocytes isolated from three Parkinson's patients with a heterozygous VPS35[D620N] mutation compared with healthy donors and idiopathic Parkinson's patients. LRRK2-mediated Rab10 phosphorylation is significantly suppressed by knock-out or knock-down of VPS35 in wild-type, LRRK2[R1441C], or VPS35[D620N] cells. Finally, VPS35[D620N] mutation promotes Rab10 phosphorylation more potently than LRRK2 pathogenic mutations. Available data suggest that Parkinson's patients with VPS35[D620N] develop the disease at a younger age than those with LRRK2 mutations. Our observations indicate that VPS35 controls LRRK2 activity and that the VPS35[D620N] mutation results in a gain of function, potentially causing PD through hyperactivation of the LRRK2 kinase. Our findings suggest that it may be possible to elaborate compounds that target the retromer complex to suppress LRRK2 activity. Moreover, patients with VPS35[D620N] associated Parkinson's might benefit from LRRK2 inhibitor treatment that have entered clinical trials in humans.

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Quantitative phosphoproteomic profiling of PINK1-deficient cells identifies phosphorylation changes in nuclear proteins

Molecular BioSystems ◽

10.1039/c3mb70565j ◽

2014 ◽

Vol 10 (7) ◽

pp. 1719-1729 ◽

Cited By ~ 6

Author(s):

Xiaoyan Qin ◽

Chaoya Zheng ◽

John R. Yates III ◽

Lujian Liao

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Protein Phosphorylation ◽

Nuclear Proteins

A quantitative phosphoproteomic method coupled with siRNA mediated silencing of a kinase associated with Parkinson's disease was applied to measure phosphorylation changes. The results indicate alterations in protein phosphorylation downstream of this kinase, potentially expanding our understanding of its function.

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Rab GTPases in Parkinson’s disease: a primer

Essays in Biochemistry ◽

10.1042/ebc20210016 ◽

2021 ◽

Author(s):

Antonio Jesús Lara Ordóñez ◽

Rachel Fasiczka ◽

Yahaira Naaldijk ◽

Sabine Hilfiker

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Membrane Trafficking ◽

Protein Function ◽

Current Knowledge ◽

Rab Gtpases ◽

Intracellular Membrane ◽

Master Regulators ◽

Rab Protein ◽

Physiological Properties

Abstract Parkinson’s disease is a prominent and debilitating movement disorder characterized by the death of vulnerable neurons which share a set of structural and physiological properties. Over the recent years, increasing evidence indicates that Rab GTPases can directly as well as indirectly contribute to the cellular alterations leading to PD. Rab GTPases are master regulators of intracellular membrane trafficking events, and alterations in certain membrane trafficking steps can be particularly disruptive to vulnerable neurons. Here, we describe current knowledge on the direct links between altered Rab protein function and PD pathomechanisms.

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Effects of repetitive transcranial magnetic stimulation on voice and speech in Parkinson's disease

The Journal of Laryngology & Otology ◽

10.1017/s0022215106002787 ◽

2005 ◽

pp. 1

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Transcranial Magnetic Stimulation ◽

Magnetic Stimulation ◽

Repetitive Transcranial Magnetic Stimulation ◽

Voice And Speech

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Emerging Drugs and Targets for Parkinson's Disease

10.1039/9781849737357 ◽

2013 ◽

Cited By ~ 3

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease

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Evaluation of Efferent Auditory System and Hearing Quality in Parkinson's Disease: Is the Difficulty in Speech Understanding in Complex Listening Conditions Related to Neural Degeneration or Aging?

Journal of Speech Language and Hearing Research ◽

10.1044/2020_jslhr-20-00337 ◽

2020 ◽

pp. 1-9

Author(s):

Nuriye Yıldırım Gökay ◽

Bülent Gündüz ◽

Fatih Söke ◽

Recep Karamert

Keyword(s):

Parkinson’S Disease ◽

Parkinson's Disease ◽

Auditory System ◽

Otoacoustic Emissions ◽

Pure Tone ◽

Pure Tone Audiometry ◽

Auditory Pathway ◽

Normal Hearing ◽

System Function ◽

Distortion Product

Purpose The effects of neurological diseases on the auditory system have been a notable issue for investigators because the auditory pathway is closely associated with neural systems. The purposes of this study are to evaluate the efferent auditory system function and hearing quality in Parkinson's disease (PD) and to compare the findings with age-matched individuals without PD to present a perspective on aging. Method The study included 35 individuals with PD (mean age of 48.50 ± 8.00 years) and 35 normal-hearing peers (mean age of 49 ± 10 years). The following tests were administered for all participants: the first section of the Speech, Spatial and Qualities of Hearing Scale; pure-tone audiometry, speech audiometry, tympanometry, and acoustic reflexes; and distortion product otoacoustic emissions (DPOAEs) and contralateral suppression of DPOAEs. SPSS Version 25 was used for statistical analyses, and values of p < .05 were considered statistically significant. Results There were no statistically significant differences in the pure-tone audiometry thresholds and DPOAE responses between the individuals with PD and their normal-hearing peers ( p = .732). However, statistically significant differences were found between the groups in suppression levels of DPOAEs and hearing quality ( p < .05). In addition, a statistically significant and positive correlation was found between the amount of suppression at some frequencies and the Speech, Spatial and Qualities of Hearing Scale scores. Conclusions This study indicates that medial olivocochlear efferent system function and the hearing quality of individuals with PD were affected adversely due to the results of PD pathophysiology on the hearing system. For optimal intervention and follow-up, tasks related to hearing quality in daily life can also be added to therapies for PD.

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