Interference on Cytosolic DNA Activation Attenuates Sepsis Severity: Experiments on Cyclic GMP–AMP Synthase (cGAS) Deficient Mice

Although the enhanced responses against serum cell-free DNA (cfDNA) in cases of sepsis—a life-threatening organ dysfunction due to systemic infection—are understood, the influence of the cytosolic DNA receptor cGAS (cyclic guanosine monophosphate–adenosine monophosphate (GMP–AMP) synthase) on sepsis...

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Autores principales:	Peerapat Visitchanakun, Warerat Kaewduangduen, Awirut Chareonsappakit, Paweena Susantitaphong, Prapaporn Pisitkun, Patcharee Ritprajak, Natavudh Townamchai, Asada Leelahavanichkul
Formato:	article
Lenguaje:	EN
Publicado:	MDPI AG 2021
Materias:	sepsis cell free DNA LPS cGAS cecal ligation and puncture Biology (General) QH301-705.5 Chemistry QD1-999
Acceso en línea:	https://doaj.org/article/a876e26c5e8e4df4a241dd4630b39a7f
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spelling	oai:doaj.org-article:a876e26c5e8e4df4a241dd4630b39a7f2021-11-11T16:54:46ZInterference on Cytosolic DNA Activation Attenuates Sepsis Severity: Experiments on Cyclic GMP–AMP Synthase (cGAS) Deficient Mice10.3390/ijms2221114501422-00671661-6596https://doaj.org/article/a876e26c5e8e4df4a241dd4630b39a7f2021-10-01T00:00:00Zhttps://www.mdpi.com/1422-0067/22/21/11450https://doaj.org/toc/1661-6596https://doaj.org/toc/1422-0067Although the enhanced responses against serum cell-free DNA (cfDNA) in cases of sepsis—a life-threatening organ dysfunction due to systemic infection—are understood, the influence of the cytosolic DNA receptor cGAS (cyclic guanosine monophosphate–adenosine monophosphate (GMP–AMP) synthase) on sepsis is still unclear. Here, experiments on cGAS deficient (cGAS<sup>-/-</sup>) mice were conducted using cecal ligation and puncture (CLP) and lipopolysaccharide (LPS) injection sepsis models and macrophages. Severity of CLP in cGAS<sup>-/-</sup> mice was less severe than in wildtype (WT) mice, as indicated by mortality, serum LPS, cfDNA, leukopenia, cytokines (TNF-α, IL-6 and IL-10), organ histology (lung, liver and kidney) and spleen apoptosis. With the LPS injection model, serum cytokines in cGAS<sup>-/-</sup> mice were lower than in WT mice, despite the similar serum cfDNA level. Likewise, in LPS-activated WT macrophages, the expression of several mitochondria-associated genes (as revealed by RNA sequencing analysis) and a profound reduction in mitochondrial parameters, including maximal respiration (determined by extracellular flux analysis), DNA (mtDNA) and mitochondrial abundance (revealed by fluorescent staining), were demonstrated. These data implied the impact of cfDNA resulting from LPS-induced cell injury. In parallel, an additive effect of bacterial DNA on LPS, seen in comparison with LPS alone, was demonstrated in WT macrophages, but not in cGAS<sup>-/-</sup> cells, as indicated by supernatant cytokines (TNF-α and IL-6), M1 proinflammatory polarization (iNOS and IL-1β), cGAS, IFN-γ and supernatant cyclic GMP–AMP (cGAMP). In conclusion, cGAS activation by cfDNA from hosts (especially mtDNA) and bacteria was found to induce an additive proinflammatory effect on LPS-activated macrophages which was perhaps responsible for the more pronounced sepsis hyperinflammation observed in WT mice compared with the cGAS<sup>-/-</sup> group.Peerapat VisitchanakunWarerat KaewduangduenAwirut ChareonsappakitPaweena SusantitaphongPrapaporn PisitkunPatcharee RitprajakNatavudh TownamchaiAsada LeelahavanichkulMDPI AGarticlesepsiscell free DNALPScGAScecal ligation and punctureBiology (General)QH301-705.5ChemistryQD1-999ENInternational Journal of Molecular Sciences, Vol 22, Iss 11450, p 11450 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	sepsis cell free DNA LPS cGAS cecal ligation and puncture Biology (General) QH301-705.5 Chemistry QD1-999
spellingShingle	sepsis cell free DNA LPS cGAS cecal ligation and puncture Biology (General) QH301-705.5 Chemistry QD1-999 Peerapat Visitchanakun Warerat Kaewduangduen Awirut Chareonsappakit Paweena Susantitaphong Prapaporn Pisitkun Patcharee Ritprajak Natavudh Townamchai Asada Leelahavanichkul Interference on Cytosolic DNA Activation Attenuates Sepsis Severity: Experiments on Cyclic GMP–AMP Synthase (cGAS) Deficient Mice
description	Although the enhanced responses against serum cell-free DNA (cfDNA) in cases of sepsis—a life-threatening organ dysfunction due to systemic infection—are understood, the influence of the cytosolic DNA receptor cGAS (cyclic guanosine monophosphate–adenosine monophosphate (GMP–AMP) synthase) on sepsis is still unclear. Here, experiments on cGAS deficient (cGAS<sup>-/-</sup>) mice were conducted using cecal ligation and puncture (CLP) and lipopolysaccharide (LPS) injection sepsis models and macrophages. Severity of CLP in cGAS<sup>-/-</sup> mice was less severe than in wildtype (WT) mice, as indicated by mortality, serum LPS, cfDNA, leukopenia, cytokines (TNF-α, IL-6 and IL-10), organ histology (lung, liver and kidney) and spleen apoptosis. With the LPS injection model, serum cytokines in cGAS<sup>-/-</sup> mice were lower than in WT mice, despite the similar serum cfDNA level. Likewise, in LPS-activated WT macrophages, the expression of several mitochondria-associated genes (as revealed by RNA sequencing analysis) and a profound reduction in mitochondrial parameters, including maximal respiration (determined by extracellular flux analysis), DNA (mtDNA) and mitochondrial abundance (revealed by fluorescent staining), were demonstrated. These data implied the impact of cfDNA resulting from LPS-induced cell injury. In parallel, an additive effect of bacterial DNA on LPS, seen in comparison with LPS alone, was demonstrated in WT macrophages, but not in cGAS<sup>-/-</sup> cells, as indicated by supernatant cytokines (TNF-α and IL-6), M1 proinflammatory polarization (iNOS and IL-1β), cGAS, IFN-γ and supernatant cyclic GMP–AMP (cGAMP). In conclusion, cGAS activation by cfDNA from hosts (especially mtDNA) and bacteria was found to induce an additive proinflammatory effect on LPS-activated macrophages which was perhaps responsible for the more pronounced sepsis hyperinflammation observed in WT mice compared with the cGAS<sup>-/-</sup> group.
format	article
author	Peerapat Visitchanakun Warerat Kaewduangduen Awirut Chareonsappakit Paweena Susantitaphong Prapaporn Pisitkun Patcharee Ritprajak Natavudh Townamchai Asada Leelahavanichkul
author_facet	Peerapat Visitchanakun Warerat Kaewduangduen Awirut Chareonsappakit Paweena Susantitaphong Prapaporn Pisitkun Patcharee Ritprajak Natavudh Townamchai Asada Leelahavanichkul
author_sort	Peerapat Visitchanakun
title	Interference on Cytosolic DNA Activation Attenuates Sepsis Severity: Experiments on Cyclic GMP–AMP Synthase (cGAS) Deficient Mice
title_short	Interference on Cytosolic DNA Activation Attenuates Sepsis Severity: Experiments on Cyclic GMP–AMP Synthase (cGAS) Deficient Mice
title_full	Interference on Cytosolic DNA Activation Attenuates Sepsis Severity: Experiments on Cyclic GMP–AMP Synthase (cGAS) Deficient Mice
title_fullStr	Interference on Cytosolic DNA Activation Attenuates Sepsis Severity: Experiments on Cyclic GMP–AMP Synthase (cGAS) Deficient Mice
title_full_unstemmed	Interference on Cytosolic DNA Activation Attenuates Sepsis Severity: Experiments on Cyclic GMP–AMP Synthase (cGAS) Deficient Mice
title_sort	interference on cytosolic dna activation attenuates sepsis severity: experiments on cyclic gmp–amp synthase (cgas) deficient mice
publisher	MDPI AG
publishDate	2021
url	https://doaj.org/article/a876e26c5e8e4df4a241dd4630b39a7f
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Interference on Cytosolic DNA Activation Attenuates Sepsis Severity: Experiments on Cyclic GMP–AMP Synthase (cGAS) Deficient Mice

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