Suppression of inflammatory and neuropathic pain by uncoupling CRMP-2 from the presynaptic Ca2+ channel complex

The use of N-type voltage-gated calcium channel (CaV2.2) blockers to treat pain is limited by many physiological side effects. Here we report that inflammatory and neuropathic

hypersensitivity can be suppressed by inhibiting the binding of collapsin response mediator protein 2 (CRMP-2) to CaV2.2 and thereby reducing channel function. A peptide of CRMP-2 fused to

the HIV transactivator of transcription (TAT) protein (TAT-CBD3) decreased neuropeptide release from sensory neurons and excitatory synaptic transmission in dorsal horn neurons, reduced

meningeal blood flow, reduced nocifensive behavior induced by formalin injection or corneal capsaicin application and reversed neuropathic hypersensitivity produced by an antiretroviral

drug. TAT-CBD3 was mildly anxiolytic without affecting memory retrieval, sensorimotor function or depression. At doses tenfold higher than that required to reduce hypersensitivity in vivo,

TAT-CBD3 caused a transient episode of tail kinking and body contortion. By preventing CRMP-2–mediated enhancement of CaV2.2 function, TAT-CBD3 alleviated inflammatory and neuropathic

hypersensitivity, an approach that may prove useful in managing chronic pain.

This work is supported by grants from the US National Institutes of Health: Dental and Craniofacial Research (DE14318-06 to J.C.F. and DE017794 to R.-R.J.), Drug Abuse (DA026040 to F.A.W.),

Neurological Disorders and Stroke (NS051668 to C.M.H. and NS050131 to N.B.) and Environmental Health Sciences (ES017430 to G.S.O. and J.H.H.); the Indiana State Department of Health−Spinal

Cord and Brain Injury Fund (A70-0-079212 to N.B. and A70-9-079138 to R.K.) and the Indiana University Biomedical Committee–Research Support Funds (2286501 to R.K.); a National Scientist

Development Grant from the American Heart Association (SDG5280023 to R.K.); and the Elwert Award in Medicine to R.K. J.M.B. is the recipient of a Larry Kays Medical Neuroscience fellowship.

S.M.W. is a Stark Scholar. We thank A. Molosh and members of the Pain and Sensory Group for discussions, S.K. Ahuja for assistance with behavioral experiments and C. Kohn for comments on the

manuscript.

Joel M Brittain and Djane B Duarte: These authors contributed equally to this work.

Program in Medical Neurosciences, Paul and Carole Stark Neurosciences Research Institute, Indiana University School of Medicine, Indianapolis, Indiana, USA

Joel M Brittain, Djane B Duarte, Sarah M Wilson, Weiguo Zhu, Carrie Ballard, Naikui Liu, Wenhui Xiong, Matthew S Ripsch, Yuying Wang, Jill C Fehrenbacher, Brian S Schmutzler, Bo Myung Cheon,

 Michael R Due, Nicole M Ashpole, Andy Hudmon, Samy O Meroueh, Cynthia M Hingtgen, Nickolay Brustovetsky, Joyce H Hurley, Xiaoming Jin, Xiao-Ming Xu, Gerry S Oxford, Michael R Vasko, 

Fletcher A White & Rajesh Khanna

Department of Pharmacology and Toxicology, Indiana University School of Medicine, Indianapolis, Indiana, USA

Weiguo Zhu, Yuying Wang, Jill C Fehrenbacher, Brian S Schmutzler, Tatiana Brustovetsky, Andy Hudmon, Cynthia M Hingtgen, Nickolay Brustovetsky, Gerry S Oxford, Michael R Vasko & Rajesh

Khanna

Department of Biochemistry and Molecular Biology, Indiana University School of Medicine, Indianapolis, Indiana, USA

Carrie Ballard, May Khanna, Nicole M Ashpole, Andy Hudmon, Samy O Meroueh & Joyce H Hurley

Department of Psychiatry, Indiana University School of Medicine, Indianapolis, Indiana, USA

Department of Neurological Surgery, Indiana University School of Medicine, Indianapolis, Indiana, USA

Department of Anatomy and Cell Biology, Indiana University School of Medicine, Indianapolis, Indiana, USA

Department of Anesthesia, Indiana University School of Medicine, Indianapolis, Indiana, USA

Matthew S Ripsch, Jill C Fehrenbacher, Bo Myung Cheon, Michael R Due, Michael R Vasko & Fletcher A White

Department of Anesthesiology, Sensory Plasticity Laboratory, Pain Research Center, Perioperative and Pain Medicine, Brigham and Women's Hospital and Harvard Medical School, Boston,

Massachusetts, USA

Department of Neurology, Indiana University School of Medicine, Indianapolis, Indiana, USA

Department of Indiana Clinical and Translational Sciences Institute, Indiana University School of Medicine, Indianapolis, Indiana, USA

J.M.B. performed molecular biology, biochemistry and calcium imaging experiments and analyzed the data. D.B.D. carried out the spinal cord slice release and formalin behavior experiments and

helped to write the manuscript. S.M.W. performed immunocytochemistry and wrote the manuscript. C.B. carried out the laser Doppler blood flowmetry. J.H.H. analyzed the blood flow data.

P.L.J. and S.D.F. performed anxiety and despair behavior experiments. W.Z. and Y.W. performed DRG and hippocampal patching. C.-K.P. conducted electrophysiology in spinal cord slices. W.X.

and X.J. performed electrophysiology on brain slices. B.S.S. carried out the DRG release assays. T.B., N.B. and J.M.B. performed and analyzed the calcium imaging experiments. B.M.C., M.R.D.

and M.S.R. performed DRG immunocytochemistry and ddC behavior experiments. M.K. and S.O.M. performed the surface plasmon resonance experiments and analyzed the data. N.L. performed the

rotarod and water maze experiments. J.C.F. performed the nocifensive behavior experiments and editing of the manuscript. N.M.A. and A.H. synthesized the peptide blot. X.-M.X., C.M.H.,

M.R.V., G.S.O. and A.S. contributed to editing of the manuscript. R.-R.J contributed to electrophysiology of spinal cord slices and editing of the manuscript. F.A.W. analyzed the ddC

behavior data and contributed to writing and editing the manuscript. R.K. identified the peptide, conceived the study, designed and supervised the overall project and wrote the manuscript.

Supplementary Figures 1–7, Supplementary Table 1 and Supplementary Methods (PDF 2807 kb)

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