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Selected TLR7/8 agonist and type I interferon (IFN-α) cooperatively redefine the microglia transcriptome

  • Mst Reshma Khatun
  • Sarder ArifuzzamanEmail author
Original Article

Abstract

Background

Microglia, the primary immune cells of the central nervous system, exerts multiple functions to mediate many neurological diseases. Upon any detection of invading pathogen products (e.g., TLR agonists) or host-released signaling factors (e.g., interferon/IFN), these cells undergo an activation process to release large numbers of inflammatory substances that participate in inflammation and homeostasis. The profound effects of inflammation associated with TLR7/8 agonist Resiquimod (R848) and type 1 interferon (e.g., IFN-α)-induced macrophage and dendritic cell activation on biological outcomes have long been recognized. However, the underlying mechanisms are not well defined in microglial cells.

Methods

The present study investigated the molecular signatures of microglia and identified genes that are uniquely or synergistically expressed in R848-, IFN-α- or R848 with IFN-α-treated primary microglial (PM) cells. We used RNA-sequencing, quantitative real-time PCR, and bioinformatics approaches to derive regulatory networks that control the transcriptional response of PM to R848, IFN-α and R848 with IFN-α.

Results

Our approach revealed that the inflammatory response in R848 with IFN-α-treated PM is faster and more intense than that in R848 or IFN-α-treated PM in terms of the number of differentially expressed genes and the magnitude of induction/repression. In particular, our integrative analysis enabled us to suggest the regulatory functions of TFs, which allowed the construction of a network model that explains how TLR7/8 and IFN-α-sensing pathways achieve specificity.

Conclusion

In conclusion, the systematic approach presented herein could be important to the understanding microglial activation-mediated molecular signatures induced by inflammatory stimuli related to TLR7/8, IFN-α or co-signaling, and associated transcriptional machinery of microglial functions and neuroinflammatory mechanisms.

Keywords

Microglial RNA sequencing Transcription factors Type 1 interferons Toll-like receptor 

Abbreviations

BP

Biological process

CNS

Central nervous system

CSF

Cerebrospinal fluid

cAMP

Cyclic AMP

STAT

Signal transduction and activation of transcription

IRF

Interferon regulatory factors

DAVID

Database for annotation, visualization and integrated discovery

DEG

Differentially expressed gene

DMEM

Dulbecco’s modified Eagle’s medium

EAE

Experimental autoimmune encephalomyelitis

FBP

Fructose-1,6-bisphosphatase

FBS

Fetal bovine serum

GAPDH

Glyceraldehyde-3-phosphate dehydrogenase

GO

Gene ontology

GRO

Growth-regulated oncogenes

HDAC

Histone deacetylase genes

HMOX

Heme oxygenase

IACUC

Institutional animal care and use committee

IFIT

IFN-induced protein with tetratricopeptide

IFN

Interferon

IL

Interleukin

IPA

Ingenuity pathway analysis

ISG

IFN-stimulated gene

KDM

Lysine (K)-specific demethylase

KLF

Kruppel-like factor

R848

Resiquimod

LPS

Lipopolysaccharide

MCP

Monocyte chemoattractant proteins

MCSF

Macrophage colony-stimulating factor

MF

Molecular function

MS

Multiple sclerosis

PCA

Principal component analysis

PC

Principle component

PGE2

Prostaglandin E2

PM

Primary microglia

RNA-seq

RNA sequencing

ROS

Reactive oxygen species

SDF

Stromal-derived factor

TBI

Traumatic brain injury

TF

Transcription factor

TGF-ß

Transforming growth factor beta

TLR

Toll-like receptor

TNF-α

Tumor necrosis factor-alpha

Notes

Acknowledgements

This work was supported by Chung-Ang University Young Scientist Scholarship (CAYSS) program.

Author contributions

MRK and SA conceived the study and interpreted the data.

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interest related to this work.

Ethics approval and consent to participate

All experimental protocols were performed in accordance with Institutional Animal Care and Use Committee (IACUC) guidelines and approved by the IACUC committee of Chung-Ang University.

Supplementary material

10787_2019_610_MOESM1_ESM.tif (6.7 mb)
Supplementary material 1 (TIFF 6863 kb)
10787_2019_610_MOESM2_ESM.tif (3.3 mb)
Supplementary material 2 (TIFF 3427 kb)

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Copyright information

© Springer Nature Switzerland AG 2019

Authors and Affiliations

  1. 1.Department of Biomedical ScienceAjou UniversitySuwonRepublic of Korea
  2. 2.Department of Animal Science and TechnologyChung-Ang UniversityAnseongRepublic of Korea

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