Potent Modulation of the Voltage-Gated Sodium Channel Na<sub>v</sub>1.7 by OD1, a Toxin from the Scorpion <i>Odonthobuthus doriae</i>

Maertens, Chantal; Cuypers, Eva; Amininasab, Mehriar; Jalali, Amir; Vatanpour, Hossein; Tytgat, Jan

doi:10.1124/mol.106.022970

Cited by 83 publications

(91 citation statements)

References 43 publications

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“…This demonstrates that the delay in Nav1.7 inactivation induced by OD1, in combination with a shift in the voltage-dependence of Nav activation elicited by veratridine, generates Ca 2+ influx that is mediated solely through Nav1.7. These results confirm the preferential activation of endogenous Nav1.7 over Nav1.2 by OD1 [10,23] and permit assessment of human Nav1.7 responses in an endogenous context without the need for addition of antagonists. At high concentrations, OD1 also effects a hyperpolarizing shift in the voltage of activation of Nav1.7 [23], which presumably forms the basis for the small Ca 2+ transients elicited by OD1 at high (> 30 nM) but not lower concentrations (data not shown).…”

supporting

confidence: 72%

“…These results confirm the preferential activation of endogenous Nav1.7 over Nav1.2 by OD1 [10,23] and permit assessment of human Nav1.7 responses in an endogenous context without the need for addition of antagonists. At high concentrations, OD1 also effects a hyperpolarizing shift in the voltage of activation of Nav1.7 [23], which presumably forms the basis for the small Ca 2+ transients elicited by OD1 at high (> 30 nM) but not lower concentrations (data not shown). In contrast, at low concentrations OD1 affects the gating of Nav1.7 and specifically impairs fast inactivation [10,23], so that small shifts in the voltagedependence of activation induced by veratridine result in synergistic activation of Nav1.7.…”

supporting

confidence: 72%

“…also effects a hyperpolarizing shift in the voltage of activation of Nav1.7 [23], which presumably forms the basis for the small Ca 2+ transients elicited by OD1 at high (> 30 nM) but not lower concentrations (data not shown). In contrast, at low concentrations OD1 affects the gating of Nav1.7 and specifically impairs fast inactivation [10,23], so that small shifts in the voltagedependence of activation induced by veratridine result in synergistic activation of Nav1.7.…”

Section: Discussionmentioning

confidence: 91%

“…As an alternative approach to generating Nav1.7 specific responses, we investigated the potential of the scorpion toxin OD1, which affects inactivation of Nav1.7 with high specificity [23], to synergise with veratridine. Treatment with OD1 caused a marked shift in the veratridine concentration-response curve producing responses that are now fully blocked by ProTxII.…”

Section: Discussionmentioning

confidence: 99%

“…Thus, this assay is amenable to the measurement of a diverse range of modulators of Nav function via detection of changes in Ca 2+ signalling. Using the industry-19 standard FLIPR platform in 96-or 384-well format we achieved a high signal-to-noise ratio (Z' score of 0.7), making this assay suitable for rapid identification of novel Nav blockers.As an alternative approach to generating Nav1.7 specific responses, we investigated the potential of the scorpion toxin OD1, which affects inactivation of Nav1.7 with high specificity [23], to synergise with veratridine. Treatment with OD1 caused a marked shift in the veratridine concentration-response curve producing responses that are now fully blocked by ProTxII.…”

mentioning

confidence: 99%

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Characterisation of Nav types endogenously expressed in human SH-SY5Y neuroblastoma cells

Vetter

Mozar

Durek

et al. 2012

Biochemical Pharmacology

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Abbreviations: Nav, voltage-gated sodium channel; TTX, tetrodotoxin; ProTxII, β/ω-theraphotoxin-Tp2a; Ca 2+ , calcium ion; DMEM, Dulbecco's Modified Eagle's Medium; FBS, foetal bovine serum; PBS, phosphate buffered saline; PSS, physiological salt solution; HBS, HEPES-buffered saline; AFU, arbitrary fluorescence unit; SEM, standard error of the mean; Fluo-4 AM, Fluo-4 acetoxymethylester; RPMI, Roswell Park Memorial Institute; BSA, bovine serum albumin; CCD, charge-coupled device; DAPI, 4',6-diamidino-2-phenylindole 2 AbstractThe human neuroblastoma cell line SH-SY5Y is a potentially useful model for the identification and characterisation of Nav modulators, but little is known about the pharmacology of their endogenously expressed Navs. The aim of this study was to determine the expression of endogenous Nav α and β subunits in SH-SY5Y cells using PCR and immunohistochemical approaches, and pharmacologically characterise the Nav isoforms endogenously expressed in this cell line using electrophysiological and fluorescence approaches. SH-SY5Y human neuroblastoma cells were found to endogenously express several Nav isoforms including Nav1.2 and Nav1.7. Activation of endogenously expressed Navs with veratridine or the scorpion toxin OD1 caused membrane depolarization and subsequent Ca 2+ influx through voltage-gated L-and N-type calcium channels, allowing Nav activation to be detected with membrane potential and fluorescent Ca 2 dyes. -Conotoxin TIIIA and ProTxII identified Nav1.2 and Nav1.7 as the major contributors of this response. The Nav1.7-selective scorpion toxin OD1 in combination with veratridine produced a Nav1.7-selective response, confirming that endogenously expressed human Nav1.7 in SH-SY5Y cells is functional and can be synergistically activated, providing a new assay format for ligand screening.Key Words: SH-SY5Y; Ca 2+ ; Nav1.7; ProTxII; OD1 3 IntroductionVoltage-gated sodium channels (Nav) are complex transmembrane proteins comprised of a poreforming α subunit and accessory β subunits that play an essential role in the initiation and propagation of action potentials in excitable cells. To date, apart from the related Nax which appears to function as a sodium sensor [1,2], nine isoforms termed Nav1.1 -Nav1.9 have been functionally defined as sodium-selective ion channels [3]. Their distinct tissue distribution and amenability to modulation by toxins and drugs has led to significant interest in Nav as therapeutic targets in a number of poorly treated conditions ranging from epilepsy to cardiac arrhythmias and pain [4]. In recent years Nav1.7 has emerged as an attractive drug target, with expression restricted to a subset of nociceptive neurons that is expected to limit on-target side effects of pharmacological modulators of Nav1.7 [5]. In addition, loss-of-function mutations in humans have highlighted the possibility that Nav1.7 inhibition could produce complete loss of pain sensations without dose-limiting side effects [6]. However, it remains unclear if such an effect can be translated to the clinic...

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supporting

confidence: 72%

supporting

confidence: 72%

Section: Discussionmentioning

confidence: 91%

Section: Discussionmentioning

confidence: 99%

mentioning

confidence: 99%

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Characterisation of Nav types endogenously expressed in human SH-SY5Y neuroblastoma cells

Vetter

Mozar

Durek

et al. 2012

Biochemical Pharmacology

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A Novel Antigen Design Strategy to Isolate Single‐Domain Antibodies that Target Human Nav1.7 and Reduce Pain in Animal Models

Martina,

Banderali,

Yogi

et al. 2024

Advanced Science

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Genetic studies have identified the voltage‐gated sodium channel 1.7 (Nav1.7) as pain target. Due to the ineffectiveness of small molecules and monoclonal antibodies as therapeutics for pain, single‐domain antibodies (VHHs) are developed against the human Nav1.7 (hNav1.7) using a novel antigen presentation strategy. A 70 amino‐acid peptide from the hNav1.7 protein is identified as a target antigen. A recombinant version of this peptide is grafted into the complementarity determining region 3 (CDR3) loop of an inert VHH in order to maintain the native 3D conformation of the peptide. This antigen is used to isolate one VHH able to i) bind hNav1.7, ii) slow the deactivation of hNav1.7, iii) reduce the ability of eliciting action potentials in nociceptors, and iv) reverse hyperalgesia in in vivo rat and mouse models. This VHH exhibits the potential to be developed as a therapeutic capable of suppressing pain. This novel antigen presentation strategy can be applied to develop biologics against other difficult targets such as ion channels, transporters and GPCRs.

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Advances in Targeting Voltage‐Gated Sodium Channels with Small Molecules

Nardi¹,

Damann²,

Hertrampf³

et al. 2012

ChemMedChem

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Blockade of voltage-gated sodium channels (VGSCs) has been used successfully in the clinic to enable control of pathological firing patterns that occur in conditions as diverse as chronic pain, epilepsy, and arrhythmias. Herein we review the state of the art in marketed sodium channel inhibitors, including a brief compendium of their binding sites and of the cellular and molecular biology of sodium channels. Despite the preferential action of this drug class toward over-excited cells, which significantly limits potential undesired side effects on other cells, the need to develop a second generation of sodium channel inhibitors to overcome their critical clinical shortcomings is apparent. Current approaches in drug discovery to deliver novel and truly innovative sodium channel inhibitors is next presented by surveying the most recent medicinal chemistry breakthroughs in the field of small molecules and developments in automated patch-clamp platforms. Various strategies aimed at identifying small molecules that target either particular isoforms of sodium channels involved in specific diseases or anomalous sodium channel currents, irrespective of the isoform by which they have been generated, are critically discussed and revised.

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Potent Modulation of the Voltage-Gated Sodium Channel Na_v1.7 by OD1, a Toxin from the Scorpion Odonthobuthus doriae

Abstract: Voltage-gated sodium channels are essential for the propagation of action potentials in nociceptive neurons.

Cited by 83 publications

References 43 publications

Characterisation of Nav types endogenously expressed in human SH-SY5Y neuroblastoma cells

Characterisation of Nav types endogenously expressed in human SH-SY5Y neuroblastoma cells

A Novel Antigen Design Strategy to Isolate Single‐Domain Antibodies that Target Human Nav1.7 and Reduce Pain in Animal Models

Advances in Targeting Voltage‐Gated Sodium Channels with Small Molecules

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Potent Modulation of the Voltage-Gated Sodium Channel Nav1.7 by OD1, a Toxin from the Scorpion Odonthobuthus doriae

Abstract: Voltage-gated sodium channels are essential for the propagation of action potentials in nociceptive neurons.

Cited by 83 publications

References 43 publications

Characterisation of Nav types endogenously expressed in human SH-SY5Y neuroblastoma cells

Characterisation of Nav types endogenously expressed in human SH-SY5Y neuroblastoma cells

A Novel Antigen Design Strategy to Isolate Single‐Domain Antibodies that Target Human Nav1.7 and Reduce Pain in Animal Models

Advances in Targeting Voltage‐Gated Sodium Channels with Small Molecules

Contact Info

Product

Resources

About

Potent Modulation of the Voltage-Gated Sodium Channel Na_v1.7 by OD1, a Toxin from the Scorpion Odonthobuthus doriae