A predictive in vitro model of the impact of drugs with anticholinergic properties on human neuronal and astrocytic systems

Elizabeth K. Woehrling, H. Rheinallt Parri, Erin H.Y. Tse, Eric J. Hill, Ian D. Maidment, G. Christopher Fox, Michael D. Coleman

Research output: Contribution to journalArticle

Abstract

The link between off-target anticholinergic effects of medications and acute cognitive impairment in older adults requires urgent investigation. We aimed to determine whether a relevant in vitro model may aid the identification of anticholinergic responses to drugs and the prediction of anticholinergic risk during polypharmacy. In this preliminary study we employed a co-culture of human-derived neurons and astrocytes (NT2.N/A) derived from the NT2 cell line. NT2.N/A cells possess much of the functionality of mature neurons and astrocytes, key cholinergic phenotypic markers and muscarinic acetylcholine receptors (mAChRs). The cholinergic response of NT2 astrocytes to the mAChR agonist oxotremorine was examined using the fluorescent dye fluo-4 to quantitate increases in intracellular calcium [Ca2+]i. Inhibition of this response by drugs classified as severe (dicycloverine, amitriptyline), moderate (cyclobenzaprine) and possible (cimetidine) on the Anticholinergic Cognitive Burden (ACB) scale, was examined after exposure to individual and pairs of compounds. Individually, dicycloverine had the most significant effect regarding inhibition of the astrocytic cholinergic response to oxotremorine, followed by amitriptyline then cyclobenzaprine and cimetidine, in agreement with the ACB scale. In combination, dicycloverine with cyclobenzaprine had the most significant effect, followed by dicycloverine with amitriptyline. The order of potency of the drugs in combination frequently disagreed with predicted ACB scores derived from summation of the individual drug scores, suggesting current scales may underestimate the effect of polypharmacy. Overall, this NT2.N/A model may be appropriate for further investigation of adverse anticholinergic effects of multiple medications, in order to inform clinical choices of suitable drug use in the elderly.

LanguageEnglish
Article numbere0118786
Number of pages18
JournalPLoS ONE
Volume10
Issue number3
DOIs
Publication statusPublished - 4 Mar 2015

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Cholinergic Antagonists
astrocytes
cholinergic agents
cimetidine
drugs
Amitriptyline
Pharmaceutical Preparations
drug therapy
Astrocytes
Cholinergic Agents
Oxotremorine
neurons
Polypharmacy
calcium
Cimetidine
combination drug therapy
cholinergic receptors
fluorescent dyes
coculture
Neurons

Bibliographical note

Copyright: © 2015 Woehrling et al. This is an open access article distributed under the terms of the Creative Commons Attribution License, which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.

Funding: British Brain Research Fund, United Kingdom (United Kingdom Registered Charity: 1140361), Grant Number: 012013

Cite this

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title = "A predictive in vitro model of the impact of drugs with anticholinergic properties on human neuronal and astrocytic systems",
abstract = "The link between off-target anticholinergic effects of medications and acute cognitive impairment in older adults requires urgent investigation. We aimed to determine whether a relevant in vitro model may aid the identification of anticholinergic responses to drugs and the prediction of anticholinergic risk during polypharmacy. In this preliminary study we employed a co-culture of human-derived neurons and astrocytes (NT2.N/A) derived from the NT2 cell line. NT2.N/A cells possess much of the functionality of mature neurons and astrocytes, key cholinergic phenotypic markers and muscarinic acetylcholine receptors (mAChRs). The cholinergic response of NT2 astrocytes to the mAChR agonist oxotremorine was examined using the fluorescent dye fluo-4 to quantitate increases in intracellular calcium [Ca2+]i. Inhibition of this response by drugs classified as severe (dicycloverine, amitriptyline), moderate (cyclobenzaprine) and possible (cimetidine) on the Anticholinergic Cognitive Burden (ACB) scale, was examined after exposure to individual and pairs of compounds. Individually, dicycloverine had the most significant effect regarding inhibition of the astrocytic cholinergic response to oxotremorine, followed by amitriptyline then cyclobenzaprine and cimetidine, in agreement with the ACB scale. In combination, dicycloverine with cyclobenzaprine had the most significant effect, followed by dicycloverine with amitriptyline. The order of potency of the drugs in combination frequently disagreed with predicted ACB scores derived from summation of the individual drug scores, suggesting current scales may underestimate the effect of polypharmacy. Overall, this NT2.N/A model may be appropriate for further investigation of adverse anticholinergic effects of multiple medications, in order to inform clinical choices of suitable drug use in the elderly.",
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A predictive in vitro model of the impact of drugs with anticholinergic properties on human neuronal and astrocytic systems. / Woehrling, Elizabeth K.; Parri, H. Rheinallt; Tse, Erin H.Y.; Hill, Eric J.; Maidment, Ian D.; Fox, G. Christopher; Coleman, Michael D.

In: PLoS ONE, Vol. 10, No. 3, e0118786, 04.03.2015.

Research output: Contribution to journalArticle

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AU - Woehrling, Elizabeth K.

AU - Parri, H. Rheinallt

AU - Tse, Erin H.Y.

AU - Hill, Eric J.

AU - Maidment, Ian D.

AU - Fox, G. Christopher

AU - Coleman, Michael D.

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