Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells

Thomas Broome, S; Fisher, T; Faiz, A; Keay, KA; Musumeci, G; Al-Badri, G; Castorina, A

Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells

Thomas Broome, S Fisher, T Faiz, A

Keay, KA Musumeci, G Al-Badri, G

Castorina, A

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Publisher:: MDPI
Publication Type:: Journal Article
Citation:: Cells, 2021, 10, (6)
Issue Date:: 2021-05-25

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Field	Value	Language
dc.contributor.author	Thomas Broome, S
dc.contributor.author	Fisher, T
dc.contributor.author	Faiz, A https://orcid.org/0000-0003-1740-3538
dc.contributor.author	Keay, KA
dc.contributor.author	Musumeci, G
dc.contributor.author	Al-Badri, G https://orcid.org/0000-0003-3342-2376
dc.contributor.author	Castorina, A https://orcid.org/0000-0001-7037-759X
dc.date.accessioned	2021-12-01T09:46:55Z
dc.date.available	2021-05-17
dc.date.available	2021-12-01T09:46:55Z
dc.date.issued	2021-05-25
dc.identifier.citation	Cells, 2021, 10, (6)
dc.identifier.issn	2073-4409
dc.identifier.issn	2073-4409
dc.identifier.uri	http://hdl.handle.net/10453/151996
dc.description.abstract	Buspirone is an anxiolytic drug with robust serotonin receptor 1A (Htr1a) agonist activities. However, evidence has demonstrated that this drug also targets the dopamine D3 receptor (Drd3), where it acts as a potent antagonist. In vivo, Drd3 blockade is neuroprotective and reduces inflammation in models of Parkinson's disease. To test if buspirone also elicited anti-inflammatory activities in vitro, we generated stable Drd3<sup>-/-</sup> and Htr1a<sup>-/-</sup> BV2 microglial cell lines using CRISPR-Cas9 technology and then tested the effects of buspirone after lipopolysaccharide (LPS) challenge. We found that LPS exposure had no effect on cell viability, except in Htr1a<sup>-/-</sup> cells, where viability was reduced (<i>p</i> < 0.001). Drug treatment reduced viability in Drd3<sup>-/-</sup> cells, but not in WT or Htr1a<sup>-/-</sup> cells. Buspirone counteracted LPS-induced NO release, NOS2, IL-1β and TNF-α gene expression in WT cells, whereas it exerted limited effects in Drd3<sup>-/-</sup> or Htr1a<sup>-/-</sup> microglia. In summary, our findings indicate that buspirone attenuates microglial polarization after LPS challenge. These results also highlight some major effects of Drd3 or Htr1a genetic ablation on microglial biology, raising important questions on the complex role of neurotransmitters in regulating microglia functions.
dc.format	Electronic
dc.language	eng
dc.publisher	MDPI
dc.relation.ispartof	Cells
dc.relation.isbasedon	10.3390/cells10061312
dc.rights	info:eu-repo/semantics/openAccess
dc.subject.mesh	Animals
dc.subject.mesh	Anti-Inflammatory Agents
dc.subject.mesh	Buspirone
dc.subject.mesh	Cell Line
dc.subject.mesh	Inflammation
dc.subject.mesh	Mice
dc.subject.mesh	Microglia
dc.subject.mesh	Receptor, Serotonin, 5-HT1A
dc.subject.mesh	Receptors, Dopamine D3
dc.title	Assessing the Anti-Inflammatory Activity of the Anxiolytic Drug Buspirone Using CRISPR-Cas9 Gene Editing in LPS-Stimulated BV-2 Microglial Cells
dc.type	Journal Article
utslib.citation.volume	10
utslib.location.activity	Switzerland
pubs.organisational-group	/University of Technology Sydney
pubs.organisational-group	/University of Technology Sydney/Faculty of Science
pubs.organisational-group	/University of Technology Sydney/Faculty of Science/School of Life Sciences
pubs.organisational-group	/University of Technology Sydney/Centre for Health Technologies (CHT)
utslib.copyright.status	open_access	*
dc.date.updated	2021-12-01T09:46:47Z
pubs.issue	6
pubs.publication-status	Published
pubs.volume	10
utslib.citation.issue	6

Abstract:

Buspirone is an anxiolytic drug with robust serotonin receptor 1A (Htr1a) agonist activities. However, evidence has demonstrated that this drug also targets the dopamine D3 receptor (Drd3), where it acts as a potent antagonist. In vivo, Drd3 blockade is neuroprotective and reduces inflammation in models of Parkinson's disease. To test if buspirone also elicited anti-inflammatory activities in vitro, we generated stable Drd3^-/- and Htr1a^-/- BV2 microglial cell lines using CRISPR-Cas9 technology and then tested the effects of buspirone after lipopolysaccharide (LPS) challenge. We found that LPS exposure had no effect on cell viability, except in Htr1a^-/- cells, where viability was reduced (p < 0.001). Drug treatment reduced viability in Drd3^-/- cells, but not in WT or Htr1a^-/- cells. Buspirone counteracted LPS-induced NO release, NOS2, IL-1β and TNF-α gene expression in WT cells, whereas it exerted limited effects in Drd3^-/- or Htr1a^-/- microglia. In summary, our findings indicate that buspirone attenuates microglial polarization after LPS challenge. These results also highlight some major effects of Drd3 or Htr1a genetic ablation on microglial biology, raising important questions on the complex role of neurotransmitters in regulating microglia functions.

Please use this identifier to cite or link to this item:

http://hdl.handle.net/10453/151996