RgIA4 Prevention of Acute Oxaliplatin-Induced Cold Allodynia Requires α9-Containing Nicotinic Acetylcholine Receptors and CD3+ T-Cells

Chemotherapy-induced neuropathic pain is a debilitating and dose-limiting side effect. Oxaliplatin is a third-generation platinum and antineoplastic compound that is commonly used to treat colorectal cancer and commonly yields neuropathic side effects. Available drugs such as duloxetine provide only modest benefits against oxaliplatin-induced neuropathy. A particularly disruptive symptom of oxaliplatin is painful cold sensitivity, known as cold allodynia. Previous studies of the Conus regius peptide, RgIA, and its analogs have demonstrated relief from oxaliplatin-induced cold allodynia, yielding improvement that persists even after treatment cessation. Moreover, underlying inflammatory and neuronal protection were shown at the cellular level in chronic constriction nerve injury models, consistent with disease-modifying effects. Despite these promising preclinical outcomes, the underlying molecular mechanism of action of RgIA4 remains an area of active investigation. This study aimed to determine the necessity of the α9 nAChR subunit and potential T-cell mechanisms in RgIA4 efficacy against acute oxaliplatin-induced cold allodynia. A single dose of oxaliplatin (10 mg/kg) was utilized followed by four daily doses of RgIA4. Subcutaneous administration of RgIA4 (40 µg/kg) prevented cold allodynia in wildtype mice but not in mice lacking the α9 nAChR-encoding gene, chrna9. RgIA4 also failed to reverse allodynia in mice depleted of CD3+ T-cells. In wildtype mice treated with oxaliplatin, quantitated circulating T-cells remained unaffected by RgIA4. Together, these results show that RgIA4 requires both chrna9 and CD3+ T-cells to exert its protective effects against acute cold-allodynia produced by oxaliplatin.

[1]  J. McIntosh,et al.  Alkaloid ligands enable function of homomeric human α10 nicotinic acetylcholine receptors , 2022, Frontiers in Pharmacology.

[2]  O. Kaminuma,et al.  Expression and Function of Nicotinic Acetylcholine Receptors in Induced Regulatory T Cells , 2022, International journal of molecular sciences.

[3]  A. Nicke,et al.  Interaction of α9α10 Nicotinic Receptors With Peptides and Proteins From Animal Venoms , 2021, Frontiers in Cellular Neuroscience.

[4]  David John Adams,et al.  Analgesic α‐conotoxins modulate native and recombinant GIRK1/2 channels via activation of GABAB receptors and reduce neuroexcitability , 2021, British journal of pharmacology.

[5]  J. McIntosh,et al.  Discovery of Methylene Thioacetal-Incorporated α-RgIA Analogues as Potent and Stable Antagonists of the Human α9α10 Nicotinic Acetylcholine Receptor for the Treatment of Neuropathic Pain. , 2021, Journal of medicinal chemistry.

[6]  J. McIntosh,et al.  Selective Penicillamine Substitution Enables Development of a Potent Analgesic Peptide that Acts through a Non-Opioid-Based Mechanism. , 2021, Journal of medicinal chemistry.

[7]  R. Norton,et al.  Alkyne-Bridged α-Conotoxin Vc1.1 Potently Reverses Mechanical Allodynia in Neuropathic Pain Models. , 2021, Journal of Medicinal Chemistry.

[8]  A. Kavelaars,et al.  T Cells as Guardians of Pain Resolution. , 2021, Trends in molecular medicine.

[9]  J. McIntosh,et al.  Development of Conformationally Constrained α-RgIA Analogues as Stable Peptide Antagonists of Human α9α10 Nicotinic Acetylcholine Receptors. , 2020, Journal of medicinal chemistry.

[10]  D. Craik,et al.  Critical Residue Properties for Potency and Selectivity of α-Conotoxin RgIA Towards α9α10 Nicotinic Acetylcholine Receptors. , 2020, Biochemical pharmacology.

[11]  Stephanie I. Shiers,et al.  Quantitative differences in neuronal subpopulations between mouse and human dorsal root ganglia demonstrated with RNAscope in situ hybridization. , 2020, Pain.

[12]  R. Su,et al.  The α9α10 Nicotinic Acetylcholine Receptor Antagonist αO-Conotoxin GeXIVA[1,2] Alleviates and Reverses Chemotherapy-Induced Neuropathic Pain , 2019, Marine drugs.

[13]  V. Apostolopoulos,et al.  Oxaliplatin Treatment Alters Systemic Immune Responses , 2019, BioMed research international.

[14]  J. McIntosh,et al.  Conopeptides [V11L;V16D]ArIB and RgIA4: Powerful Tools for the Identification of Novel Nicotinic Acetylcholine Receptors in Monocytes , 2019, Front. Pharmacol..

[15]  P. Dougherty,et al.  Beyond symptomatic relief for chemotherapy‐induced peripheral neuropathy: Targeting the source , 2018, Cancer.

[16]  D. Servent,et al.  α9‐containing nicotinic acetylcholine receptors and the modulation of pain , 2018, British journal of pharmacology.

[17]  D. Craik,et al.  Structure-Activity Studies Reveal the Molecular Basis for GABAB-Receptor Mediated Inhibition of High Voltage-Activated Calcium Channels by α-Conotoxin Vc1.1. , 2018, ACS chemical biology.

[18]  B. Morley,et al.  Commentary: Nicotinic Acetylcholine Receptor α9 and α10 Subunits Are Expressed in the Brain of Mice , 2018, Front. Cell. Neurosci..

[19]  A. Scope,et al.  NKp46 Receptor‐Mediated Interferon‐&ggr; Production by Natural Killer Cells Increases Fibronectin 1 to Alter Tumor Architecture and Control Metastasis , 2018, Immunity.

[20]  L. Basso,et al.  T-lymphocyte-derived enkephalins reduce Th1/Th17 colitis and associated pain in mice , 2018, Journal of Gastroenterology.

[21]  M. Zucchetti,et al.  Susceptibility of different mouse strains to oxaliplatin peripheral neurotoxicity: Phenotypic and genotypic insights , 2017, PloS one.

[22]  E. Glowatzki,et al.  RgIA4 Potently Blocks Mouse α9α10 nAChRs and Provides Long Lasting Protection against Oxaliplatin-Induced Cold Allodynia , 2017, Front. Cell. Neurosci..

[23]  W. Padberg,et al.  Canonical and Novel Non-Canonical Cholinergic Agonists Inhibit ATP-Induced Release of Monocytic Interleukin-1β via Different Combinations of Nicotinic Acetylcholine Receptor Subunits α7, α9 and α10 , 2017, Front. Cell. Neurosci..

[24]  C. Ghelardini,et al.  Inhibition of α9α10 nicotinic acetylcholine receptors prevents chemotherapy-induced neuropathic pain , 2017, Proceedings of the National Academy of Sciences.

[25]  D. Goldstein,et al.  Characterisation of Immune and Neuroinflammatory Changes Associated with Chemotherapy-Induced Peripheral Neuropathy , 2017, PloS one.

[26]  C. Ghelardini,et al.  The α9α10 nicotinic receptor antagonist α-conotoxin RgIA prevents neuropathic pain induced by oxaliplatin treatment , 2016, Experimental Neurology.

[27]  W. Padberg,et al.  Phosphocholine – an agonist of metabotropic but not of ionotropic functions of α9-containing nicotinic acetylcholine receptors , 2016, Scientific Reports.

[28]  D. Craik,et al.  Cloning, synthesis, and characterization of αO-conotoxin GeXIVA, a potent α9α10 nicotinic acetylcholine receptor antagonist , 2015, Proceedings of the National Academy of Sciences.

[29]  K. Vowles,et al.  Rates of opioid misuse, abuse, and addiction in chronic pain: a systematic review and data synthesis , 2015, Pain.

[30]  David John Adams,et al.  Novel Mechanism of Voltage-Gated N-type (Cav2.2) Calcium Channel Inhibition Revealed through α-Conotoxin Vc1.1 Activation of the GABAB Receptor , 2015, Molecular Pharmacology.

[31]  A. Cesario,et al.  Alpha9Alpha10 Nicotinic Acetylcholine Receptors as Target for the Treatment of Chronic Pain , 2014 .

[32]  M. J. McIntosh,et al.  α-Conotoxin RgIA protects against the development of nerve injury-induced chronic pain and prevents both neuronal and glial derangement , 2014, PAIN®.

[33]  Gila Moalem-Taylor,et al.  Regulatory T cells attenuate neuropathic pain following peripheral nerve injury and experimental autoimmune neuritis , 2012, PAIN®.

[34]  D. Bennett,et al.  The role of the immune system in the generation of neuropathic pain , 2012, The Lancet Neurology.

[35]  A. Eschalier,et al.  Oxaliplatin-induced cold hypersensitivity is due to remodelling of ion channel expression in nociceptors , 2011, EMBO molecular medicine.

[36]  A. Viola,et al.  Differential involvement of α4β2, α7 and α9α10 nicotinic acetylcholine receptors in B lymphocyte activation in vitro. , 2011, The international journal of biochemistry & cell biology.

[37]  K. Huth,et al.  CD4(+) T-cells are important in regulating macrophage polarization in C57BL/6 wild-type mice. , 2011, Cellular immunology.

[38]  Maria Fitzgerald,et al.  T-Cell Infiltration and Signaling in the Adult Dorsal Spinal Cord Is a Major Contributor to Neuropathic Pain-Like Hypersensitivity , 2009, The Journal of Neuroscience.

[39]  M. Friedlander,et al.  Oxaliplatin-induced neurotoxicity: changes in axonal excitability precede development of neuropathy. , 2009, Brain : a journal of neurology.

[40]  R. Baron,et al.  Oxaliplatin-induced painful neuropathy – Flicker of hope or hopeless pain? , 2009, Pain.

[41]  C. Loprinzi,et al.  Chemotherapy-induced peripheral neuropathy: prevention and treatment strategies. , 2008, European journal of cancer.

[42]  R. Norton,et al.  Alpha-RgIA, a novel conotoxin that blocks the alpha9alpha10 nAChR: structure and identification of key receptor-binding residues. , 2008, Journal of molecular biology.

[43]  P. Kisielow,et al.  Identification of Genes Involved in Positive Selection of CD4+8+ Thymocytes: Expanding the Inventory , 2007, Immunological investigations.

[44]  J. McIntosh,et al.  Molecular mechanism for analgesia involving specific antagonism of α9α10 nicotinic acetylcholine receptors , 2006, Proceedings of the National Academy of Sciences.

[45]  J. McIntosh,et al.  α-RgIA: A Novel Conotoxin That Specifically and Potently Blocks the α9α10 nAChR†,‡ , 2006 .

[46]  K. Gayler,et al.  Alpha-conotoxin Vc1.1 alleviates neuropathic pain and accelerates functional recovery of injured neurones , 2005, Brain Research.

[47]  R. J. Cersosimo Oxaliplatin-Associated Neuropathy: A Review , 2005, The Annals of pharmacotherapy.

[48]  G. Moalem,et al.  T lymphocytes play a role in neuropathic pain following peripheral nerve injury in rats , 2004, Neuroscience.

[49]  R. Ferris,et al.  Characterization of the human nicotinic acetylcholine receptor subunit alpha (alpha) 9 (CHRNA9) and alpha (alpha) 10 (CHRNA10) in lymphocytes. , 2004, Life sciences.

[50]  S. Heinemann,et al.  α10: A determinant of nicotinic cholinergic receptor function in mammalian vestibular and cochlear mechanosensory hair cells , 2001, Proceedings of the National Academy of Sciences of the United States of America.

[51]  Jacques Barhanin,et al.  Role of α9 Nicotinic ACh Receptor Subunits in the Development and Function of Cochlear Efferent Innervation , 1999, Neuron.

[52]  S. Heinemann,et al.  α9: An acetylcholine receptor with novel pharmacological properties expressed in rat cochlear hair cells , 1994, Cell.