Visualization and modeling of inhibition of IL-1β and TNF-α mRNA transcription at the single-cell level

Abstract IL-1β and TNF-α are canonical immune response mediators that play key regulatory roles in a wide range of inflammatory responses to both chronic and acute conditions. Here we employ an automated microscopy platform for the analysis of messenger RNA (mRNA) expression of IL-1β and TNF-α at th...

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Autores principales:	Daniel Kalb, Huy D. Vo, Samantha Adikari, Elizabeth Hong-Geller, Brian Munsky, James Werner
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/dead43924b9140089bc699de8172f137
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spelling	oai:doaj.org-article:dead43924b9140089bc699de8172f1372021-12-02T14:33:58ZVisualization and modeling of inhibition of IL-1β and TNF-α mRNA transcription at the single-cell level10.1038/s41598-021-92846-02045-2322https://doaj.org/article/dead43924b9140089bc699de8172f1372021-07-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-92846-0https://doaj.org/toc/2045-2322Abstract IL-1β and TNF-α are canonical immune response mediators that play key regulatory roles in a wide range of inflammatory responses to both chronic and acute conditions. Here we employ an automated microscopy platform for the analysis of messenger RNA (mRNA) expression of IL-1β and TNF-α at the single-cell level. The amount of IL-1β and TNF-α mRNA expressed in a human monocytic leukemia cell line (THP-1) is visualized and counted using single-molecule fluorescent in-situ hybridization (smFISH) following exposure of the cells to lipopolysaccharide (LPS), an outer-membrane component of Gram-negative bacteria. We show that the small molecule inhibitors MG132 (a 26S proteasome inhibitor used to block NF-κB signaling) and U0126 (a MAPK Kinase inhibitor used to block CCAAT-enhancer-binding proteins C/EBP) successfully block IL-1β and TNF-α mRNA expression. Based upon this single-cell mRNA expression data, we screened 36 different mathematical models of gene expression, and found two similar models that capture the effects by which the drugs U0126 and MG132 affect the rates at which the genes transition into highly activated states. When their parameters were informed by the action of each drug independently, both models were able to predict the effects of the combined drug treatment. From our data and models, we postulate that IL-1β is activated by both NF-κB and C/EBP, while TNF-α is predominantly activated by NF-κB. Our combined single-cell experimental and modeling efforts show the interconnection between these two genes and demonstrates how the single-cell responses, including the distribution shapes, mean expression, and kinetics of gene expression, change with inhibition.Daniel KalbHuy D. VoSamantha AdikariElizabeth Hong-GellerBrian MunskyJames WernerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-12 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Daniel Kalb Huy D. Vo Samantha Adikari Elizabeth Hong-Geller Brian Munsky James Werner Visualization and modeling of inhibition of IL-1β and TNF-α mRNA transcription at the single-cell level
description	Abstract IL-1β and TNF-α are canonical immune response mediators that play key regulatory roles in a wide range of inflammatory responses to both chronic and acute conditions. Here we employ an automated microscopy platform for the analysis of messenger RNA (mRNA) expression of IL-1β and TNF-α at the single-cell level. The amount of IL-1β and TNF-α mRNA expressed in a human monocytic leukemia cell line (THP-1) is visualized and counted using single-molecule fluorescent in-situ hybridization (smFISH) following exposure of the cells to lipopolysaccharide (LPS), an outer-membrane component of Gram-negative bacteria. We show that the small molecule inhibitors MG132 (a 26S proteasome inhibitor used to block NF-κB signaling) and U0126 (a MAPK Kinase inhibitor used to block CCAAT-enhancer-binding proteins C/EBP) successfully block IL-1β and TNF-α mRNA expression. Based upon this single-cell mRNA expression data, we screened 36 different mathematical models of gene expression, and found two similar models that capture the effects by which the drugs U0126 and MG132 affect the rates at which the genes transition into highly activated states. When their parameters were informed by the action of each drug independently, both models were able to predict the effects of the combined drug treatment. From our data and models, we postulate that IL-1β is activated by both NF-κB and C/EBP, while TNF-α is predominantly activated by NF-κB. Our combined single-cell experimental and modeling efforts show the interconnection between these two genes and demonstrates how the single-cell responses, including the distribution shapes, mean expression, and kinetics of gene expression, change with inhibition.
format	article
author	Daniel Kalb Huy D. Vo Samantha Adikari Elizabeth Hong-Geller Brian Munsky James Werner
author_facet	Daniel Kalb Huy D. Vo Samantha Adikari Elizabeth Hong-Geller Brian Munsky James Werner
author_sort	Daniel Kalb
title	Visualization and modeling of inhibition of IL-1β and TNF-α mRNA transcription at the single-cell level
title_short	Visualization and modeling of inhibition of IL-1β and TNF-α mRNA transcription at the single-cell level
title_full	Visualization and modeling of inhibition of IL-1β and TNF-α mRNA transcription at the single-cell level
title_fullStr	Visualization and modeling of inhibition of IL-1β and TNF-α mRNA transcription at the single-cell level
title_full_unstemmed	Visualization and modeling of inhibition of IL-1β and TNF-α mRNA transcription at the single-cell level
title_sort	visualization and modeling of inhibition of il-1β and tnf-α mrna transcription at the single-cell level
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/dead43924b9140089bc699de8172f137
work_keys_str_mv	AT danielkalb visualizationandmodelingofinhibitionofil1bandtnfamrnatranscriptionatthesinglecelllevel AT huydvo visualizationandmodelingofinhibitionofil1bandtnfamrnatranscriptionatthesinglecelllevel AT samanthaadikari visualizationandmodelingofinhibitionofil1bandtnfamrnatranscriptionatthesinglecelllevel AT elizabethhonggeller visualizationandmodelingofinhibitionofil1bandtnfamrnatranscriptionatthesinglecelllevel AT brianmunsky visualizationandmodelingofinhibitionofil1bandtnfamrnatranscriptionatthesinglecelllevel AT jameswerner visualizationandmodelingofinhibitionofil1bandtnfamrnatranscriptionatthesinglecelllevel
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Visualization and modeling of inhibition of IL-1β and TNF-α mRNA transcription at the single-cell level

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