Complement protein C1q is a therapeutic target for neuropathic pain

Activation of spinal microglia following peripheral nerve injury is a central component of neuropathic pain pathology. While the contributions of microglia-mediated immune and neurotrophic signalling have been well-characterized, the phagocytic and synaptic pruning roles of microglia in neuropathic pain remain unknown. Here, we show that peripheral nerve injury induces engulfment of dorsal horn synapses by microglia, leading to a preferential loss of inhibitory synapses. This synapse removal is dependent on the microglial complement-mediated synapse pruning pathway, as mice deficient in complement C3 do not exhibit synapse elimination. Furthermore, pharmacological inhibition of the complement protein C1q prevents dorsal horn inhibitory synapse loss and attenuates neuropathic pain. Thus, these results demonstrate that the complement pathway promotes persistent pain hypersensitivity via microglia-mediated engulfment and loss of inhibitory synapses in the dorsal horn of the spinal cord, revealing C1q as a novel therapeutic target in neuropathic pain.

[1]  Christopher W. Whelan,et al.  Overexpression of schizophrenia susceptibility factor human complement C4A promotes excessive synaptic loss and behavioral changes in mice , 2020, Nature Neuroscience.

[2]  R. Jahn,et al.  Local externalization of phosphatidylserine mediates developmental synaptic pruning by microglia , 2020, bioRxiv.

[3]  S. Prescott,et al.  Excitatory neurons are more disinhibited than inhibitory neurons by chloride dysregulation in the spinal dorsal horn , 2019, bioRxiv.

[4]  M. Greenberg,et al.  Sensory lesioning induces microglial synapse elimination via ADAM10 and fractalkine signaling , 2019, Nature Neuroscience.

[5]  Loren J. Martin,et al.  Genetic pathway analysis reveals a major role for extracellular matrix organization in inflammatory and neuropathic pain , 2019, Pain.

[6]  Y. Qadri,et al.  Microglia in Pain: Detrimental and Protective Roles in Pathogenesis and Resolution of Pain , 2018, Neuron.

[7]  Z. Borhegyi,et al.  Local apoptotic-like mechanisms underlie complement-mediated synaptic pruning , 2018, Proceedings of the National Academy of Sciences.

[8]  Y. Yanagawa,et al.  NMDA Receptor Activation Underlies the Loss of Spinal Dorsal Horn Neurons and the Transition to Persistent Pain after Peripheral Nerve Injury , 2018, Cell reports.

[9]  T. D. Kweon,et al.  Gabapentin prevents synaptogenesis between sensory and spinal cord neurons induced by thrombospondin‐4 acting on pre‐synaptic Cavα2δ1 subunits and involving T‐type Ca2+ channels , 2018, British journal of pharmacology.

[10]  M. Tsuda,et al.  Microglia in neuropathic pain: cellular and molecular mechanisms and therapeutic potential , 2018, Nature Reviews Neuroscience.

[11]  C. Gross,et al.  Errant gardeners: glial-cell-dependent synaptic pruning and neurodevelopmental disorders , 2017, Nature Reviews Neuroscience.

[12]  Y. de Koninck,et al.  Spinal microglia are required for long-term maintenance of neuropathic pain , 2017, Pain.

[13]  Herta Flor,et al.  Structural plasticity and reorganisation in chronic pain , 2017, Nature Reviews Neuroscience.

[14]  Rebecca P Seal,et al.  Neural circuits for pain: Recent advances and current views , 2016, Science.

[15]  Yong Ho Kim,et al.  Spinal inhibition of p38 MAP kinase reduces inflammatory and neuropathic pain in male but not female mice: Sex-dependent microglial signaling in the spinal cord , 2016, Brain, Behavior, and Immunity.

[16]  Hong Wang,et al.  TMEM16F Regulates Spinal Microglial Function in Neuropathic Pain States , 2016, Cell reports.

[17]  D. Selkoe,et al.  Complement and microglia mediate early synapse loss in Alzheimer mouse models , 2016, Science.

[18]  Michelle K. Cahill,et al.  Progranulin Deficiency Promotes Circuit-Specific Synaptic Pruning by Microglia via Complement Activation , 2016, Cell.

[19]  Yong Ho Kim,et al.  High-resolution transcriptome analysis reveals neuropathic pain gene-expression signatures in spinal microglia after nerve injury , 2016, Pain.

[20]  Grayson O. Sipe,et al.  Microglial P2Y12 is necessary for synaptic plasticity in mouse visual cortex , 2016, Nature Communications.

[21]  Giulio Genovese,et al.  Schizophrenia risk from complex variation of complement component 4 , 2016, Nature.

[22]  Loren J. Martin,et al.  Different immune cells mediate mechanical pain hypersensitivity in male and female mice , 2015, Nature Neuroscience.

[23]  M. Tsuda.  Microglia in the spinal cord and neuropathic pain , 2015, Journal of diabetes investigation.

[24]  Idrish Ali,et al.  Role of fractalkine–CX3CR1 pathway in seizure-induced microglial activation, neurodegeneration, and neuroblast production in the adult rat brain , 2015, Neurobiology of Disease.

[25]  J. A. Payne,et al.  Cation-chloride cotransporters in neuronal development, plasticity and disease , 2014, Nature Reviews Neuroscience.

[26]  Emily K. Lehrman,et al.  An engulfment assay: a protocol to assess interactions between CNS phagocytes and neurons. , 2014, Journal of visualized experiments : JoVE.

[27]  Björn Usadel,et al.  Trimmomatic: a flexible trimmer for Illumina sequence data , 2014, Bioinform..

[28]  M. Malcangio,et al.  Fractalkine/CX3CR1 signaling during neuropathic pain , 2014, Front. Cell. Neurosci..

[29]  V. Granados-Soto,et al.  Role of spinal P2Y6 and P2Y11 receptors in neuropathic pain in rats: possible involvement of glial cells , 2014, Molecular pain.

[30]  K. J. Murphy,et al.  Neuron–glia crosstalk in health and disease: fractalkine and CX3CR1 take centre stage , 2013, Open Biology.

[31]  Beth Stevens,et al.  TGF-β Signaling Regulates Neuronal C1q Expression and Developmental Synaptic Refinement , 2013, Nature Neuroscience.

[32]  E. Huang,et al.  A Dramatic Increase of C1q Protein in the CNS during Normal Aging , 2013, The Journal of Neuroscience.

[33]  Cole Trapnell,et al.  TopHat2: accurate alignment of transcriptomes in the presence of insertions, deletions and gene fusions , 2013, Genome Biology.

[34]  Reinhard Schneider,et al.  Nuclear Calcium Signaling in Spinal Neurons Drives a Genomic Program Required for Persistent Inflammatory Pain , 2013, Neuron.

[35]  David G Hendrickson,et al.  Differential analysis of gene regulation at transcript resolution with RNA-seq , 2012, Nature Biotechnology.

[36]  Ben A. Barres,et al.  Microglia Sculpt Postnatal Neural Circuits in an Activity and Complement-Dependent Manner , 2012, Neuron.

[37]  J. Mogil,et al.  Spinal Cord Toll-Like Receptor 4 Mediates Inflammatory and Neuropathic Hypersensitivity in Male But Not Female Mice , 2011, The Journal of Neuroscience.

[38]  M. Giustetto,et al.  Synaptic Pruning by Microglia Is Necessary for Normal Brain Development , 2011, Science.

[39]  H. Tozaki-Saitoh,et al.  Nerve injury‐activated microglia engulf myelinated axons in a P2Y12 signaling‐dependent manner in the dorsal horn , 2010, Glia.

[40]  G. Wasner,et al.  Neuropathic pain: diagnosis, pathophysiological mechanisms, and treatment , 2010, The Lancet Neurology.

[41]  Alban Latremoliere,et al.  Central sensitization: a generator of pain hypersensitivity by central neural plasticity. , 2009, The journal of pain : official journal of the American Pain Society.

[42]  Michael Costigan,et al.  Neuropathic pain: a maladaptive response of the nervous system to damage. , 2009, Annual review of neuroscience.

[43]  Cole Trapnell,et al.  Ultrafast and memory-efficient alignment of short DNA sequences to the human genome , 2009, Genome Biology.

[44]  M. Gustafsson,et al.  Three-dimensional resolution doubling in wide-field fluorescence microscopy by structured illumination. , 2008, Biophysical journal.

[45]  John D. Lambris,et al.  The Classical Complement Cascade Mediates CNS Synapse Elimination , 2007, Cell.

[46]  Clifford J. Woolf,et al.  Complement Induction in Spinal Cord Microglia Results in Anaphylatoxin C5a-Mediated Pain Hypersensitivity , 2007, The Journal of Neuroscience.

[47]  K. Jacobson,et al.  UDP acting at P2Y6 receptors is a mediator of microglial phagocytosis , 2007, Nature.

[48]  C. Gravel,et al.  BDNF from microglia causes the shift in neuronal anion gradient underlying neuropathic pain , 2005, Nature.

[49]  A. Basbaum,et al.  Spared nerve injury model of neuropathic pain in the mouse: a behavioral and anatomic analysis. , 2003, The journal of pain : official journal of the American Pain Society.

[50]  Yves De Koninck,et al.  Trans-synaptic shift in anion gradient in spinal lamina I neurons as a mechanism of neuropathic pain , 2003, Nature.

[51]  S. Alam,et al.  CX3CR1 Tyrosine Sulfation Enhances Fractalkine-induced Cell Adhesion* , 2002, The Journal of Biological Chemistry.

[52]  C. Woolf,et al.  Spared nerve injury: an animal model of persistent peripheral neuropathic pain , 2000, Pain.

[53]  R. Skinner,et al.  Intraspinal non‐neuronal cellular responses to peripheral nerve injury , 1979, The Anatomical record.

[54]  A. Bialas,et al.  Complement System in Neural Synapse Elimination in Development and Disease. , 2017, Advances in immunology.

[55]  D. Wilton,et al.  Structured Illumination Microscopy for the Investigation of Synaptic Structure and Function. , 2017, Methods in molecular biology.

[56]  B. Peterson,et al.  Normal Development of Brain Circuits , 2010, Neuropsychopharmacology.

[57]  J. Heavner.  The spinal cord dorsal horn. , 1973, Anesthesiology.