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LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC

BACKGROUND: Dysregulation of transcription and cytokine expression has been implicated in the pathogenesis of a variety inflammatory diseases. The resulting imbalance between inflammatory and resolving transcriptional programs can cause an overabundance of pro-inflammatory, classically activated mac...

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Autores principales: Thomas, Gregory, Frederick, Elizabeth, Thompson, Lisa, Bar-Or, Raphael, Mulugeta, Yetti, Hausburg, Melissa, Roshon, Michael, Mains, Charles, Bar-Or, David
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7702209/
https://www.ncbi.nlm.nih.gov/pubmed/33256749
http://dx.doi.org/10.1186/s12967-020-02626-z
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author Thomas, Gregory
Frederick, Elizabeth
Thompson, Lisa
Bar-Or, Raphael
Mulugeta, Yetti
Hausburg, Melissa
Roshon, Michael
Mains, Charles
Bar-Or, David
author_facet Thomas, Gregory
Frederick, Elizabeth
Thompson, Lisa
Bar-Or, Raphael
Mulugeta, Yetti
Hausburg, Melissa
Roshon, Michael
Mains, Charles
Bar-Or, David
author_sort Thomas, Gregory
collection PubMed
description BACKGROUND: Dysregulation of transcription and cytokine expression has been implicated in the pathogenesis of a variety inflammatory diseases. The resulting imbalance between inflammatory and resolving transcriptional programs can cause an overabundance of pro-inflammatory, classically activated macrophage type 1 (M1) and/or helper T cell type 1 (Th1) products, such as IFNγ, TNFα, IL1-β, and IL12, that prevent immune switching to resolution and healing. The low molecular weight fraction of human serum albumin (LMWF5A) is a novel biologic drug that is currently under clinical investigation for the treatment of osteoarthritis and the hyper-inflammatory response associated with COVID-19. This study aims to elucidate transcriptional mechanisms of action involved with the ability of LMWF5A to reduce pro-inflammatory cytokine release. METHODS: ELISA arrays were used to identify cytokines and chemokines influenced by LMWF5A treatment of LPS-stimulated peripheral blood mononuclear cells (PBMC). The resulting profiles were analyzed by gene enrichment to gain mechanistic insight into the biologic processes and transcription factors (TFs) underlying the identified differentially expressed cytokines. DNA-binding ELISAs, luciferase reporter assays, and TNFα or IL-1β relative potency were then employed to confirm the involvement of enriched pathways and TFs. RESULTS: LMWF5A was found to significantly inhibit a distinct set of pro-inflammatory cytokines (TNFα, IL-1β, IL-12, CXCL9, CXCL10, and CXCL11) associated with pro-inflammatory M1/Th1 immune profiles. Gene enrichment analysis also suggests these cytokines are, in part, regulated by NF-κB and STAT transcription factors. Data from DNA-binding and reporter assays support this with LMWF5A inhibition of STAT1α DNA-binding activity as well as a reduction in overall NF-κB-driven luciferase expression. Experiments using antagonists specific for the immunomodulatory and NF-κB/STAT-repressing transcription factors, peroxisome proliferator-activated receptor (PPAR)γ and aryl hydrocarbon receptor (AhR), indicate these pathways are involved in the LMWF5A mechanisms of action by reducing LMWF5A drug potency as measured by TNFα and IL-1β release. CONCLUSION: In this report, we provide evidence that LMWF5A reduces pro-inflammatory cytokine release by activating the immunoregulatory transcription factors PPARγ and AhR. In addition, our data indicate that LMWF5A suppresses NF-κB and STAT1α pro-inflammatory pathways. This suggests that LMWF5A acts through these mechanisms to decrease pro-inflammatory transcription factor activity and subsequent inflammatory cytokine production.
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spelling pubmed-77022092020-12-01 LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC Thomas, Gregory Frederick, Elizabeth Thompson, Lisa Bar-Or, Raphael Mulugeta, Yetti Hausburg, Melissa Roshon, Michael Mains, Charles Bar-Or, David J Transl Med Research BACKGROUND: Dysregulation of transcription and cytokine expression has been implicated in the pathogenesis of a variety inflammatory diseases. The resulting imbalance between inflammatory and resolving transcriptional programs can cause an overabundance of pro-inflammatory, classically activated macrophage type 1 (M1) and/or helper T cell type 1 (Th1) products, such as IFNγ, TNFα, IL1-β, and IL12, that prevent immune switching to resolution and healing. The low molecular weight fraction of human serum albumin (LMWF5A) is a novel biologic drug that is currently under clinical investigation for the treatment of osteoarthritis and the hyper-inflammatory response associated with COVID-19. This study aims to elucidate transcriptional mechanisms of action involved with the ability of LMWF5A to reduce pro-inflammatory cytokine release. METHODS: ELISA arrays were used to identify cytokines and chemokines influenced by LMWF5A treatment of LPS-stimulated peripheral blood mononuclear cells (PBMC). The resulting profiles were analyzed by gene enrichment to gain mechanistic insight into the biologic processes and transcription factors (TFs) underlying the identified differentially expressed cytokines. DNA-binding ELISAs, luciferase reporter assays, and TNFα or IL-1β relative potency were then employed to confirm the involvement of enriched pathways and TFs. RESULTS: LMWF5A was found to significantly inhibit a distinct set of pro-inflammatory cytokines (TNFα, IL-1β, IL-12, CXCL9, CXCL10, and CXCL11) associated with pro-inflammatory M1/Th1 immune profiles. Gene enrichment analysis also suggests these cytokines are, in part, regulated by NF-κB and STAT transcription factors. Data from DNA-binding and reporter assays support this with LMWF5A inhibition of STAT1α DNA-binding activity as well as a reduction in overall NF-κB-driven luciferase expression. Experiments using antagonists specific for the immunomodulatory and NF-κB/STAT-repressing transcription factors, peroxisome proliferator-activated receptor (PPAR)γ and aryl hydrocarbon receptor (AhR), indicate these pathways are involved in the LMWF5A mechanisms of action by reducing LMWF5A drug potency as measured by TNFα and IL-1β release. CONCLUSION: In this report, we provide evidence that LMWF5A reduces pro-inflammatory cytokine release by activating the immunoregulatory transcription factors PPARγ and AhR. In addition, our data indicate that LMWF5A suppresses NF-κB and STAT1α pro-inflammatory pathways. This suggests that LMWF5A acts through these mechanisms to decrease pro-inflammatory transcription factor activity and subsequent inflammatory cytokine production. BioMed Central 2020-11-30 /pmc/articles/PMC7702209/ /pubmed/33256749 http://dx.doi.org/10.1186/s12967-020-02626-z Text en © The Author(s) 2020 Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated in a credit line to the data.
spellingShingle Research
Thomas, Gregory
Frederick, Elizabeth
Thompson, Lisa
Bar-Or, Raphael
Mulugeta, Yetti
Hausburg, Melissa
Roshon, Michael
Mains, Charles
Bar-Or, David
LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC
title LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC
title_full LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC
title_fullStr LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC
title_full_unstemmed LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC
title_short LMWF5A suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated PBMC
title_sort lmwf5a suppresses cytokine release by modulating select inflammatory transcription factor activity in stimulated pbmc
topic Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7702209/
https://www.ncbi.nlm.nih.gov/pubmed/33256749
http://dx.doi.org/10.1186/s12967-020-02626-z
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