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Chloroquine Restores Ganglioside Homeostasis and Improves Pathological and Behavioral Outcomes Post-stroke in the Rat

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Abstract

Perturbations of ganglioside homeostasis have been observed following stroke whereby toxic simple gangliosides GM2 and GM3 accumulate, while protective complex species GM1 and GD1 are reduced. Thus, there is a need for therapeutic interventions which can prevent ganglioside dysregulation after stroke. A pharmacological intervention using chloroquine was selected for its transient lysosomotropic properties which disrupt the activity of catabolic ganglioside enzymes. Chloroquine was administered both in vitro (0.1 μM), to primary cortical neurons exposed to GM3 toxicity, and in vivo (45 mg/kg i.p.), to 3-month-old male Wistar rats that underwent a severe stroke injury. Chloroquine was administered for seven consecutive days beginning 3 days prior to the stroke injury. Gangliosides were examined using MALDI imaging mass spectrometry at 3 and 21 days after the injury, and motor deficits were examined using the ladder task. Chloroquine treatment prevented ganglioside dysregulation 3 days post-stroke and partially prevented complex ganglioside depletion 21 days post-stroke. Exogenous GM3 was found to be toxic to primary cortical neurons which was protected by chloroquine treatment. Motor deficits were prevented in the forelimbs of stroke-injured rats with chloroquine treatment and was associated with decreased inflammation, neurodegeneration, and an increase in cell survival at the site of injury. Chloroquine administration prevents ganglioside dysregulation acutely, protects against GM3 toxicity in neurons, and is associated with long-term functional and pathological improvements after stroke in the rat. Therefore, targeting lipid dysregulation using lysosomotropic agents such as chloroquine may represent a novel therapeutic avenue for stroke injuries.

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Acknowledgements

We are grateful to Dilani Logan and Justin Kim for their assistance in the quantification of MALDI data and behavioral analyses.

Funding

This work was funded by NSERC, CIHR, CCNA operating grants and a CFI equipment grant to SNW, an NSERC operating grant to KK-CY, a CIHR emerging team grant to DFC and SNW, and an NSERC PGS-D award to SC.

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Authors and Affiliations

  1. Vulnerable Brain Laboratory, Department of Anatomy and Cell Biology, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, N6A 5C1, Canada

    Sarah Caughlin, Jeffrey Hepburn, Qingfan Liu, Lynn Wang, David F. Cechetto & Shawn N. Whitehead

  2. Department of Chemistry, Department of Biochemistry, Schulich School of Medicine and Dentistry, University of Western Ontario, London, ON, Canada

    Ken K.-C. Yeung

Authors
  1. Sarah Caughlin
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  2. Jeffrey Hepburn
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  3. Qingfan Liu
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  4. Lynn Wang
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  5. Ken K.-C. Yeung
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  7. Shawn N. Whitehead
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Contributions

SC conceptualized experiments, performed all MALDI-IMS, primary cortical neuron, and behavior experiments, analyzed the data, and wrote the manuscript. LW performed cell extraction and plating of all primary cortical neurons. JH performed and analyzed all immunohistochemistry experiments. DFC and KY conceptualized experiments and edited the manuscript. SNW conceptualized experiments and edited the manuscript and figures.

Corresponding author

Correspondence to Shawn N. Whitehead.

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The authors have no conflicts of interest to disclose.

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Caughlin, S., Hepburn, J., Liu, Q. et al. Chloroquine Restores Ganglioside Homeostasis and Improves Pathological and Behavioral Outcomes Post-stroke in the Rat. Mol Neurobiol 56, 3552–3562 (2019). https://doi.org/10.1007/s12035-018-1317-0

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