Cannabidiol Regulates Gene Expression in Encephalitogenic T cells Using Histone Methylation and noncoding RNA during Experimental Autoimmune Encephalomyelitis

Xiaoming Yang,Marpe Bam,Mitzi Nagarkatti,Prakash S Nagarkatti

doi:10.1038/s41598-019-52362-8

Xiaoming Yang, Marpe Bam + Show 2 more

Open Access

https://doi.org/10.1038/s41598-019-52362-8

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Journal: Scientific Reports	Publication Date: Oct 31, 2019
Citations: 16	License type: open-access

Affiliation: University of South Carolina

Abstract

Cannabidiol (CBD) has been shown by our laboratory to attenuate experimental autoimmune encephalomyelitis (EAE), an animal model of multiple sclerosis (MS). In this study, we used microarray and next generation sequencing (NGS)-based approaches to determine whether CBD would alter genome-wide histone modification and gene expression in MOG sensitized lymphocytes. We compared H3K4me3 and H3K27me3 marks in CD4+ T cells from naïve, EAE and CBD treated EAE mice by ChIP-seq. Although the overall methylation level of these two histone marks did not change significantly, the signal intensity and coverage differed in individual genes, suggesting that CBD may modulate gene expression by altering histone methylation. Further analysis showed that these histone methylation signals were differentially enriched in the binding sites of certain transcription factors, such as ZNF143 and FoxA1, suggesting that these transcription factors may play important roles in CBD mediated immune modulation. Using microarray analysis, we found that the expression pattern of many EAE-induced genes was reversed by CBD treatment which was consistent with its effect on attenuating the clinical symptoms of EAE. A unique finding of this study was that the expression of many miRNAs and lncRNAs was dramatically affected by CBD. In summary, this study demonstrates that CBD suppresses inflammation through multiple mechanisms, from histone methylation to miRNA to lncRNA.

Highlights

(c) Relavite expression level of IL-4, IL-13 and IL-5 determined by real time qPCR (The level in the naïve mice was set as 1)
We investigated whether activation of T cells with myelin oligodendrocyte glycoprotein (MOG) antigen would alter gene expression and histone methylation leading to differentiation of T cells into proinflammatory phenotype and whether CBD treatment would reverse these effects
The histone marks differed significantly in the genes of Th2 related cytokines such as IL-4, IL-5 and IL13 (Fig. 1b). Both marks showed lower levels of coverage in naïve cells when compared to MOG-activated cells, suggesting that histone methylation might be important for the expression of these cytokines in MOG activated T cells

Summary

Introduction

(c) Relavite expression level of IL-4, IL-13 and IL-5 determined by real time qPCR (The level in the naïve mice was set as 1). EAE partially mimics the immunopathological process of MS by immunizing mice with myelin-derived antigens such as myelin basic protein (MBP), proteolipid protein (PLP), myelin associated glycoprotein (MAG) and myelin oligodendrocyte glycoprotein (MOG)[13]. These immunodominant antigenic epitopes trigger acute and chronic autoimmune disease. Our studies have suggested the THC and CBD may modulate histone modification and miRNA expression in immune cells. Our results suggested that histone methylation as well as non coding RNAs may play important roles in inflammatory T cell development and that CBD-mediated immune modulation may result from restoration of such alterations

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