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Research ArticleDrug Discovery and Translational Medicine

Identification of a Human Whole Blood–Based Endothelial Cell Impedance Assay for Assessing Clinical Transient Receptor Potential Vanilloid 4 Target Engagement Ex Vivo

Xiaoping Xu, Navin Goyal, Melissa H. Costell, Theresa Roethke, Christian H. James, Kevin S. Thorneloe, Jaclyn Patterson, Patrick Stoy, Krista Goodman, Dennis L. Sprecher and David J. Behm
Journal of Pharmacology and Experimental Therapeutics March 2021, 376 (3) 436-443; DOI: https://doi.org/10.1124/jpet.120.000307
Xiaoping Xu
GlaxoSmithKline, Collegeville, Pennsylvania
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Navin Goyal
GlaxoSmithKline, Collegeville, Pennsylvania
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Melissa H. Costell
GlaxoSmithKline, Collegeville, Pennsylvania
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Theresa Roethke
GlaxoSmithKline, Collegeville, Pennsylvania
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Christian H. James
GlaxoSmithKline, Collegeville, Pennsylvania
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Kevin S. Thorneloe
GlaxoSmithKline, Collegeville, Pennsylvania
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Jaclyn Patterson
GlaxoSmithKline, Collegeville, Pennsylvania
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Patrick Stoy
GlaxoSmithKline, Collegeville, Pennsylvania
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Krista Goodman
GlaxoSmithKline, Collegeville, Pennsylvania
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Dennis L. Sprecher
GlaxoSmithKline, Collegeville, Pennsylvania
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David J. Behm
GlaxoSmithKline, Collegeville, Pennsylvania
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Abstract

Transient receptor potential vanilloid 4 (TRPV4) channels expressed on pulmonary endothelial cells are activated by elevated pulmonary vascular pressure, resulting in endothelial shape change, pulmonary barrier disruption, and edema. As such, TRPV4 blocker GSK2798745 was recently investigated in phase I/IIa trials to reduce pulmonary edema caused by heart failure (HF). In the absence of a suitable TRPV4 target engagement biomarker, we hypothesized that an ex vivo assay could be used to predict pharmacological activity at the intended site of action (endothelial cells) of subjects. In this assay, the ability of GSK2798745 to block TRPV4 agonist GSK1016790-induced impendence reduction in human umbilical vein endothelial cells (HUVECs) in the presence of human whole blood was assessed. Blood from healthy volunteers drawn 1–12 hours after single or repeated dose of GSK2798745 (5 mg) inhibited GSK1016790-induced impedance reduction by ≥85%. Similarly, blood samples from 16 subjects with HF dosed with GSK2798745 (2.4 mg) inhibited GSK1016790-induced HUVEC impedance reduction by ≥58% 1–24 hours after single dosing and ≥78% 1–24 hours after 7 days of repeated dosing. No inhibition was detected using blood from placebo subjects. Using matched GSK2798745 plasma levels, a pharmacokinetic/pharmacodynamic (PK/PD) relationship was calculated as 2.9 nM IC50, consistent with the 6.5 nM IC50 of GSK2798745 obtained from a rat in vivo PK/PD model of pulmonary edema after correcting for rat-to-human differences. These results indicate that circulating levels of GSK2798745 in the recently completed phase I/IIa trials were sufficient to block TRPV4 in lung vascular endothelial cells to a large extent, supporting this dosing regimen for assessing efficacy in HF.

SIGNIFICANCE STATEMENT In the absence of a suitable target engagement biomarker, we developed an ex vivo assay to predict the pharmacological activity of the transient receptor potential vanilloid 4 (TRPV4) blocker GSK2798745 in healthy volunteers and subjects with heart failure (HF) from phase I/IIa trials. The potency values from the ex vivo assay were consistent with those predicted from a rat in vivo pharmacokinetic/pharmacodynamic model of pulmonary edema, strongly suggesting that circulating levels of GSK2798745 were sufficient to robustly block TRPV4, supporting use of GSK2798745 for assessing efficacy in HF.

Footnotes

    • Received August 28, 2020.
    • Accepted December 23, 2020.
  • This work received no external funding. All authors worked for and owned shares in GlaxoSmithKline during the study.

  • https://doi.org/10.1124/jpet.120.000307.

  • Copyright © 2021 by The American Society for Pharmacology and Experimental Therapeutics
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Journal of Pharmacology and Experimental Therapeutics: 376 (3)
Journal of Pharmacology and Experimental Therapeutics
Vol. 376, Issue 3
1 Mar 2021
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Research ArticleDrug Discovery and Translational Medicine

Cellular Impedance Assay to Predict Human TRPV4 Inhibition

Xiaoping Xu, Navin Goyal, Melissa H. Costell, Theresa Roethke, Christian H. James, Kevin S. Thorneloe, Jaclyn Patterson, Patrick Stoy, Krista Goodman, Dennis L. Sprecher and David J. Behm
Journal of Pharmacology and Experimental Therapeutics March 1, 2021, 376 (3) 436-443; DOI: https://doi.org/10.1124/jpet.120.000307

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Research ArticleDrug Discovery and Translational Medicine

Cellular Impedance Assay to Predict Human TRPV4 Inhibition

Xiaoping Xu, Navin Goyal, Melissa H. Costell, Theresa Roethke, Christian H. James, Kevin S. Thorneloe, Jaclyn Patterson, Patrick Stoy, Krista Goodman, Dennis L. Sprecher and David J. Behm
Journal of Pharmacology and Experimental Therapeutics March 1, 2021, 376 (3) 436-443; DOI: https://doi.org/10.1124/jpet.120.000307
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