Potent Inactivation-Dependent Inhibition of Adult and Neonatal NaV1.5 Channels by Lidocaine and Levobupivacaine

Taha Elajnaf (Lead / Corresponding author), Daniel T. Baptista-Hon, Tim G. Hales (Lead / Corresponding author)

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Abstract

Background: Cardiotoxic effects of local anesthetics (LAs) involve inhibition of Na V1.5 voltagegated Na + channels. Metastatic breast and colon cancer cells also express Na V1.5, predominantly the neonatal splice variant (nNa V1.5) and their inhibition by LAs reduces invasion and migration. It may be advantageous to target cancer cells while sparing cardiac function through selective blockade of nNa V1.5 and/or by preferentially affecting inactivated Na V1.5, which predominate in cancer cells. We tested the hypotheses that lidocaine and levobupivacaine differentially affect (1) adult (aNa V1.5) and nNa V1.5 and (2) the resting and inactivated states of Na V1.5. Methods: The whole-cell voltage-clamp technique was used to evaluate the actions of lidocaine and levobupivacaine on recombinant Na V1.5 channels expressed in HEK-293 cells. Cells were transiently transfected with cDNAs encoding either aNa V1.5 or nNa V1.5. Voltage protocols were applied to determine depolarizing potentials that either activated or inactivated 50% of maximum conductance (V1/2 activation and V1/2 inactivation, respectively). Results: Lidocaine and levobupivacaine potently inhibited aNa V1.5 (IC 50 mean [SD]: 20 [22] and 1 [0.6] μM, respectively) and nNa V1.5 (IC 50 mean [SD]: 17 [10] and 3 [1.6] μM, respectively) at a holding potential of -80 mV. IC 50s differed significantly between lidocaine and levobupivacaine with no influence of splice variant. Levobupivacaine induced a statistically significant depolarizing shift in the V1/2 activation for aNa V1.5 (mean [SD] from -32 [4.6] mV to -26 [8.1] mV) but had no effect on the voltage dependence of activation of nNa V1.5. Lidocaine had no effect on V1/2 activation of either variant but caused a significantly greater depression of maximum current mediated by nNa V1.5 compared to aNa V1.5. Similar statistically significant shifts in the V1/2 inactivation (approximately -10 mV) occurred for both LAs and Na V1.5 variants. Levobupivacaine (1 μM) caused a significantly greater slowing of recovery from inactivation of both variants than did lidocaine (10 μM). Both LAs caused approximately 50% tonic inhibition of aNa V1.5 or nNa V1.5 when holding at -80 mV. Neither LA caused tonic block at a holding potential of either -90 or -120 mV, voltages at which there was little steady-state inactivation. Higher concentrations of either lidocaine (300 μM) or levobupivacaine (100 μM) caused significantly more tonic block at -120 mV. Conclusions: These data demonstrate that low concentrations of the LAs exhibit inactivation-dependent block of Na V1.5, which may provide a rationale for their use to safely inhibit migration and invasion by metastatic cancer cells without cardiotoxicity.

Original languageEnglish
Pages (from-to)650-660
Number of pages11
JournalAnesthesia and Analgesia
Volume127
Issue number3
Early online date28 Jun 2018
DOIs
Publication statusPublished - Sep 2018

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Lidocaine
Local Anesthetics
Neoplasms
HEK293 Cells
Patch-Clamp Techniques
levobupivacaine
Colonic Neoplasms
Complementary DNA
Breast Neoplasms

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@article{de774dfe47354bac9f95c6c84465c8aa,
title = "Potent Inactivation-Dependent Inhibition of Adult and Neonatal NaV1.5 Channels by Lidocaine and Levobupivacaine",
abstract = "Background: Cardiotoxic effects of local anesthetics (LAs) involve inhibition of Na V1.5 voltagegated Na + channels. Metastatic breast and colon cancer cells also express Na V1.5, predominantly the neonatal splice variant (nNa V1.5) and their inhibition by LAs reduces invasion and migration. It may be advantageous to target cancer cells while sparing cardiac function through selective blockade of nNa V1.5 and/or by preferentially affecting inactivated Na V1.5, which predominate in cancer cells. We tested the hypotheses that lidocaine and levobupivacaine differentially affect (1) adult (aNa V1.5) and nNa V1.5 and (2) the resting and inactivated states of Na V1.5. Methods: The whole-cell voltage-clamp technique was used to evaluate the actions of lidocaine and levobupivacaine on recombinant Na V1.5 channels expressed in HEK-293 cells. Cells were transiently transfected with cDNAs encoding either aNa V1.5 or nNa V1.5. Voltage protocols were applied to determine depolarizing potentials that either activated or inactivated 50{\%} of maximum conductance (V1/2 activation and V1/2 inactivation, respectively). Results: Lidocaine and levobupivacaine potently inhibited aNa V1.5 (IC 50 mean [SD]: 20 [22] and 1 [0.6] μM, respectively) and nNa V1.5 (IC 50 mean [SD]: 17 [10] and 3 [1.6] μM, respectively) at a holding potential of -80 mV. IC 50s differed significantly between lidocaine and levobupivacaine with no influence of splice variant. Levobupivacaine induced a statistically significant depolarizing shift in the V1/2 activation for aNa V1.5 (mean [SD] from -32 [4.6] mV to -26 [8.1] mV) but had no effect on the voltage dependence of activation of nNa V1.5. Lidocaine had no effect on V1/2 activation of either variant but caused a significantly greater depression of maximum current mediated by nNa V1.5 compared to aNa V1.5. Similar statistically significant shifts in the V1/2 inactivation (approximately -10 mV) occurred for both LAs and Na V1.5 variants. Levobupivacaine (1 μM) caused a significantly greater slowing of recovery from inactivation of both variants than did lidocaine (10 μM). Both LAs caused approximately 50{\%} tonic inhibition of aNa V1.5 or nNa V1.5 when holding at -80 mV. Neither LA caused tonic block at a holding potential of either -90 or -120 mV, voltages at which there was little steady-state inactivation. Higher concentrations of either lidocaine (300 μM) or levobupivacaine (100 μM) caused significantly more tonic block at -120 mV. Conclusions: These data demonstrate that low concentrations of the LAs exhibit inactivation-dependent block of Na V1.5, which may provide a rationale for their use to safely inhibit migration and invasion by metastatic cancer cells without cardiotoxicity.",
author = "Taha Elajnaf and Baptista-Hon, {Daniel T.} and Hales, {Tim G.}",
note = "This study was supported by a BJA/RCoA grant awarded to T.G.H. via the National Institute of Academic Anaesthesia, UK.",
year = "2018",
month = "9",
doi = "10.1213/ANE.0000000000003597",
language = "English",
volume = "127",
pages = "650--660",
journal = "Anesthesia and Analgesia",
issn = "0003-2999",
publisher = "Lippincott, Williams & Wilkins",
number = "3",

}

Potent Inactivation-Dependent Inhibition of Adult and Neonatal NaV1.5 Channels by Lidocaine and Levobupivacaine. / Elajnaf, Taha (Lead / Corresponding author); Baptista-Hon, Daniel T.; Hales, Tim G. (Lead / Corresponding author).

In: Anesthesia and Analgesia, Vol. 127, No. 3, 09.2018, p. 650-660.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Potent Inactivation-Dependent Inhibition of Adult and Neonatal NaV1.5 Channels by Lidocaine and Levobupivacaine

AU - Elajnaf, Taha

AU - Baptista-Hon, Daniel T.

AU - Hales, Tim G.

N1 - This study was supported by a BJA/RCoA grant awarded to T.G.H. via the National Institute of Academic Anaesthesia, UK.

PY - 2018/9

Y1 - 2018/9

N2 - Background: Cardiotoxic effects of local anesthetics (LAs) involve inhibition of Na V1.5 voltagegated Na + channels. Metastatic breast and colon cancer cells also express Na V1.5, predominantly the neonatal splice variant (nNa V1.5) and their inhibition by LAs reduces invasion and migration. It may be advantageous to target cancer cells while sparing cardiac function through selective blockade of nNa V1.5 and/or by preferentially affecting inactivated Na V1.5, which predominate in cancer cells. We tested the hypotheses that lidocaine and levobupivacaine differentially affect (1) adult (aNa V1.5) and nNa V1.5 and (2) the resting and inactivated states of Na V1.5. Methods: The whole-cell voltage-clamp technique was used to evaluate the actions of lidocaine and levobupivacaine on recombinant Na V1.5 channels expressed in HEK-293 cells. Cells were transiently transfected with cDNAs encoding either aNa V1.5 or nNa V1.5. Voltage protocols were applied to determine depolarizing potentials that either activated or inactivated 50% of maximum conductance (V1/2 activation and V1/2 inactivation, respectively). Results: Lidocaine and levobupivacaine potently inhibited aNa V1.5 (IC 50 mean [SD]: 20 [22] and 1 [0.6] μM, respectively) and nNa V1.5 (IC 50 mean [SD]: 17 [10] and 3 [1.6] μM, respectively) at a holding potential of -80 mV. IC 50s differed significantly between lidocaine and levobupivacaine with no influence of splice variant. Levobupivacaine induced a statistically significant depolarizing shift in the V1/2 activation for aNa V1.5 (mean [SD] from -32 [4.6] mV to -26 [8.1] mV) but had no effect on the voltage dependence of activation of nNa V1.5. Lidocaine had no effect on V1/2 activation of either variant but caused a significantly greater depression of maximum current mediated by nNa V1.5 compared to aNa V1.5. Similar statistically significant shifts in the V1/2 inactivation (approximately -10 mV) occurred for both LAs and Na V1.5 variants. Levobupivacaine (1 μM) caused a significantly greater slowing of recovery from inactivation of both variants than did lidocaine (10 μM). Both LAs caused approximately 50% tonic inhibition of aNa V1.5 or nNa V1.5 when holding at -80 mV. Neither LA caused tonic block at a holding potential of either -90 or -120 mV, voltages at which there was little steady-state inactivation. Higher concentrations of either lidocaine (300 μM) or levobupivacaine (100 μM) caused significantly more tonic block at -120 mV. Conclusions: These data demonstrate that low concentrations of the LAs exhibit inactivation-dependent block of Na V1.5, which may provide a rationale for their use to safely inhibit migration and invasion by metastatic cancer cells without cardiotoxicity.

AB - Background: Cardiotoxic effects of local anesthetics (LAs) involve inhibition of Na V1.5 voltagegated Na + channels. Metastatic breast and colon cancer cells also express Na V1.5, predominantly the neonatal splice variant (nNa V1.5) and their inhibition by LAs reduces invasion and migration. It may be advantageous to target cancer cells while sparing cardiac function through selective blockade of nNa V1.5 and/or by preferentially affecting inactivated Na V1.5, which predominate in cancer cells. We tested the hypotheses that lidocaine and levobupivacaine differentially affect (1) adult (aNa V1.5) and nNa V1.5 and (2) the resting and inactivated states of Na V1.5. Methods: The whole-cell voltage-clamp technique was used to evaluate the actions of lidocaine and levobupivacaine on recombinant Na V1.5 channels expressed in HEK-293 cells. Cells were transiently transfected with cDNAs encoding either aNa V1.5 or nNa V1.5. Voltage protocols were applied to determine depolarizing potentials that either activated or inactivated 50% of maximum conductance (V1/2 activation and V1/2 inactivation, respectively). Results: Lidocaine and levobupivacaine potently inhibited aNa V1.5 (IC 50 mean [SD]: 20 [22] and 1 [0.6] μM, respectively) and nNa V1.5 (IC 50 mean [SD]: 17 [10] and 3 [1.6] μM, respectively) at a holding potential of -80 mV. IC 50s differed significantly between lidocaine and levobupivacaine with no influence of splice variant. Levobupivacaine induced a statistically significant depolarizing shift in the V1/2 activation for aNa V1.5 (mean [SD] from -32 [4.6] mV to -26 [8.1] mV) but had no effect on the voltage dependence of activation of nNa V1.5. Lidocaine had no effect on V1/2 activation of either variant but caused a significantly greater depression of maximum current mediated by nNa V1.5 compared to aNa V1.5. Similar statistically significant shifts in the V1/2 inactivation (approximately -10 mV) occurred for both LAs and Na V1.5 variants. Levobupivacaine (1 μM) caused a significantly greater slowing of recovery from inactivation of both variants than did lidocaine (10 μM). Both LAs caused approximately 50% tonic inhibition of aNa V1.5 or nNa V1.5 when holding at -80 mV. Neither LA caused tonic block at a holding potential of either -90 or -120 mV, voltages at which there was little steady-state inactivation. Higher concentrations of either lidocaine (300 μM) or levobupivacaine (100 μM) caused significantly more tonic block at -120 mV. Conclusions: These data demonstrate that low concentrations of the LAs exhibit inactivation-dependent block of Na V1.5, which may provide a rationale for their use to safely inhibit migration and invasion by metastatic cancer cells without cardiotoxicity.

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