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Quinacrine and Niclosamide Promote Neurite Growth in Midbrain Dopaminergic Neurons Through the Canonical BMP-Smad Pathway and Protect Against Neurotoxin and α-Synuclein-Induced Neurodegeneration

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Abstract

Parkinson’s disease is a neurodegenerative disorder characterised by nigrostriatal dopaminergic degeneration, and intracellular α-synuclein aggregation. Current pharmacological treatments are solely symptomatic so there is a need to identify agents that can slow or stop dopaminergic degeneration. One proposed class of therapeutics are neurotrophic factors which promote the survival of nigrostriatal dopaminergic neurons. However, neurotrophic factors need to be delivered directly to the brain. An alternative approach may be to identify pharmacological agents which can reach the brain to stimulate neurotrophic factor expression and/or their signalling pathways in dopaminergic neurons. BMP2 is a neurotrophic factor that is expressed in the human substantia nigra; exogenous BMP2 administration protects against dopaminergic degeneration in in vitro models of PD. In this study, we investigated the neurotrophic potential of two FDA-approved drugs, quinacrine and niclosamide, that are modulators of BMP2 signalling. We report that quinacrine and niclosamide, like BMP2, significantly increased neurite length, as a readout of neurotrophic action, in SH-SY5Y cells and dopaminergic neurons in primary cultures of rat ventral mesencephalon. We also show that these effects of quinacrine and niclosamide require the activation of BMP-Smad signalling. Finally, we demonstrate that quinacrine and niclosamide are neuroprotective against degeneration induced by the neurotoxins, MPP+ and 6-OHDA, and by viral-mediated overexpression of α-synuclein in vitro. Collectively, this study identifies two drugs, that are safe for use in patients' to 'are approved for human use, that exert neurotrophic effects on dopaminergic neurons through modulation of BMP-Smad signalling. This rationalises the further study of drugs that target the BMP-Smad pathway as potential neuroprotective pharmacotherapy for Parkinson’s disease.

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Data Availability

The data that support the findings of this study are available from the corresponding author, upon reasonable request.

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Funding

This publication has emanated from research conducted with the financial support of a RISAM PhD scholarship from Munster Technological University (R00094948) and a research grant from Science Foundation Ireland (SFI) under the grant numbers 15/CDA/3498 (G.O’K.).

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Authors and Affiliations

  1. Department of Biological Sciences, Munster Technological University, Cork, Ireland

    Susan R. Goulding

  2. Department of Anatomy and Neuroscience and Cork Neuroscience Centre, University College Cork, Cork, Ireland

    Susan R. Goulding, Aideen M. Sullivan, Louise M. Collins & Gerard W. O’Keeffe

  3. Department of Psychiatry and Neurosciences, Cervo Brain Research Centre, Université Laval, Quebec, QC, Canada

    Martin Lévesque

  4. APC Microbiome Institute, University College Cork, Cork, Ireland

    Aideen M. Sullivan & Gerard W. O’Keeffe

  5. Department of Physiology, University College Cork, Cork, Ireland

    Louise M. Collins

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  1. Susan R. Goulding
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  2. Martin Lévesque
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  4. Louise M. Collins
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  5. Gerard W. O’Keeffe
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Contributions

SG performed the experiments, analysed the data and co-wrote the manuscript. ML, AS, LC and GOK co-wrote the manuscript. SG, LC and GOK designed the experiments. LC and GOK supervised the study.

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Correspondence to Louise M. Collins or Gerard W. O’Keeffe.

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ESM 1

Fig. 1. SH-SY5Y cells as a tool to study drugs affecting BMP-Smad signalling. Representative photomicrographs showing immunocytochemical staining for (a) BMPR1B, (b) BMPR2 and (c) Smad1/5/8 expression in SH-SY5Y cells. (d) Representative photomicrographs of immunocytochemical staining of phospho-Smad1/5/8 and (e) intensity of phospho-Smad1/5/8 as measured using ELISA in SH-SY5Y cells after treatment with 50 ng/ml rhBMP2 with and without 1 μg/ml dorsomorphin for 2 h. (f) Total neurite length and (h) representative photomicrographs of SH-SY5Y cells after treatment with 10, 50 or 200 ng/ml rhBMP2 for 24 h. (g) Total neurite length and (i) representative photomicrographs of SH-SY5Y cells after treatment with 1 μg/ml dorsomorphin with and without 50 ng/ml rhBMP2 for 24 h. All data are presented as mean ± SEM from at least three experiments. (*p < 0.05, **p < 0.01, ***p < 0.001 vs. control; +++p < 0.001 vs rhBMP2. One-way ANOVA with Tukey’s post-hoc test). Fig. 2. AAV-α-synuclein affects neurite length of cultured dopaminergic neurons in a concentration-dependent manner. (a) Graph and (b) representative photomicrographs of TH+ neurons transduced with AAV2/6-GFP or AAV2/6-α-synuclein at the MOI’s indicated for 5 DIV. Scale bar = 50 μm. All data are presented as mean ± SEM from at least three experiments. (**p < 0.01 vs. control; Unpaired Student’s t test). (DOCX 528 kb)

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Goulding, S.R., Lévesque, M., Sullivan, A.M. et al. Quinacrine and Niclosamide Promote Neurite Growth in Midbrain Dopaminergic Neurons Through the Canonical BMP-Smad Pathway and Protect Against Neurotoxin and α-Synuclein-Induced Neurodegeneration. Mol Neurobiol 58, 3405–3416 (2021). https://doi.org/10.1007/s12035-021-02351-8

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