Kallikrein 6 signals through PAR1 and PAR2 to promote neuron injury and exacerbate glutamate neurotoxicity

Authors

  • Hyesook Yoon,

    1. Neurobiology of Disease Program, Mayo Medical and Graduate School, Rochester, Minnesota, USA
    2. Department of Physical Medicine and Rehabilitation, Mayo Medical and Graduate School, Rochester, Minnesota, USA
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  • Maja Radulovic,

    1. Neurobiology of Disease Program, Mayo Medical and Graduate School, Rochester, Minnesota, USA
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  • Jianmin Wu,

    1. Department of Physical Medicine and Rehabilitation, Mayo Medical and Graduate School, Rochester, Minnesota, USA
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  • Sachiko I. Blaber,

    1. Department of Biomedical Sciences, Florida State University College of Medicine, Tallahassee, Florida, USA
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  • Michael Blaber,

    1. Department of Biomedical Sciences, Florida State University College of Medicine, Tallahassee, Florida, USA
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  • Michael G. Fehlings,

    1. Department of Surgery, Toronto Western Research Institute, Toronto, Ontario, Canada
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  • Isobel A. Scarisbrick

    Corresponding author
    1. Neurobiology of Disease Program, Mayo Medical and Graduate School, Rochester, Minnesota, USA
    2. Department of Physical Medicine and Rehabilitation, Mayo Medical and Graduate School, Rochester, Minnesota, USA
    3. Department of Neurology, Mayo Medical and Graduate School, Rochester, Minnesota, USA
    • Address correspondence and reprint requests to Dr. Isobel A. Scarisbrick, Neurobiology of Disease Program, 642B Guggenheim Building, Mayo Clinic Rochester, 200 First St. SW., Rochester, MN 55905, USA. E-mail: scarisbrick.isobel@mayo.edu

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Abstract

CNS trauma generates a proteolytic imbalance contributing to secondary injury, including axonopathy and neuron degeneration. Kallikrein 6 (Klk6) is a serine protease implicated in neurodegeneration, and here we investigate the role of protease-activated receptors 1 (PAR1) and PAR2 in mediating these effects. First, we demonstrate Klk6 and the prototypical activator of PAR1, thrombin, as well as PAR1 and PAR2, are each elevated in murine experimental traumatic spinal cord injury (SCI) at acute or subacute time points. Recombinant Klk6 triggered extracellular signal-regulated kinase (ERK1/2) signaling in cerebellar granule neurons and in the NSC34 spinal cord motoneuron cell line, in a phosphoinositide 3-kinase and MEK-dependent fashion. Importantly, lipopeptide inhibitors of PAR1 or PAR2, and PAR1 genetic deletion, each reduced Klk6-ERK1/2 activation. In addition, Klk6 and thrombin promoted degeneration of cerebellar neurons and exacerbated glutamate neurotoxicity. Moreover, genetic deletion of PAR1 blocked thrombin-mediated cerebellar neurotoxicity and reduced the neurotoxic effects of Klk6. Klk6 also increased glutamate-mediated Bim signaling, poly-ADP-ribose polymerase cleavage and lactate dehydrogenase release in NSC34 motoneurons and these effects were blocked by PAR1 and PAR2 lipopeptide inhibitors. Taken together, these data point to a novel Klk6-signaling axis in CNS neurons that is mediated by PAR1 and PAR2 and is positioned to contribute to neurodegeneration.

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Here, we show kallikrein 6 (Klk6) and thrombin contribute to the proteolytic imbalance that occurs in CNS injury. Activation of the G-protein coupled receptor PAR1 was sufficient to mediate the neurotoxic effects of thrombin while KLK6 neurotoxicity involved activation of PAR1 and PAR2. In addition, both proteases exacerbated glutamate neurotoxicity. These data suggest Klk6, thrombin and PARs each represent new targets for the development of neuroprotective therapies.

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