Using automated patch clamp electrophysiology platforms in pain‐related ion channel research: insights from industry and academia

Bell, Damian C.; Dallas, Mark L.

doi:10.1111/bph.13916

Cited by 47 publications

(32 citation statements)

References 77 publications

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“…This manuscript presents results from studies on four prominent human cardiac currents that modulate cardiac repolarization (hERG, peak hNav1. 5, and, to a lesser extent, hCav1.2 and late hNav1.5). Results are focused on two currents (hERG and peak hNav1.5) as the most data were collected on these currents.…”

mentioning

confidence: 89%

Cross-site and cross-platform variability of automated patch clamp assessments of drug effects on human cardiac currents in recombinant cells

Kramer

Himmel

Lindqvist³

et al. 2020

Sci Rep

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Automated patch clamp (APC) instruments enable efficient evaluation of electrophysiologic effects of drugs on human cardiac currents in heterologous expression systems. Differences in experimental protocols, instruments, and dissimilar site procedures affect the variability of IC 50 values characterizing drug block potency. this impacts the utility of Apc platforms for assessing a drug's cardiac safety margin. We determined variability of Apc data from multiple sites that measured blocking potency of 12 blinded drugs (with different levels of proarrhythmic risk) against four human cardiac currents (heRG [i Kr ], hCav1.2 [L-Type I ca ], peak hNav1.5, [Peak I na ], late hNav1.5 [Late I na ]) with recommended protocols (to minimize variance) using five APC platforms across 17 sites. IC 50 variability (25/75 percentiles) differed for drugs and currents (e.g., 10.4-fold for dofetilide block of hERG current and 4-fold for mexiletine block of hNav1.5 current). Within-platform variance predominated for 4 of 12 hERG blocking drugs and 4 of 6 hNav1.5 blocking drugs. hERG and hNav1.5 block. Bland-Altman plots depicted varying agreement across Apc platforms. A follow-up survey suggested multiple sources of experimental variability that could be further minimized by stricter adherence to standard protocols. Adoption of best practices would ensure less variable Apc datasets and improved safety margins and proarrhythmic risk assessments. The characterization of drug effects on human currents using automated patch clamp (APC) platforms employing single-or multi-hole planar patch techniques 1-5 has revolutionized the assessment of electrophysiologic (and potential proarrhythmic) effects of new drug candidates. This increasingly popular experimental approach

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mentioning

confidence: 89%

Cross-site and cross-platform variability of automated patch clamp assessments of drug effects on human cardiac currents in recombinant cells

Kramer

Himmel

Lindqvist³

et al. 2020

Sci Rep

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“…The experiments were carried out at constant room temperature (22°C).2.8. Automated patch-clamp recordings from HEK-293 and CHO cellsWhole-cell automated patch-clamp experiments were performed on a QPatch HTX automated electrophysiology platform (Sophion Bioscience, Ballerup, Denmark), allowing both signal acquisition and data analyses(Bell and Dallas, 2017). The extracellular medium composition of hNa V -overexpressing HEK293 cells was (in mM): NaCl 154, KCl 4, CaCl 2 2, MgCl 2 1, and HEPES 10 (pH 7.4, adjusted with NaOH), and that of intracellular (i.e.…”

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confidence: 99%

Direct evidence for high affinity blockade of NaV1.6 channel subtype by huwentoxin-IV spider peptide, using multiscale functional approaches

et al. 2018

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The Chinese bird spider huwentoxin-IV (HwTx-IV) is well-known to be a highly potent blocker of Na1.7 subtype of voltage-gated sodium (Na) channels, a genetically validated analgesic target, and thus promising as a potential lead molecule for the development of novel pain therapeutics. In the present study, the interaction between HwTx-IV and Na1.6 channel subtype was investigated using multiscale (from in vivo to individual cell) functional approaches. HwTx-IV was approximatively 2 times more efficient than tetrodotoxin (TTX) to inhibit the compound muscle action potential recorded from the mouse skeletal neuromuscular system in vivo, and 30 times more effective to inhibit nerve-evoked than directly-elicited muscle contractile force of isolated mouse hemidiaphragms. These results strongly suggest that the inhibition of nerve-evoked skeletal muscle functioning, produced by HwTx-IV, resulted from a toxin-induced preferential blockade of Na1.6, compared to Na1.4, channel subtype. This was confirmed by whole-cell automated patch-clamp experiments performed on human embryonic kidney (HEK)-293 cells overexpressing hNa1.1-1.8 channel subtypes. HwTx-IV was also approximatively 850 times more efficient to inhibit TTX-sensitive than TTX-resistant sodium currents recorded from mouse dorsal root ganglia neurons. Finally, based on our data, we predict that blockade of the Na1.6 channel subtype was involved in the in vivo toxicity of HwTx-IV, although this toxicity was more than 2 times lower than that of TTX. In conclusion, our results provide detailed information regarding the effects of HwTx-IV and allow a better understanding of the side-effect mechanisms involved in vivo and of channel subtype interactions resulting from the toxin activity.

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“…Such model systems, theoretically, allow ion channel pharmacology and/or biophysics to be examined in isolation. These efforts are supported by developments in automated patch clamp technology, predominantly using expression systems, to measure ion channel activity (Bell and Dallas, ; Loucif et al ., ).…”

Section: Measuring Ion Channel Function In Nociceptive Pathwaysmentioning

confidence: 99%

Recent advances in targeting ion channels to treat chronic pain

Stevens¹,

Stephens

2018

British J Pharmacology

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Using automated patch clamp electrophysiology platforms in pain‐related ion channel research: insights from industry and academia

Abstract: This article is part of a themed section on Recent Advances in Targeting Ion Channels to Treat Chronic Pain. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v175.12/issuetoc.

Cited by 47 publications

References 77 publications

Cross-site and cross-platform variability of automated patch clamp assessments of drug effects on human cardiac currents in recombinant cells

Cross-site and cross-platform variability of automated patch clamp assessments of drug effects on human cardiac currents in recombinant cells

Direct evidence for high affinity blockade of NaV1.6 channel subtype by huwentoxin-IV spider peptide, using multiscale functional approaches

Recent advances in targeting ion channels to treat chronic pain

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