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Leucine-rich repeat kinase 2 and alternative splicing in Parkinson's disease§

Authors

  • David A. Elliott PhD,

    1. Neuroscience Research Australia, Sydney, New South Wales, Australia
    2. School of Medical Sciences, University of New South Wales, Sydney, New South Wales, Australia
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  • Woojin S. Kim PhD,

    1. Neuroscience Research Australia, Sydney, New South Wales, Australia
    2. School of Medical Sciences, University of New South Wales, Sydney, New South Wales, Australia
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  • Sarsha Gorissen BSc (Hons),

    1. Neuroscience Research Australia, Sydney, New South Wales, Australia
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  • Glenda M. Halliday PhD,

    1. Neuroscience Research Australia, Sydney, New South Wales, Australia
    2. School of Medical Sciences, University of New South Wales, Sydney, New South Wales, Australia
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  • John B.J. Kwok PhD

    Corresponding author
    1. Neuroscience Research Australia, Sydney, New South Wales, Australia
    2. School of Medical Sciences, University of New South Wales, Sydney, New South Wales, Australia
    • Neuroscience Research Australia, Barker Street, Randwick, Sydney, New South Wales, 2031 Australia
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  • Funding agencies: The authors thank Dr. I.P. Blair (ANZAC Institute, Sydney, New South Wales, Australia) for the gift of the mutant, TARDBP, and FUS mutant complementary DNAs (cDNAs), and Prof. W.P. Gai (Flinders University, Adelaide, South Australia, Australia) for the gift of the wild-type and mutant LRRK2 cDNAs. Brain tissue was received from the Sydney Brain Bank at Neuroscience Research Australia, which is supported by Neuroscience Research Australia, the University of New South Wales, and the National Health and Medical Research Council (NHMRC) of Australia, and the New South Wales Tissue Resource Center at the University of Sydney, which is supported by the NHMRC, the Schizophrenia Research Institute, and the National Institute of Alcohol Abuse and Alcoholism (National Institutes of Health; R24AA012725). This study was funded by the NHMRC (project grant 510218, to J.B.J.K.; fellowship 630434, to G.M.H.). D.E. was supported by a Viertel Post-Doctoral Fellowship awarded by Alzheimer's Australia Research.

  • Relevant conflicts of interest/financial disclosures: Nothing to report.

  • §

    Full financial disclosures and conflicts of interest may be found in the online version of this article.

Abstract

Mutations of the leucine-rich repeat kinase 2 (LRRK2) gene are the most common genetic cause of Parkinson's disease (PD) and are associated with pleiomorphic neuropathology. We hypothesize that LRRK2 mediates its pathogenic effect through alternative splicing of neurodegeneration genes. Methods used in this study included western blotting analysis of subcellular protein fractions, exon-array analysis of RNA from cultured neuroblastoma cells transfected with LRRK2 expression vectors, and reverse-transcription polymerase chain reaction (RT-PCR) of RNA from cultured cells and postmortem tissue. Overexpression of the LRRK2 G2019S mutant resulted in a significant (2.6-fold; P = 0.020) decrease in nuclear transactive response DNA-binding protein 43 levels. Exon-array analyses revealed that wild-type LRRK2 had a significant effect on the expression of genes with nuclear (P < 10−22) and cell-cycle functions (P < 10−15). We replicated changes in gene expression in 30% of selected genes by quantitative RT-PCR. Overexpression of LRRK2 resulted in the altered splicing of two genes associated with PD, with an increased inclusion of exon 10 of microtubule-associated protein tau (1.7-fold; P = 0.001) and exon 5 of the alpha-synuclein (SNCA) gene (1.6-fold; P =0.005). Moreover, overexpression of LRRK2 (G2019S) and two mutant genes associated with neurodegeneration, TARDBP (M337V) and FUS (R521H), were associated with decreased inclusion out of the dystonin (DST) 1e precursor exons in SK-N-MC cells. Altered splicing of SNCA (1.9-fold; P < 0.001) and DST genes (log2 2.3-fold; P = 0.005) was observed in a cohort of PD, compared with neurologically healthy, brains. This suggests that aberrant RNA metabolism is an important contributor to idiopathic PD. © 2012 Movement Disorder Society

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